Herbal Cake-partitioned Moxibustion Research Articles

Herbal cake-partitioned moxibustion regulates M1 and M2 macrophage polarization of colonic mucosa in rats with Crohn's disease

To observe the effect of herbal cake-partitioned moxibustion (Moxi) on the expressions of inflammatory factors and M1/M2 polarization in colonic mucosal macrophages in Crohn's disease (CD) rats, so as to explore its underlying mechanisms in the treatment of CD. Forty male SD rats were randomly divided into normal, model, Moxi and medication groups (n=10). The CD model was established by enema of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) solution (5%TNBS∶50% alcohol=2∶1, 3 mL/kg), once every 7 days, 4 times altogether. For rats of the Moxi group, cake-partitioned moxibustion was given to "Tianshu" (ST25) and "Qihai" (CV6), two moxa-cones for each acupoint every time, once daily for 10 days. For rats of the medication group, intragastric perfusion of mesalazine solution was given twice daily for 10 days. After the treatment, the colonic mucosa tissue was sampled, and the macrophages were isolated, purified and cultured. The pathological changes of colon tissues were observed by H.E. staining. The ultrastructure of colon tissue was observed by transmission electron microscopy. The expression levels of α7nAChR, NF-κB p65 and TNF-α in colon mucosal macrophages were detected by Western blot. The number of M1 and M2 macrophages in colon mucosa was detected by flow cytometry and immunofluorescence assay. Compared with the normal group, the colon tissue of rats presented huge ulceration and inflammatory manifestations, the junction of colon epithelial cells was loose, the structure of organelles was damaged; the expression level of α7nAChR in macrophages of colon mucosa was significantly decreased (P<0.01), while the expression levels of NF-κB p65 and TNF-α, and the number of M1 and M2 macrophages were increased (P<0.01, P<0.05) in the model group. In comparison with the model group, the morphology and structure of colon mucosa tissues of rats in Moxi and medication groups were improved; the expression level of α7nAChR, the number of M2 macrophage in colon mucosa were significantly increased (P<0.01, P<0.05), while the expression levels of NF-κB p65 and TNF-α, and the number of M1 macrophage were significantly decreased (P<0.01, P<0.05) in both the Moxi and medication groups. Herbal cake-partitioned moxibustion may inhibit NF-κB activation by up-regulating the expression level of α7nAChR to promote the polarization of macrophages from M1 to M2 type, and reduce the proportion of M1 macrophages, inhibit the expression of TNF-α in colonic mucosa of CD rats, so as to relieve the intestinal inflammation.

Clinical efficacy of moxibustion for ulcerative colitis and its influence on vitamin D receptor

ObjectiveTo observe the clinical efficacy of herbal cake-partitioned moxibustion for ulcerative colitis (UC) and elucidate its mechanism by targeting the vitamin D receptor (VDR) signaling pathway.MethodsA total of 63 patients with UC were randomly divided into an observation group (30 cases, treated with herbal cake-partitioned moxibustion) and a control group (33 cases, treated with sham herbal cake-partitioned moxibustion). Moxibustion treatment was performed at Qihai (CV6) and bilateral Tianshu (ST25) and Shangjuxu (ST37), 3 times per week for 12 weeks. The total effective rate, visual analog scale (VAS) score for abdominal bloating and pain, and hospital anxiety and depression scale (HADS) score were compared between the two groups. Enzyme-linked immunosorbent assay was used to detect the concentrations of serum C-reactive protein (CRP), 25-hydroxyvitamin D [25(OH)D], and interleukin-12 (IL-12)/interleukin-23 (IL-23) p40. Immunohistochemistry was used to observe the expression levels of VDR and regenerating gene IV (Reg IV) proteins in colonic mucosa. The expression levels of VDR, cytochrome p450 27B1 (CYP27B1), and Reg IV mRNAs were detected by real-time fluorescence quantitive polymerase chain reaction.ResultsAfter treatment, the total effective rate in the observation group was 86.7%, which was significantly higher than 51.5% in the control group (P<0.05). After treatment, the VAS scores for abdominal bloating and pain in the observation group were significantly decreased (P<0.01), as well as the HADS-depression subscale (HADS-D) and HADS-anxiety subscale (HADS) scores (P<0.05), while only the VAS score for abdominal pain in the control group was reduced (P<0.05), and the improvements of the scores in the observation group were more significant than those in the control group (P<0.05). After treatment, the serum CRP concentrations in both groups and the IL-12/IL-23 p40 concentration in the observation group were significantly decreased (P<0.05), and the concentrations in the observation group were lower than those in the control group (P<0.05). The expression levels of VDR protein and mRNA in the colon in both groups were all increased (P<0.01), and the expression levels of Reg IV protein and mRNA and CYP27B1 mRNA were all decreased in the two groups (P<0.05 or P<0.01); the improvements in the observation group were more notable than those in the control group (P<0.05 or P<0.01).ConclusionHerbal cake-partitioned moxibustion can effectively alleviate abdominal pain and diarrhea in patients with UC, improve depression and anxiety disorders, and regulate the expression of related proteins in the VDR signaling pathway. The mechanism may be related to inhibiting intestinal inflammation by reducing the release of the proinflammatory cytokine IL-12/IL-23 p40.

