Intragastric Administration Of Normal Saline Research Articles

Effect of Huangdisan grain on improving cognitive impairment in VD rats and its mechanism in immune inflammatory response.

Vascular dementia (VD) is the second most common type of dementia after Alzheimer's disease (AD). Although the incidence rate is very high, there is no definitive treatment for VD. And it has serious impact on the quality of life of VD patients. In recent years, more and more studies about the clinical efficacy and pharmacological effects of traditional Chinese medicine (TCM) in the treatment of VD have been conducted. And Huangdisan grain has been used to treat VD patients with a good curative effect in clinic. This study was designed to investigate the effect of Huangdisan grain on the inflammatory response and cognitive function of VD rats modeled by bilateral common carotid artery occlusion (BCCAO), that aimed to improve the treatment methods for VD. 8-week-old healthy SPF male Wistar rats (280±20g) were randomly divided into the normal group (Gn, n=10), sham operated group (Gs, n=10), and operated group (Go, n=35). The VD rat models in Go group were established by BCCAO. 8weeks after surgery, the operated rats were screened by the hidden platform trail of Morris Water Maze (MWM), and the rats with cognitive dysfunction were further randomly divided into the impaired group (Gi, n=10) and TCM group (Gm, n=10). The VD rats in Gm group were given the intragastric administration of Huangdisan grain decoction once a day for 8weeks, and the other groups were given intragastric administration of normal saline. Then the cognitive ability of rats in each group was detected by the MWM Test. The lymphocyte subsets in peripheral blood and hippocampus of rats were measured by flow cytometry. The levels of cytokines (IL-1β, IL-2, IL-4, IL-10, TNF-α, INF-γ, MIP-2, COX-2, iNOS) in peripheral blood and hippocampus were measured by ELISA (enzyme linked immunosorbent assay). The number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus was measured by immunofluorescence. Compared with the Gn group, the escape latencies of the Gi group were prolonged (P<0.01), the time spent in the former platform quadrant was shortened (P<0.01), and the number of times of crossing over the former platform location was reduced (P<0.05). But compared with the Gi group, the escape latencies of Gm group were shortened (P<0.01), the time spent in the former platform quadrant was prolonged (P<0.05), and the number of times of crossing over the former platform location was increased (P<0.05). The number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus of VD rats in Gi group was increased (P<0.01) compared with the Gn group. And the proportions of T Cells, CD4+ T Cells, CD8+ T Cells in the hippocampus were increased (P<0.01). The level of pro-inflammatory cytokines in the hippocampus was increased significantly, such as IL-1β (P<0.01), IL-2 (P<0.01), TNF-α (P<0.05), IFN-γ (P<0.01), COX-2 (P<0.01), MIP-2 (P<0.01) and iNOS (P<0.05). And the level of IL-10 (P<0.01), a kind of anti-inflammatory cytokine, was decreased. The proportions of T Cells (P<0.05), CD4+ T Cells (P<0.01) and NK Cells (P<0.05) in the peripheral blood of the VD rats in Gi group were decreased, and the level of IL-1β, IL-2, TNF-α, IFN-γ, COX-2, MIP-2 and iNOS was increased significantly (P<0.01) compared with the Gn group. Meanwhile, the level of IL-4 and IL-10 was decreased (P<0.01). Huangdisan grain could reduce the number of Iba-1+ CD68+ co-positive cells in the CA1 region of hippocampus (P<0.01), decrease the proportions of T Cells, CD4+ T Cells, CD8+ T Cells and the level of IL-1β, MIP-2 in hippocampus (P<0.01) of VD rats. Moreover, it could rise the proportion of NK Cells (P<0.01) and the level of IL-4 (P<0.05), IL-10 (P<0.05), and decrease the level of IL-1β (P<0.01), IL-2 (P<0.05), TNF-α (P<0.01), IFN-γ (P<0.01), COX-2 (P<0.01) and MIP-2 (P<0.01) in peripheral blood of VD rats. This study indicated that Huangdisan grain could decrease the activation of microglia/macrophages, regulate the proportions of lymphocyte subsets and the level of cytokines, which could adjust the immunologic abnormalities of VD rats, and ultimately improve cognitive function.

Heat-inactivated Streptococcus thermophilus MN002 alleviate lipid metabolism of high fat diet-fed induced obese mice through modulating gut microbiota structure and bile acids

To explore the effects of heat-inactivated Streptococcus thermophilus MN-ZLW-002(MN002) on glucose metabolism, lipid metabolism, gut microbiota and bile acids in high-fat diet fed obese mice. Sixty 3-week-old male C57BL/6 mice were randomly divided into control group, high-fat group and intervention group(n=20). After 1 week of adaptive feeding, the control group was fed with normal chow and continued intragastric administration of normal saline for 12 weeks, the high-fat group was fed with high-fat diet and continued intragastric administration of normal saline for 12 weeks, and the intervention group was fed with high-fat diet and continued intragastric of MN002 for 12 weeks. During the experiment, the body weight, food intake, fasting blood glucose content of mice were measured and feces were collected. At the end of the experiment, the oral glucose tolerance of mice was measured and blood, periintestinal fat, peritestosterone fat and perirenal fat samples were collected. The histopathological changes of liver were observed by hematoxylin-eosin staining. Triglyceride, low density lipoprotein, high density lipoprotein and total cholesterol were detected by automatic biochemical analyzer, bile acids content in feces was detected by liquid chromatography-mass spectrometry, gut microbiota structure of mice was analyzed by 16S rDNA sequencing. Compared with high fat group, serum triglyceride, total cholesterol and perirenal fat in intervention group were significantly decreased(P&lt;0.05), the content of fossil cholic acid sulfate in feces was significantly increased, while the content of ursodeoxycholic acid, porcine deoxycholic acid and deoxycholic acid were significantly decreased(P&lt;0.01). Heat inactivation of MN002 could significantly increase the relative abundance of Ruminiclostridium and Alistipes and reduce the relative abundance of Lactobacillus(P&lt;0.01). Heat-inactivated Streptococcus thermophilus MN002 can regulate the gut microbiota structure and bile acid composition and content of high-fat diet fed mice, thereby alleviating the lipid metabolic disorders caused by high-fat diet.

POS-379 TO INVESTIGATE THE EFFECT OF HERBA LYSIMACHIAE IN THE TREATMENT OF OBSTRUCTIVE NEPHROPATHY BASED ON STAT-1/TREM-1 PATHWAY

Obstructive nephropathy is a major cause of chronic renal failure in children and adults.Clinical and animal experiments have found that renal damage still exists after obstruction is relieved, which is mainly related to irreversible fibrosis changes caused by persistent inflammatory infiltration, and there is no specific medicine for western medicine treatment. Herba lysimachiae can detoxify detumescence, diuresis tonglin, has a certain kidney protection. However, there is still a lack of systematic knowledge about the herba lysimachiae components and mechanism of s. chinensis in the treatment of obstructive nephropathy. Twenty SD rats were randomly divided into Sham operation group (Sham), unilateral ureteral obstruction operation group (UUO), Pennyswort intervention Sham operation group (Sham+Lys), herba lysimachiae intervention model group (UUO+ Lys).From the 1st day after surgery, the decoction of herba lysimachiae was given 24g/kg, once a day, for continuous 2w.Or intragastric administration of normal saline at the same time.Routine biochemical kit was used to detect renal function 2 weeks later.Renal tubule dilatation, mesangial hyperplasia and renal interstitial fibrosis were detected by HE, PAS and Masson staining.Immunohistochemistry and immunofluorescence were used to detect the activation of STAT-1/ TreM-1 pathway, macrophage aggregation (CD68+ iNOS+), renal tubular injury markers (KIM-1), and fibrosis related molecules (α-SMA, TGF-beta) in renal tissues. In the model group, the renal tubules were oppressed by urine retention on the obstructed side. Pathological staining showed that the epithelial cells of the renal tubules were necrotic to varying degrees, and the interstitial was infiltrated by acute and chronic inflammatory cells. Some of the necrotic renal tubules were gradually replaced by fibrous scar, and the formation of renal interstitial fibrosis was visible.Biochemical tests showed that the creatinine and renal injury scores were significantly higher than those of the normal group.Immunohistochemistry/immunofluorescence assay showed increased stat-1 /TREM-1 expression.Abnormal aggregation of M1-type macrophages in renal interstitium, accompanied by renal injury and fibrosis, showed increased expressions of KIM-1, α-SMA and TGF-beta (all P<0.05).In contrast, stat-1 /TREM-1 expression was decreased in renal tissue after herba lysimachiae intervention, and the above inflammatory and fibrosis related indicators were improved (P<0.05). Herba lysimachiae can protect renal function and inhibit renal inflammation and fibrosis in UUO rats, and this mechanism is related to the STAT-1/TREM-1 pathway.