Effects of herbal cake-partitioned moxibustion on plasma trimethylamine, trimethylamine-N-oxide, and flavin-containing monooxygenase 3 in atherosclerotic rabbits

Effects of herbal cake-partitioned moxibustion on plasma trimethylamine, trimethylamine-N-oxide, and flavin-containing monooxygenase 3 in atherosclerotic rabbits

Effects of herbal cake-partitioned moxibustion on the expression of thyroid autophagy-related factors LC3B and Beclin-1 in rats with autoimmune thyroiditis

ObjectiveTo observe the anti-inflammatory effect, as well as the effect on the expression of microtubule-associated protein light chain 3B (LC3B) and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis (EAT).MethodsFemale Sprague-Dawley rats were randomly divided into a normal group and a modeling group. The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction. After the model was prepared, rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group. In the herbal cake-partitioned moxibustion group, moxibustion was alternately applied to two groups of points [Dazhui (GV14)-Mingmen (GV4) and Tiantu (CV22)-Guanyuan (CV4)], and the treatment continued for 30 d. Rats in the normal and model groups were only fixed identically without intervention. Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining; the concentrations of thyroid peroxidase antibodies (TPOAb), thyroglobulin antibodies (TGAb), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay; real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.ResultsThere were massive follicular destruction, lymphocytic infiltration, and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group. Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group. Compared with the normal group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were increased in the model rats (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group (P<0.05). Compared with the model group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were reduced in the herbal cake-partitioned moxibustion group (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group (P<0.05). The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.ConclusionHerbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb. This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.

Influence of herbal cake-partitioned moxibustion on lumbar functions and inflammatory factors in patients with lumbar disc herniation due to kidney deficiency and blood stasis

Influence of herbal cake-partitioned moxibustion on lumbar functions and inflammatory factors in patients with lumbar disc herniation due to kidney deficiency and blood stasis

Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206, AMPK and TSC2 in rats with Crohn disease

Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206, AMPK and TSC2 in rats with Crohn disease

Evaluation of clinical efficacy and mechanisms of Governor Vessel moxibustion therapy on carotid atherosclerosis

To explore the clinical efficacy and possible efficacy mechanisms of Governor Vessel moxibustion therapy in patients with carotid atherosclerosis (CAS). A total of 100 patients with CAS were randomly divided into Governor Vessel moxibustion therapy group (n=51) and control group (n=49). The patients in the Governor Vessel moxibustion therapy group were treated by herbal cake-partitioned moxibustion ［applied on Governor Vessel from Dazhui (GV14) to Yaoshu (GV2)］, once a week, for 8 weeks. Patients in the control group received oral administration of atorvastatin calcium tablets (10 mg) once daily for 8 weeks. Before and after treatment, the patients' constitution quantized score, body mass index (BMI), carotid intima-media thickness (IMT) and laboratory indicators ［including total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), homocysteine (Hcy), fibrinogen (Fbg), and hypersensitive C-reactive protein (hs-CRP)］ were observed, and their clinical efficacy was evaluated. After the treatment, the constitution score and BMI were significantly reduced, and the IMT, TC, TG, LDL, Fbg and hs-CRP decreased significantly in the patients of the two groups as compared with those before treatment (P<0.05); and the constitution score and BMI were obviously lower in the Governor Vessel moxibustion therapy group than in the control group (P<0.05). In the Governor Vessel moxibustion therapy group, the effective rate of constitution score was 88.2% (45/51), the effective rate of weight loss was 54.9% (28/51), while the control group was 61.2% (30/49) and 34.7% (17/49) respectively, those in the Governor Vessel moxibustion therapy group were significantly higher than in the control group (P<0.05). Regarding to the CAS clinical effect, the effective rate was 80.4% (41/51) in the Governor Vessel moxibustion therapy group, and it was 73.5% (36/49) in the control group. There was no significant difference between the two groups in the CAS effective rate (P>0.05). The Governor Vessel moxibustion therapy can effectively treat CAS, and its mechanism may be related to the improvement of risk factors such as high level of TC, TG, LDL, Fbg, and hs-CRP.

Herbal cake-partitioned moxibustion inhibits colonic autophagy in Crohn’s disease via signaling involving distinct classes of phosphatidylinositol 3-kinases