Total flavonoids of Drynariae Rhizoma regulates ER-p38 MAPK signaling pathway to improve scopolamine-induced learning and memory impairments in model mice

This study intended to explore the effect and mechanism of total flavonoids of Drynariae Rhizoma in improving scopola-mine-induced learning and memory impairments in model mice. Ninety four-month-old Kunming(KM) mice were randomly divided into six groups. The ones in the model group and blank group were treated with intragastric administration of normal saline, while those in the medication groups separately received the total flavonoids of Drynariae Rhizoma, Kangnaoshuai Capsules, donepezil, as well as total flavonoids of Rhizoma Drynariae plus estrogen receptor(ER) blocker by gavage. The mouse model of learning and memory impairments was established via intraperitoneal injection of scopolamine. Following the measurement of mouse learning and memory abilities in Morris water maze test, the hippocampal ERβ expression was detected by immunohistochemistry, and the expression levels of ERβ and phosphorylated p38(p-p38) in the hippocampus and B-cell lymphoma 2(Bcl-2), Bcl-2-associated death promoter(Bad), and cysteinyl aspartate-specific protease-3(caspase-3) in the apoptotic system were assayed by Western blot. The contents of malondia-ldehyde(MDA), superoxide dismutase(SOD), and nitric oxide(NO) in the hippocampus were then determined using corresponding kits. Compared with the control group, the model group exhibited significantly prolonged incubation period, reduced frequency of cros-sing the platform, shortened residence time in the target quadrant, lowered ERβ, Bcl-2 and SOD activity in the hippocampus, and increased p-p38/p38, Bad, caspase-3, MDA, and NO. Compared with the model group, the total flavonoids of Rhizoma Drynariae increased the expression of ERβ and SOD in the hippocampus, down-regulated the expression of neuronal pro-apoptotic proteins, up-re-gulated the expression of anti-apoptotic proteins, and reduced p-p38/p38, MDA, and NO. The effects of total flavonoids of Drynariae Rhizoma on the above indexes were reversed by ER blocker. It has been proved that the total flavonoids of Drynariae Rhizoma obviously alleviate scopolamine-induced learning and memory impairments in mice, which may be achieved by regulating the neuronal apoptotic system and oxidative stress via the ER-p38 mitogen-activated protein kinase(ER-p38 MAPK) signaling pathway.

RETRACTED: Echinacoside improves diabetic liver injury by regulating the AMPK/SIRT1 signaling pathway in db/db mice

Echinacoside (ECH) is a natural compound extracted from the stem of the Cistanche deserticola plant, has significant biological properties, including antioxidant, anti-inflammatory, neuroprotective, anti-tumor, hepatoprotective, and immunomodulatory properties. In this study, we aimed to explore the protection effects and mechanisms of ECH on diabetic liver injury in db/db mice. Overall, 6-week-old db/db mice (n=20) were randomly allocated to 2 groups: diabetic model group (db/db group, intragastric administration of normal saline, n=10) and ECH-treated group (db/db+ECH group, n=10). Additionally, the normal control group comprised 6-week-old db/m mice (db/m group, normal saline intragastric administration, n=10). ECH was administered once a day for 10weeks. Weight and fasting blood glucose (FBG) were measured biweekly. HE staining and Oil O staining were used to evaluate liver tissue pathological changes and lipid accumulation respectively. Immunofluorescence staining, Western blot and RT-PCR analysis were used to detect the expression of components of the AMPK/SIRT1 signaling axis. The results showed that the administration of echinacoside for 10weeks could significantly improve liver injury and insulin resistance in db/db mice (p<0.01). Also, echinacoside treatment helped to reduce blood lipids and blood glucose (p<0.01). Moreover, ECH actived AMPK/SIRT1 signaling, upregulated peroxisome proliferator-activated receptor gamma co-activator 1 alpha (PGC-1α), proliferator-activated receptor-α (PPARα), carnitine palmitoyl transferase-1A (CPT1A) in db/db mice (p<0.01). The effect of ECH may be elicited by the activation of the liver AMPK/SIRT1 pathway and its downstream factors to improve adiposity, insulin resistance, and dyslipidemia.

Mechanism of Danggui Sini Decoction in treatment of primary dysmenorrhea based on network pharmacology and molecular docking

Network pharmacology, molecular docking and in vivo experiments were used to explore the pharmacodynamic basis and potential mechanism of Danggui Sini Decoction in the treatment of primary dysmenorrhea(PD). The chemical constituents of Danggui(Angelicae Sinensis Radix), Guizhi(Cinnamomi Ramulus), Tongcao(Tetrapanacis Medulla), Baishao(Paeoniae Radix Alba), Xixin(Asari Radix et Rhizoma), Gancao(Glycyrrhizae Radix et Rhizoma), and Dazao(Jujubae Fructus) from Danggui Sini Decoction were retrieved through TCMSP(Traditional Chinese Medicine Systems Pharmacology Database), and the action targets of Danggui Sini Decoction were collected through DrugBank. &quot;Primary dysmenorrhea&quot; and &quot;dysmenorrhea&quot; were used as the key words to search the corresponding targets in the GeneCards, OMIM and TTD databases, and then the intersection targets of Danggui Sini Decoction and the primary dysmenorrhea targets were taken for reverse screening to obtain the corresponding active ingredients. Cytoscape 3.6.1 software was used to construct a traditional Chinese medicine-compound-target-disease network; STRING database was used to build a protein-protein interaction(PPI) network; Gene ontology(GO) function enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were conducted by using DAVID database. The action mechanism of the intersection targets were then predicted, and a histogram chart and bubble chart were drawn for visualization. Then the top five targets in the PPI network were used for docking with the most compounds. In animal experiments, Sprague Dawley(SD) female rats were used to establish a primary dysmenorrhea model by intraperitoneal injection of diethylstilbestrol once a day. A total of 60 SD female rats were randomly divided into 6 groups, namely control group, model group, Danggui Sini Decoction low(1.5 g·kg~(-1)), medium(3.0 g·kg~(-1)), high(6.0 g·kg~(-1)) dose groups, and ibuprofen(20 mg·kg~(-1)) positive control group, with 10 rats in each group. From day 4, except for the control group, rats in the other groups were given intragastric administration of corresponding drugs, and the control group received intragastric administration of normal saline for 7 consecutive days. The number of writhing before and after the administration, the ute-rine contraction inhibition rate and the uterine index after administration were observed, and ELISA assay was used to detect the levels of prostaglandin-endoperoxide synthase 2(PTGS2) and vascular endothelial growth factor A(VEGFA) in the tissues of each group as well as the levels of serum inflammatory factors interleukin 1(IL-1), interleukin 6(IL-6), and tumor necrosis factor-alpha(TNF-α). According to network analysis, 7 Chinese medicines contained 114 active ingredients, 149 targets, and 30 common target genes with PD were obtained. The key targets included VEGFA, IL6, PTGS2, TNF, etc.; GO function enrichment analysis showed a total of 399 terms(P&lt;0.05) were obtained, 353 of which were biological process(BP) terms, 21 were cell composition(CC) terms, and 25 were molecular function(MF) terms. In KEGG pathway enrichment analysis, 14 signaling pathways were obtained, 3 of which were related to inflammation, namely arachidonic acid metabolism, MAPK signaling pathway and NOD-like receptor signaling pathway. The compounds in Danggui Sini Decoction can play a therapeutic role in the treatment of PD by acting on VEGFA, IL-6, PTGS2, TNF and other targets to regulate arachidonic acid and inflammatory signaling pathways.