BACKGROUNDAutophagy is an evolutionarily conserved biological process in eukaryotic cells that involves lysosomal-mediated degradation and recycling of related cellular components. Recent studies have shown that autophagy plays an important role in the pathogenesis of Crohn’s disease (CD). Herbal cake-partitioned moxibustion (HM) has been historically practiced to treat CD. However, the mechanism by which HM regulates colonic autophagy in CD remains unclear.AIMTo observe whether HM can alleviate CD by regulating colonic autophagy and to elucidate the underlying mechanism.METHODSRats were randomly divided into a normal control (NC) group, a CD group, an HM group, an insulin + CD (I + CD) group, an insulin + HM (I + HM) group, a rapamycin + CD (RA + CD) group, and a rapamycin + HM (RA + HM) group. 2,4,6-trinitrobenzenesulfonic acid was administered to establish a CD model. The morphology of the colonic mucosa was observed by hematoxylin-eosin staining, and the formation of autophagosomes was observed by electron microscopy. The expression of autophagy marker microtubule-associated protein 1 light chain 3 beta (LC3B) was observed by immunofluorescence staining. Insulin and rapamycin were used to inhibit and activate colonic autophagy, respectively. The mRNA expression levels of phosphatidylinositol 3-kinase class I (PI3KC1), Akt1, LC3B, sequestosome 1 (p62), and mammalian target of rapamycin (mTOR) were evaluated by RT-qPCR. The protein expression levels of interleukin 18 (IL-18), tumor necrosis factor-α (TNF-α), nuclear factor κB/p65 (NF-κB p65), LC3B, p62, coiled-coil myosin-like BCL2-interacting protein (Beclin-1), p-mTOR, PI3KC1, class III phosphatidylinositol 3-kinase (PI3KC3/Vps34), and p-Akt were evaluated by Western blot analysis.RESULTSCompared with the NC group, the CD group showed severe damage to colon tissues and higher expression levels of IL-18 and NF-κB p65 in colon tissues (P < 0.01 for both). Compared with the CD group, the HM group showed significantly lower levels of these proteins (PIL-18 < 0.01 and Pp65 < 0.05). There were no significant differences in the expression of TNF-α protein in colon tissue among the rat groups. Typical autophagic vesicles were found in both the CD and HM groups. The expression of the autophagy proteins LC3B and Beclin-1 was upregulated (P < 0.01 for both) in the colon tissues of rats in the CD group compared with the NC group, while the protein expression of p62 and p-mTOR was downregulated (P < 0.01 for both). However, these expression trends were significantly reversed in the HM group compared with the CD group (PLC3B < 0.01, PBeclin-1 < 0.05, Pp62 < 0.05, and Pm-TOR < 0.05). Compared with those in the RA + CD group, the mRNA expression levels of PI3KC1, Akt1, mTOR, and p62 in the RA + HM group were significantly higher (PPI3KC1 < 0.01 and PAkt1, mTOR, and p62 < 0.05), while those of LC3B were significantly lower (P < 0.05). Compared with the RA + CD group, the RA + HM group exhibited significantly higher PI3KC1, p-Akt1, and p-mTOR protein levels (PPI3KC1 < 0.01, Pp-Akt1 < 0.05, and Pp-mTOR < 0.01), a higher p62 protein level (P = 0.057), and significantly lower LC3B and Vps34 protein levels (P < 0.01 for both) in colon tissue.CONCLUSIONHM can activate PI3KC1/Akt1/mTOR signaling while inhibiting the PI3KC3 (Vps34)-Beclin-1 protein complex in the colon tissues of CD rats, thereby inhibiting overactivated autophagy and thus exerting a therapeutic effect.

Therapeutic observation on herbal cake-partitioned moxibustion for dysmenorrhea due to deficiency cold

To observe the clinical efficacy of herbal cake-partitioned moxibustion for dysmenorrhea due to deficiency cold. A total of 70 patients with dysmenorrhea who met the inclusion criteria were randomized into a mild moxibustion group and a herbal cake-partitioned moxibustion group by the random number table, with 35 cases in each group. Shenque (CV 8), Zhongji (CV 3) and bilateral Zigong (EX-CA 1) were selected for both groups. The treatment continued for 3 menstrual cycles. The visual analog scale (VAS) and COX menstrual symptom scale (CMSS) were scored in both groups before treatment, after treatment and at the end of the 3rd menstrual cycle after treatment. The clinical efficacy was evaluated at the end of the 3rd menstrual cycle after treatment. After treatment, the clinical efficacy of the herbal cake-partitioned moxibustion group had the tendency to be superior to that of the mild moxibustion group, while there was no statistically significant difference in the overall efficacy between the two groups (P>0.05). The VAS and CMSS scores after treatment and at the follow-up were significantly lower than those before treatment in both groups (all P 0.05). The CMSS scores in both groups were significantly lower than those after treatment (both P 0.05). After treatment, the CMSS score in the herbal cake-partitioned moxibustion group was significantly lower than that in the mild moxibustion group (P 0.05). The herbal cake-partitioned moxibustion has the same therapeutic efficacy for dysmenorrhea as the mild moxibustion; the two moxibustion methods can significantly improve the concomitant symptoms of dysmenorrhea, and the herbal cake-partitioned moxibustion is little better.

Effects of different transdermal penetration enhancers applied to herbal cake-partitioned moxibustion on liver lipids, HSL and HMG-CoA reductase in hyperlipidemia rabbits