Study on the attenuation mechanism of aristolochia manshuriensis compatibility with zingiberis rhizoma based on different methods of boiling and blending

Objective Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed. Methods The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method. Results There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ± 0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01). Conclusions The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec. Key words: Caulis aristolochiae manshuriensis; Zingiberis rhizoma; Compatibility (TCD); Decoction preparing methods; Toxicity of Chinese medicine; Acute kidney injury; Rats

Therapeutic effect of irbesartan combined with atorvastatin calcium in the treatment of rats with coronary heart disease.

This study aimed to investigate the therapeutic effect of irbesartan combined with atorvastatin calcium in the treatment of rats with coronary heart disease. One hundred sixty Wistar rats were selected to establish coronary heart disease model. Rats with coronary heart disease were randomly divided into 4 groups: Model, irbesartan, atorvastatin calcium and combination groups (irbesartan combined with atorvastatin calcium group). Rats in irbesartan group were treated with 50 mg/(kg.day) irbesartan; rats in atorvastatin calcium group were given atorvastatin calcium at a dose of 10 mg/(kg.day); rats in combination group were subjected to atorvastatin calcium at a dose of 10 mg/(kg.day) and irbesartan at a dose of 50 mg/(kg.day), while rats in model groups were given intragastric administration of normal saline at a dose of 2 ml/day. Serum lipids, including total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and TC/HDL-C, were measured by automatic biochemical analyzer. Expression of sPLA2-V in myocardium and aortic trunk of rats was detected by reverse transcription-PCR (RT-PCR) and western blot analysis. After treatment, levels of serum TC, TG, LDL-C, HDL-C and TC/HDL-C in rats of each treatment group were better than those in model group (p<0.05). Expression level of sPLA2-V in myocardium and aortic trunk in model group was significantly higher than that in other groups (p<0.05). Expression level of sPLA2-V in combination group was significantly lower than that in irbesartan and atorvastatin calcium groups (p<0.05). Combination of irbesartan and atorvastatin calcium is superior to irbesartan or atorvastatin calcium alone in the treatment of rats with coronary heart disease. The possible explanation is that the two drugs can reduce the expression of sPLA2-V in myocardium and aortic trunk, which in turn relieved atherosclerosis and achieved better therapeutic effect.

The initial mechanism investigation in immune escape correlated T cell proportion of Lewis lung cancer by Fuzheng-Peiyuan formula

Objective To observe the initial mechanism investigation in immune escaping correlated T cell proportion of Lewis lung cancer by Fuzheng-Peiyuan formula. To provide a theoretical basis for the application of the Chinese medicine of strengthening the body resistance in clinics. Methods The healthy C57BL/6J mice were divided into six groups by the random number table: the Chinese medicine group, the Chinese medicine plus chemotherapy group, the chemotherapy group, the model group and the normal control group. There were 6 mice in each group. All the groups expect the normal control group were inoculated subcutaneously with Lewis lung cancer cell suspension 0.2 ml. The medicine given to the Chinese medicine group were orally Fuzheng-Peiyuan formula with 8.17 g/kg. The combination group was given intraperitoneal injection of cyclophosphamide 60 mg/kg and orally Fuzheng-Peiyuan formula 8.17 g/kg. The chemotherapy group received the injection of cyclophosphamide 60 mg/kg and intragastric administration of normal saline. The model group and normal control group were administered with saline. After continuous administration for 13 days, the expressions of CD4, Foxp3, CD8 and CD28 in spleen cells of tumor bearing mice were observed by flow cytometry. Results Compared with the model group, the chemotherapy group and the chemotherapy combined with Chinese traditional medicine group showed that tumor weight (1.76 ± 0.42 g, 1.40 ± 0.43 g vs. 4.37 ± 0.59 g) significantly decreased (P<0.01); and the CD4+ Foxp3+ cells of mouse spleen cells in the Chinese medicine group and Chinese medicine plus chemotherapy group (11.25% ± 1.69%, 9.30% ± 2.68% vs. 14.21% ± 1.50 %) significantly decreased (P<0.05 or P<0.01). Conclusions The result initial proved that the Chinese medicine could strengthen the body resistance, adjust the proportion of Treg and CTL in spleen T cell in tumor-bearing mouse. Key words: T-lymphocyte subsets; Carcinoma, Lewis lung; Fuzheng-Peiyuan formula; Mice

Effects of telmisartan on improving leptin resistance and inhibiting hepatic fibrosis in rats with non-alcoholic fatty liver disease.

The present study aimed to investigate the impacts of telmisartan (TEL) on hepatic fibrosis, serum leptin, leptin protein in liver tissue and its mRNA expression level in rats with non-alcoholic fatty liver disease (NAFLD). Male Sprague Dawley rats were randomly divided into the control (N), model (M), polyene phosphatidylcholine (P) and TEL (T) groups. Group M and the intervention groups were given a high-fat diet for 12 weeks to induce NAFLD, followed by 4 weeks of intragastric administration of normal saline (1.0 ml/kg/day), polyene phosphatidylcholine (PPC; 123.1 mg/kg/day) and TEL (8 mg/kg/day). The liver tissue was then assessed for the NAFLD activity score and fibrosis score (FS), and serum biochemistry and leptin levels were determined. Additionally, leptin protein expression levels were examined by western blotting and the expression of leptin mRNA was investigated by reverse transcription-polymerase chain reaction. TEL significantly improved FS in rats (P<0.01) and was more effective than PPC. TEL significantly reduced the expression of serum leptin, as well as the expression levels of leptin protein and its mRNA in liver tissue (P<0.01); however, the effects of PPC were not significant (P>0.05). TEL reduced serum leptin, leptin protein and its mRNA in the liver tissue of NAFLD rats, and improved the pathological indicators of liver fibrosis.

Metformin improves cardiac function in mice with heart failure after myocardial infarction by regulating mitochondrial energy metabolism

To investigate whether metformin can improve the cardiac function through improving the mitochondrial function in model of heart failure after myocardial infarction. Male C57/BL6 mice aged about 8 weeks were selected and the anterior descending branch was ligatured to establish the heart failure model after myocardial infarction. The cardiac function was evaluated via ultrasound after 3 days to determine the modeling was successful, and the mice were randomly divided into two groups. Saline group (Saline) received the intragastric administration of normal saline for 4 weeks, and metformin group (Met) received the intragastric administration of metformin for 4 weeks. At the same time, Shame group (Sham) was set up. Changes in cardiac function in mice were detected at 4 weeks after operation. Hearts were taken from mice after 4 weeks, and cell apoptosis in myocardial tissue was detected using TUNEL method; fresh mitochondria were taken and changes in oxygen consumption rate (OCR) and respiratory control rate (RCR) of mitochondria in each group were detected using bio-energy metabolism tester, and change in mitochondrial membrane potential (MMP) of myocardial tissue was detected via JC-1 staining; the expressions and changes in Bcl-2, Bax, Sirt3, PGC-1α and acetylated PGC-1α in myocardial tissue were detected by Western blot. RT-PCR was used to detect mRNA levels in Sirt3 in myocardial tissues. Metformin improved the systolic function of heart failure model rats after myocardial infarction and reduced the apoptosis of myocardial cells after myocardial infarction. Myocardial mitochondrial respiratory function and membrane potential were decreased after myocardial infarction, and metformin treatment significantly improved the mitochondrial respiratory function and mitochondrial membrane potential; Metformin up-regulated the expression of Sirt3 and the activity of PGC-1α in myocardial tissue of heart failure after myocardial infarction. Metformin decreases the acetylation level of PGC-1α through up-regulating Sirt3, mitigates the damage to mitochondrial membrane potential of model of heart failure after myocardial infarction and improves the respiratory function of mitochondria, thus improving the cardiac function of mice.