To observe the effects of laurocapram and borneol as transdermal penetration enhancers applied to herbal cake-partitioned moxibustion on liver lipids, hormone-sensitive lipase (HSL) and hydroxymethylglutaryl CoA (HMG-CoA) reductase in hyperlipidemia rabbits. Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method, with 8 rats in each group. Rabbits in the blank group were fed routinely with a normal diet; rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model. Rabbits in the blank and the model groups were not given any intervention. After the model was prepared successfully, rabbits in the non-transdermal penetration enhancer group received herbal cake-partitioned moxibustion without transdermal penetration enhancers; rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively. After 4 weeks of treatment, the serum was isolated and enzyme-linked immunosorbent assay (ELISA) was applied for the detection of HSL and HMG-CoA reductase. The liver tissues were isolated, and total cholesterol (TC) and triglycerides (TG) were measured by enzymatic methods. One-step method was applied for high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) detection, and transmission turbidimetry was for apolipoprotein A1 (Apo-A1) and apolipoprotein B (Apo-B) detection. The serum concentrations of the drugs in the laurocapram and the borneol groups were significantly higher than those in the non-transdermal penetration enhancer group (both P<0.05); all drug penetrations in the borneol group were significantly higher than those in the laurocapram group (both P<0.05), except for tanshinone IIA. Compared with the non-transdermal penetration enhancer group, the HSL was significantly increased while the HMG-CoA reductase was significantly decreased in the laurocapram and the borneol groups (both P<0.05); between groups, the HSL in the borneol group was significantly higher than that in the laurocapram group (P<0.05). Compared with the blank group, the levels of LDL-C, TG, TC and Apo-B in rabbit liver were significantly increased in the model group (P<0.05); compared with the model group, the levels of LDL-C, TG, TC and Apo-B in the non-transdermal penetration enhancer, the laurocapram, and the borneol groups were significantly decreased (all P<0.05); between groups, the TG and TC in the laurocapram group and the LDL-C, TG, TC and Apo-B in the borneol group were significantly lower than those in the non-transdermal penetration enhancer group (all P<0.05), and the TG, LDL-C and Apo-B in the borneol group were significantly lower than those in the laurocapram group (all P<0.05). Compared with the blank group, the HDL-C and Apo-A1 were significantly decreased in the model group (both P<0.05), while compared with the model group, the HDL-C and Apo-A1 were significantly increased in the non-transdermal penetration enhancer, the laurocapram, and the borneol groups (all P<0.05). Between groups, the Apo-A1 in the laurocapram group, the HDL-C and Apo-A1 in the borneol group were significantly higher than those in the non-transdermal penetration enhancer group (all P<0.05). The application of laurocapram and borneol, as transdermal penetration enhancers, in herbal cake-partitioned moxibustion can promote the penetration of the drugs in the herbal cake, increase the levels of HDL-C and Apo-A1, improve the metabolism of HSL and HMG-CoA reductase, and also simultaneously reduce the levels of TC, TG, LDL-C and Apo-B in the liver. The transdermal penetration enhancement effect of borneol is slightly better than or equivalent to that of laurocapram.

Herbal cake-partitioned moxibustion combined with Rhinocort spray is superior to Rhinocort spray alone in improving symptoms of moderate-severe allergic rhinitis patients

To observe the clinical effect of herbal cake-partitioned moxibustion combined with Rhinocort spray in the treatment of symptoms, sleep quality and daily life quality of patients with moderate-severe allergic rhinitis(AR)．. A total of 67 patients with persistent moderate-severe AR were recruited in the present study and randomized into a treatment group (n＝35) and a control group (n＝32). The patients in the treatment group were treated by herbal cake-partitioned moxibustion (applied to Dazhui [GV14], and bilateral Feishu [BL13], Shenshu [BL23] and Zhiyang [GV9], 3 moxa-cones/time, once every other day) plus Rhinocort spray (once in the morning and evening separately, 256 µg/d), and patients in the control group treated by Rhinocort spray alone (the same to that mentioned above). All the treatments were given for 4 weeks. The therapeutic effect was evaluated with reference to the "Principles and Recommended Schemes for Diagnosis and Treatment of AR" formulated by Otolaryngology Branch of Chinese Medical Association before and after the treatment. The severity of symptoms of AR was assessed by using visual analogue scale (VAS), the life quality of nasal conjunctivitis assessed using "Rhino-conjunctivitis Quality of Life Questionnaire (RQLQ)"， and the sleep quality assessed using "Pittsburgh Sleep Quality Index (PSQI)" before and after the treatment, followed by 3 months' follow-up survey. After the treatment, the VAS and RQLQ scores in both groups and PSQI of the treatment group were significantly decreased in comparison with their own base-line levels of pre-treatment in each group (P<0.05). The RQLQ and PSQI in the treatment group were substantially lower than those in the control group (P<0.05). No marked difference was found between the two groups in VAS(P>0.05). Follow-up survey showed that the VAS and RQLQ scores in both groups and PSQI of the treatment group were still significantly lower than those of pre-treatment in each group(P<0.05), and the VAS, RQLQ and PSQI scores of the treatment group were significantly lower than those of the control group(P<0.05). After 4 weeks' treatment, of the 32 and 35 AR patients in the control and treatment groups, 7 and 16 experienced marked improvement, 16 and 14 were effective, and 9 and 5 failed, with the effective rate being 71.9% and 85.7%, respectively. The effective rate of the treatment group was significantly superior to that of the control group (P<0.05). Herbal cake-partitioned moxibustion combined with Rhinocort spray has a good therapeutic effect in improving symptoms, sleep and quality of life in patients with persistent moderate-severe AR, which is obviously superior to that of Rhinocort spray alone in improving sleep and quality of life.

Effect of herbal cake-partitioned moxibustion on Leptin/JAK2/STAT3 in lipid-lowering pathway of hyperlipidemia rabbits

Objective: To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits, and to explore the possible mechanism. Methods: Forty New-Zealand rabbits were randomly divided into 5 groups using the random number table method, with 8 rats in each group. Rabbits in the blank group were fed routinely with normal diet; rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model. Rabbits in the blank and the model groups were not treated. After the model was prepared, rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer; rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively. After 4 weeks of treatment, serum was collected for enzyme-linked immunosorbent assay (ELISA), and the liver tissues were isolated for immunohistochemistry, quantitative polymerase chain reaction (qPCR) and Western-blotting (WB) detection. Results: Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group (P 0.05). The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin, Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in the model group were significantly lower than those in the blank group (all P<0.05); compared with the model group, the mRNA expressions of leptin, leptin receptor (LR), JAK2 and STAT3 in the non-transdermal absorption enhancer, the laurocapram and the borneol groups were significantly increased (all P<0.05); compared with the non-transdermal absorption enhancer group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the laurocapram and the borneol groups were significantly increased (all P<0.05); compared with the laurocapram group, the mRNA expressions of leptin, LR, JAK2 and STAT3 in the borneol group were significantly increased (P<0.05). The trend of immunohistochemistry and WB detection results was basically consistent with the qPCR assay results. The immunohistochemistry and WB detection results of phosphorylated JAK2 (phospho-JAK2) and phosphorylated STAT3 (phospho-STAT3) were basically consistent with those of JAK2 and STAT3. Conclusion: The molecular expression of Leptin/JAK2/STAT3 pathway in the hyperlipidemia model rabbits was decreased. The molecular expression of Leptin/JAK2/STAT3 pathway was significantly increased after the herbal cake-partitioned moxibustion. The application of laurocapram and borneol, as transdermal absorption enhancers, in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK2/STAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.

Therapeutic observation on acupuncture-moxibustion at different intervals for persistent allergic rhinitis

Objective: To observe the therapeutic effect of acupuncture-moxibustion at different intervals on persistent allergic rhinitis. Methods: A total of 90 patients conforming to the inclusion criteria were randomized into three groups named A, B and C by randomized block method. Patients in all three groups received the same treatment of acupuncture and herbal cake-partitioned moxibustion at the same acupoints, while the treatment frequency was different. Patients in group A received the treatment once a week, group B twice a week and group C three times a week, and all the treatment lasted for 4 weeks. The total nasal symptom score (TNSS), total ocular symptom score (TOSS) and Sino-nasal outcome test-20 (SNOT-20) were evaluated before and after treatment. The self-rating score of symptoms was evaluated during treatment and 2 weeks after treatment. Results: The total effective rate was 80.0% in group A, 93.3% in group B, and 100.0% in group C. The total effective rate in group A was statistically different from that in group B and group C (both P 0.05). After treatment, scores of TNSS, TOSS and SNOT-20 in all three groups dropped significantly, and statistically different from those before treatment (all P 0.05); after 4-week treatment, the score in group A was higher than that in group B and group C, and showed statistical significant (both P 0.05). Conclusion: All three protocols are effective for allergic rhinitis. With the increase of treatment frequency, the therapeutic efficacy with a treatment frequency of twice a week and three times a week is superior to that of once a week. Frequency of three times a week has a better long-term effect than once and twice a week, together with the least fluctuation of symptoms.

Effect of herbal cake-partitioned moxibustion on renal function and expression of renal connective tissue growth factor, integrin-linked kinase and bone morphogenetic protein-7 in chronic renal failure rabbits

To observe the effect of herbal cake-partitioned moxibustion on renal function and expression of connective tissue growth factor (CTGF), integrin-linked kinase (ILK) and bone morphogenetic protein-7 (BMP-7) in chronic renal failure (CRF) rabbits, so as to reveal its mechanisms underlying improvement of CRF. Twenty-four male New Zealand rabbits were randomly divided into control, model, medication and herbal cake-partitioned moxibustion (moxibustion) groups (n=6 rabbits in each group). The CRF model was established by gavage of suspension of Adenine (150 mg·kg-1·d-1) for 21 days. Herbal cake-partitioned moxibustion was applied to "Mingmen"(GV4) and bilateral "Shenshu"(BL23), "Pishu"(BL20) and for 5 moxa-cones every time. Rabbits of the medication group was treated by gavage of Losartan Potassium (2.33 mg·kg-1·d-1). All the treatments were conducted once daily,12 times a course for consecutive 3 courses with a two-day rest after each course of treatment. Serum creatinine (Scr), blood urea nitrogen (BUN) and 24 h urine protein contents were detected by using an automatic biochemical analyzer. The expression levels of CTGF, ILK and BMP-7 proteins and mRNA in the kidney tissue were determined by Western blot and quantitative real time-PCR, separately. Following modeling, Scr and BUN and 24 h urine protein contents were significantly increased in the model group in comparison with the control group (P<0.01). After the intervention, Scr and BUN contents were all significantly decreased in both the moxibustion and medication groups relevant to the model group (P<0.01), suggesting an improvement of the renal function. Compared with the control group, the expression levels of ILK and CTGF mRNAs and proteins were obviously up-regulated (P<0.01), and those of BMP-7 mRNA and protein significantly decreased in the model group (P<0.01). In comparison with the model group, the expression levels of ILK and CTGF mRNAs and proteins were significantly down-regulated in the two treatment groups (P<0.01, P<0.05), and those of BMP-7 mRNA and protein markedly increased in the two treatment groups (P<0.01). In comparison with the medication group, the expression level of ILK protein was significantly up-regulated (P<0.01) and BMP-7 protein was significantly down-regulated (P<0.01) in the moxibustion group. No significant differences were found between the medication and moxibustion groups in down-regulating the levels of Scr, BUN and 24 h urine protein and expression of ILK mRNA, CTGF mRNA and protein and BMP-7 mRNA(P>0.05). Herbal cake-partitioned moxibustion can improve renal function in CRF rabbits, which may be related to its effects in suppressing the expression of ILK and CTGF, and in up-regulating the expression of BMP-7 in the kidney tissue.