Icariin Attenuates High-cholesterol Diet Induced Atherosclerosis in Rats by Inhibition of Inflammatory Response and p38 MAPK Signaling Pathway.

Icariin is a flavonoid isolated from the traditional Chinese herbal medicine Epimedium brevicornum Maxim and has been reported to be effective for the treatment of a variety of cardiovascular diseases. The aim of the present study was to investigate the effect and mechanism of icariin on atherosclerosis (AS) using a high-cholesterol diet (HCD)-induced rat model. Seventy male Wistar rats were divided into five groups: 20 in the control group, 20 in the AS group, 10 in the simvastatin group, 10 in the low-dose icariin group, and 10 in the high-dose icariin group. A HCD and vitamin D3 were administered to establish AS rat model. The five groups of rats received daily intragastric administration of normal saline, simvastatin, or icariin (30mg/kg/d, 60mg/kg/d) for 4weeks. The levels of blood lipids, superoxide dismutase (SOD), and malonaldehyde (MDA) were measured. The mRNA levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α were analyzed by real-time RT-PCR, and the serum levels of IL-6 and TNF-α were measured using ELISA kit. In addition, the expression of phosphorylated p38 (p-p38) MAPK was detected by Western blot analysis. The results indicated that AS rat models were successfully constructed. In the AS group, the levels of blood lipids including total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C), and MDA were significantly increased, while high-density lipoprotein-cholesterol (HDL-C) and SOD were significantly decreased, compared with those in the control group. However, icariin succeeded in improving these biochemical parameters towards the normal values in the control group. In the simvastatin group and the icariin groups, the serum levels of IL-6 and TNF-α and the related tissue mRNA levels, as well as the expression of p-p38 MAPK, were markedly reduced compared with the AS group. In conclusion, the present study indicated that icariin inhibited the HCD-induced dyslipidemia in rats, the mechanisms may be associated with the anti-inflammation, anti-oxidative stress, and downregulation of p-p38 MAPK by icariin.

Determination of urine oxalate level in rats with renal calcium oxalate calculus by high-performance liquid chromatography.

To establish a method of high-performance liquid chromatography (HPLC) for determining the urine oxalate levle in rats with renal calcium oxalate calculus. Totally 24 SPF Wistar healthy male rats were randomly divided into control group(n=12)and ethylene glycol (EG) group (n=12). Rats in EG group were administered intragastrically with 2% ammonium chloride (AC)2 ml/rat per day+1% ethylene glycol (EG), along with free access to drinking water.The control group was fed with deionized water, along with the intragastric administration of normal saline (1 ml per day). Twenty-eight days after modelling, the 24-hour urine samples were collected, and the urine oxalic acid levels were determined using HPLC and the results were compared with those of catalytic spectrophotometry using oxidation of methyl. During the HPLC, the samples were separated on Aglient 5TC-C18 (250×4.6 mm,5 Μm), eluted with mixture of methanol (0.1 mol/L) and ammonium acetate (15:85) at 1.2 ml/min, and detected at 314 nm, with the column temperature being 20 ℃. The standard curves of high and low concentrations of oxalic acid were y=5909.1x+378730, R² =0.9984 and y=7810.5x-16635, R² =0.9967,respectively. The lowest detectable concentration in this method was 5 Μg/ml. The linear high concentration range of oxalate stood at 62.50-2000.00 Μg/ml, and the linear low concentration range of oxalate stood at 6.25-100.00 Μg/ml. Its average recovery was 95.1%, and its within-day and day-to-day precisions were 3.4%-10.8% and 3.8%-9.4%. Both HPLC and catalytic spectrophotometry showed significantly higher urinary oxalic acid concentration and 24 h urine oxalate level in EG group compared with the control group [urinary oxalic acid concentration: (736.35 ± 254.52) Μg/ml vs.(51.56 ± 36.34) Μg/ml,(687.35 ± 234.53) Μg/ml vs.(50.24 ± 42.34) Μg/ml;24 h urine oxalate level: (11.23 ± 4.12)mg vs.(0.87 ± 0.45)mg,(9.89 ± 3.55)mg vs. (0.77 ± 0.65)mg; all P<0.01]. No statistically significant difference was observed in the results of urinary oxalate concentration and 24 h urine oxalate level between HPLC and potassium chromate oxidation of methyl red spectrophotometry (all P>0.05). HPLC is a simple, rapid, and precise method in detecting urine oxalate level in rats with renal calcium oxalate calculus, with high recovery rate.