Herbal-cake-partitioned moxibustion of "Shenque" (CV8) has a relative specific effect in relieving abdominal pain and in regulating neuroendocrine-immune network in primary dysmenorrhea rats

To observe the effect of herbal-cake-partitioned moxibustion (HCPM) of "Shenque" (CV8) and "Daheng" (SP15) on abdominal pain, plasma β-endorphin (β-EP), uterine prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, as well as splenetic natural killer cell (NK cell) activity in primary dysmenorrhea (PD) rats, so as to explore the specificity of acupoint function and the underlying mechanisms of moxibustion in relieving dysmenorrhea. A total of 40 female rats were randomized into blank control, model, CV8-direct moxibustion, CV8-HCPM and SP15-HCPM groups (n＝8 rats in each). The PD model was established by subcutaneous injection of estradiol benzoate injection (0.2-0.5 mg/rat) for 10 consecutive days and intraperitoneal injection of oxytocin (2 U) 24 h after the last subcutaneous injection. Moxibustion or herbal-cake (composed of Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Paeoniae Rubra, Cortex Cinnamomi, etc.)-partitioned moxibustion was applied to CV8, SP15 or umbilicus respectively for 7 moxa-cones every time, once daily for 10 successive days. The rats of the control and model groups were also restrained as those in the moxibustion groups. The writhing times within 30 minutes was recorded and the contents of plasma β-EP, uterine PGE2 and PGF2α were detected by ELISA, and NK cell activity was detected using MTT. Compared with the control group, the writhing times and the content of PGF2α in the uterus tissue were significantly increased in the model group (P<0.01), while the contents of plasma β-EP, uterine PGE2 and splenetic NK cell activity were significantly decreased (P<0.01). In comparison with the model group, the writhing times and uterine PGF2α content were obviously down-regulated in the SP15-HCPM, CV8-direct moxibustion and CV8-HCPM groups (P<0.05, P<0.01), and the contents of plasma β-EP and uterine PGE2, and splenetic NK cell activity were significantly increased (P<0.05, P<0.01). The therapeutic effects of CV8-HCPM group were significantly superior to those of SP15-HCPM and CV8-direct moxibustion groups in lowering writhing times and PGF2α level, and in up-regulating β-EP, PGE2 (P<0.05, P<0.01). The NK cell activity of CV8-HCPM group was significantly increased compared with that in the SP15-HCPM group(P<0.05). No significant differences were found between the SP15-HCPM and CV8-direct moxibustion groups in the levels of writhing times, plasma β-EP and uterine PGE2, PGF2α contents and splenetic NK cell activity (P>0.05). Moxibustion of both CV8 and SP15 can relieve abdominal pain in PD rats, which may be closely associated with its effect in suppressing PD-induced decrease of plasma β-EP and uterine PGE2 levels and splenetic NK cell activity and increase of uterine PGF2α. The therapeutic effect of CV8-HCPM is obviously better than that of SP15-HCPM and CV8-direct moxibustion.

Regulatory effect of herbal cake-partitioned moxibustion on cholesterol reverse transport nuclear receptor LXRα in rabbits with atherosclerosis

To observe the effect of herbal cake-partitioned moxibustion on liver X receptor alpha (LXRα) in rabbits with atherosclerosis. Thirty-six male New Zealand rabbits were randomly divided into a normal group, a model group, a herbal cake-partitioned moxibustion group and a simvastatin group according to the random number table method, with 9 rabbits in each group. Rabbits in the model group, the herbal cake-partitioned moxibustion group and the simvastatin group were modeled by high fat feeding method which took 12 weeks. After verification of the successful model, rabbits in the normal group were not treated, in the model group were bundled, in the herbal cake-partitioned moxibustion group were treated with herbal cake-partitioned moxibustion, and those in the simvastatin group were treated with simvastatin, all for a total of 4 weeks. At the end of the experiment, the aorta and liver were observed for pathological changes; serum and liver were used to detect lipid levels; Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect LXRα protein and mRNA expression levels, respectively. Compared with the normal group, the structure of aorta was disordered, the wall was rough and thick, the intima was unsmooth, and the vascular smooth muscle cells were arranged closely and disorderly in the model group, which was consistent with the characteristics of the rabbit model of aortic atherosclerosis. Compared with the model group, the aortic structure was clear, the degree of hepatocyte degeneration was reduced, the serum total cholesterol and low-density lipoprotein levels were significantly decreased (all P<0.01), the high-density lipoprotein level was elevated (P<0.01), and the total liver cholesterol was decreased significantly (P<0.01) in the rabbits of the herbal cake-partitioned moxibustion group and the simvastatin group; compared with the model group, the protein (P<0.01 or P<0.05) and mRNA (P<0.01) expressions of rabbit LXRα in the herbal cake-partitioned moxibustion group and the simvastatin group were increased. Herbal cake-partitioned moxibustion can improve the aortic and hepatic lesions, regulate blood lipid and liver lipid levels, increase the expression of liver cholesterol reverse transport nuclear receptor LXRα, promote reverse cholesterol transport in the rabbits with aortic atherosclerosis, therefore produces an antiatherogenic effect.

Herbal cake-partitioned moxibustion improves intestinal epithelial barrier dysfunction by suppressing TNF-α-mediated apoptosis pathway of intestinal epithelium in rats with Crohn's disease

To observe the effect of herbal cake-partitioned moxibustion (Moxi) on tumor necrosis factor (TNF)-α/ TNF receptor 1 (TNFR1)-associated death domain (TRADD) / Fas-associated death domain (FADD) pathway-mediated apoptosis of intestinal epithelial cells in Crohn's disease (CD) rats, so as to explore its underlying mechanisms in the treatment of CD. Forty-eight SD male rats were randomly divided into normal, model, Moxi and medication groups (n＝12 rats in each). The CD model was established by intra-annual perfusion of 2，4，6-trinitrobenzene sulfonic acid (TNBS) solution (TNBS∶50% alcohol＝2∶1, 3 mL/kg), once every 7 days, 4 times altogether. For rats of the Moxi group, moxibustion was given to "Tianshu" (ST25) and "Qihai" (CV6), two moxa-cones every time, once daily for 10 days. For rats of the medication group, intragastric perfusion of mesalazine solution was given twice daily for 10 days. After the treatment, the colonic epithelium tissue was sampled. The epithelial cells were purified and cultured to establish an in vitro intestinal epithelial barrier, and added with TNF-α (a pro-inflammatory factor, 100 ng/mL) in the culture medium for 24 h for making an increased epithelial permeability model. The permeability of intestinal epithelial cell barrier was evaluated by detecting the fluorescence yellow transmittance of the TNF-α-incubated cell medium. Western blot was used to detect the expression levels of TNFR1, TRADD, receptor-interacting protein 1 (RIP1), FADD and zinc finger protein A20 (A20, a ubiquitination enzyme for inhibiting activation of TRADD and RIP1) of the cultured intestinal epithelium cells. The apoptosis of the TNF-α-incubated intestinal epithelial cells was detected by flow cytometry. After modeling and compared with the normal group, the fluorescence yellow transmittance of intestinal epithelia cells, apoptosis rate, and expression levels of TNFR1, TRADD, and RIP1 proteins were significantly increased (P<0.001, P<0.01), and the expression of A20 was significantly decreased (P<0.01) in the model group. In comparison with the model group, the fluorescence yellow transmittance of intestinal epithelial cells, the apoptosis rate and expression levels of TRADD, RIP1 and FADD were remarkably down-regulated (P<0.001, P<0.01), and the expression of A20 was significantly up-regulated (P<0.01) in both the Moxi and medication groups. Herbal cake-partitioned moxibustion may down-regulate the permeability of intestinal epithelial barrier and the apoptosis of intestinal epithelial cells by way of suppressing TNF-α-mediated cellular apoptosis pathway of intestinal epithelium in CD rats.

A Randomized Controlled Clinical Trial of Treatment of Moderate and Severe Persistent Allergic Rhinitis of Spleen Deficiency Pattern by Herbal Cake-partitioned Moxibustion Combined with Rhinocort Nasal Spray

To evaluate the clinical effect of herbal cake-partitioned moxibustion combined with medication in the treatment of moderate and severe persistent allergic rhinitis (PAR) patients of spleen deficiency pattern. Sixty patients with moderate and severe PAR of spleen deficiency pattern were randomized into medication (Rhinocort) group and herbal cake-partitioned moxibustion combined with medication group (combination group, n=30 cases in each). Medicinal cake-partitioned moxibustion was applied to Yintang (GV 29, for about 20 min), Shenque(CV 8, about 50 min), bilateral Zusanli (ST 36, about 40 min)and Hegu(LI 4, about 40 min)once every other day for successive 8 weeks. All the patients received treatment with Rhinocort nasal spray, 2 jets/day for each nostril (64 µg/jet) for 8 weeks. The patients' conditions (symptoms of nasal sufferings, sleeping, sniffle, eye, emotion, etc.) were assessed by visual analogue scale (VAS) and rhino conjunctivitis quality of life questionnaire (RQLQ, 24 items of 7 aspects, 0-6 points/item), respectively. The spleen deficiency syndrome score was determined according to "Guiding Principles for Clinical Research of New Drugs of Chinese Materia Medica" (2002). The therapeutic effect was assessed by referring to "the Diagnostic and Therapeutic Principles and Recommended Schemes for Allergic Rhinitis" (2004). Following the treatment, the scores of VAS, RQLQ, spleen deficiency syndrome were significantly decreased on the 4thand the 8th week of treatment in both groups in comparison with those of their own individual pre-treatment (P<0.05, P<0.01). Four weeks' follow-up survey showed that the VAS score and RQLQ score of the combination group were obviously lower than those of the medication groups (P<0.05), suggesting a better post-effect of moxibustion. Of the two 30 cases in the medication and combination groups, 9 and 9 experienced marked improvement in their symptoms, 16 and 17 were effective, and 5 and 4 ineffective, with the effective rate being 83.3% and 86.7%, respectively. No significant differences were found between the two groups in the VAS and RQLQ scores on the 4th and 8th week during treatment and in the effective rate (P>0.05). Herbal cake-partitioned moxibustion combined with hormone (Rhinocort) nasal spray is effective in relieving symptoms of moderate and severe PAR patients of spleen deficiency syndrome, and has a better post-effect.

Effect of Different Doses of Herbal Cake-partitioned Moxibustion on Histopathological Changes of Colon Tissue in Ulcerative Colitis Rats

To observe the effect of different-doses of herbal cake-partitioned moxibustion (Moxi) on histopathological changes of the damaged colon tissue in rats with ulcerative colitis (UC), so as to select an optimal dosage of Moxi in the treatment of UC. Sixty-three male Sprague Dawley (SD) rats were randomized into 7 groups: normal control, model, medication, and 1, 2, 3 and 4 moxa-cone Moxi (n＝9 rats per group). The UC model was developed by subcutaneous injection of emulsion (1 mL) containing colon mucosa-prepared protein suspension and complete Freund's adjuvant into the toes, groin and back. On the 38th day, enema of 3% formalin and the aforementioned emulsion was used. Herbal-cake (composed of monkshood, cinnamon, etc.) partitioned Moxi with 1 or 2 moxa-cones (about 5 min/cone) was applied to bilateral "Tianshu" (ST 25) once daily or once every other day. The rat's general conditions (diet, movement, response ability, stool, and body weight) were observed, and histopathological changes (adhesion, ulcer formation and inflammation) of colon tissues were examined after hematoxylin-eosin (HE) staining, and scored (histopathological score). Gross score was given according to the severity of adhesion, ulcer formation and inflammation of colonic tissues under stereo microscope. The average optical density (AOD) values of colonic mucins were detected after periodic acid-schiff (PAS) staining, and those of the sulfated mucus content detected after high iron dia-mine-alcian blue (HID-AB) staining. Compared with the normal group, rats in the model group presented loose stool, or with pus and blood, and slowly increased body weight (P<0.01), obvious congestion of colon with ulcer spots or continuous superficial ulcer regions which had irregular glandular cavities, swelling and serious inflammatory infiltration in lamina propria and submucosa, and increased score of colon tissue damage (P<0.01). PAS and HID-AB staining showed a marked decrease of AOD values of colonic mucins and sulfated mucus in the model group relevant to the normal control group (P<0.01), suggesting a reduction of mucus secretion of intestinal glands. Following the intervention, rats in the Moxi groups presented an increase of the body weight, formed feces, and an improvement of the damaged colon tissues as mucosal healing and inflammatory reduction, and a marked decrease of the damage score relevant to the model rats. No significant differences were found in the gross scores among the medication, 1, 2, 3 and 4 moxa-cone groups (P>0.05). The histopathological scores were significantly lower in the 1 and 4 moxa-cone Moxi groups than in the medication group (P<0.05, P<0.01); and significantly lower in the 1, 3, 4 moxa-cone Moxi groups than in the 2 moxa-cone Moxi group (P<0.05, P<0.01). PAS staining showed a significant increase of the AOD values of colonic mucins in the 1, 2, 3 and 4 moxa-cone and medication groups relevant to the model group (P<0.01); and the AOD values of colonic mucins in the 1, 3, 4 moxa-cone Moxi groups were significantly increased than that in the 2 moxa-cone Moxi group (P<0.05, P<0.01). HID-AB staining showed that the AOD values of sulfated mucus content were significantly higher in the 2 and 4 moxa-cone Moxi groups than in the 3 moxa-cone Moxi group (P<0.01). The two-level two-factor factorial analysis showed an interaction existed between the moxa-cone number and Moxi frequency in reducing the gross score and histopathological score and in facilitating colonic mucin and sulfated mucus secretion. The histopathological score of the 4 moxa-cone Moxi group was significantly lower than that of the 2 moxa-cone Moxi group (P<0.05), and the sulfated mucus content was significantly higher in the 4 moxa-cone group than in the 3 moxa-cone group (P<0.01). The effect of Moxi given on alternate days was superior to that of daily Moxi in improving colonic histological damage. Herbal cake-partitioned moxibustion at ST 25 can promote repair of the damaged colonic tissue and secretion of mucin in UC rats. The number of moxa cones and intervention frequency affect the efficacy of Moxi in improving histopathological changes. The Moxi intervention on alternate days and with 2 moxa-cones every time is recommended.

Effect of herbal cake-partitioned moxibustion on MEK1/2 and ERK1/2 expressions of gastric tissues in rats with spleen deficiency syndrome

To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase (MEK1/2) and extracellular regulatory protein kinase (ERK1/2) in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group (group A), a model group (group B), a ranitidine group (group C), and a herbal cake-partitioned moxibustion group (group D) by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4 °C 200% concentrated Da Huang (Radix et Rhizoma Rhei). After successful modeling, the rats in group C were treated with 25 mg/(kg·bw) ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli (ST 36) and Zhongwan (CV 12), for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/(kg·bw) at 8:00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin (HE) staining. The protein expressions of MEK1/2 and ERK1/2 in the gastric tissues were detected by immunohistochemistry. After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK1/2 and ERK1/2 in gastric tissues of the other groups were significantly higher (P<0.01). Compared with group B, the protein expressions of MEK1/2 and ERK1/2 in group C and D were significantly higher (all P<0.01). Compared with group C, the protein expressions of MEK1/2 and ERK1/2 in group D were significantly higher (P<0.01). Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK1/2 and ERK1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.