Rosuvastatin Attenuates Atrial Structural Remodelling in Rats with Myocardial Infarction through the Inhibition of the p38 MAPK Signalling Pathway

The purpose of this study was to verify the hypothesis that rosuvastatin attenuates atrial structural remodelling in rats with myocardial infarction (MI) through the regulation of the p38 mitogen-activated protein kinase (MAPK) signalling pathway. A total of 66 rats were used in this study to establish a model of MI. The 56 rats that survived the first 24h after surgery were randomly divided into four groups: the control group (C group), the rosuvastatin group (R group), the low-dose torasemide group (T1 group), and the high-dose torasemide group (T2 group). The four groups of rats received daily intragastric administration of normal saline, rosuvastatin, or torasemide (T1: 1mg/kg body weight; T2: 2mg/kg body weight) for a total of four weeks. The rats in the sham-operated group (n=14) also received daily intragastric administration of normal saline for four weeks. After four weeks of intervention, the left ventricular end-diastolic pressure (LVEDP) was measured in all groups of rats by haemodynamic methods. The rats were then sacrificed, and the left atrial tissues were collected. The collagen volume fractions (CVFs) in the left atrial tissues were determined using Masson's trichrome staining. The expression of phosphorylated p38 (P-p38) MAPK in the left atrial tissues was examined by immunohistochemistry and western blot analysis. The results showed that LVEDP, CVF, and P-p38 MAPK expression were drastically elevated in the four MI groups in comparison to the sham-operated group (p<0.001). Rosuvastatin elevated the left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF). Both rosuvastatin and torasemide improved the haemodynamic parameters. No significant difference was detected in LVEDP between the R group and the T1 group (p=0.37). In contrast, LVEDP was significantly higher in the R group than in the T2 group (p <0.05). CVF (%) was markedly decreased in the R group compared to the C, T1, and T2 groups (decreased by 47.4%, 28%, and 20.1%, respectively). Immunohistochemical analysis showed that the indices of P-p38 MAPK positive cells were significantly decreased in the R group in comparison with the C, T1, and T2 groups (decreased by 44.6%, 36.6%, and 21.4%, respectively). Western blot analysis demonstrated that P-p38 MAPK expression was markedly reduced in the R group compared with the C and T1 groups (reduced by 67% and 40.5%, respectively). The level of P-p38 MAPK in the R group was slightly higher than in the T2 group. However, the difference was not statistically significant (p>0.05). Rosuvastatin attenuates atrial structural remodelling in rats with MI. The mechanism underlying this phenomenon may be associated with the downregulation of P-p38 MAPK by rosuvastatin.

Effects of Ciji Hua'ai Baosheng granule formula (CHBGF) on life time, pathology, peripheral blood cells of tumor chemotherapy model mouse with H22 hepatoma carcinoma cells.

Ciji Hua'ai Baosheng Granule Formula (CHBGF) is a traditional Chinese empirical formula that can help the tumor patients who have received chemotherapy antagonize the toxin and side-effects so as to improve and prolong the life. This study is to evaluate the effects of CHBGF on improving life quality in terms of survival time, pathology of tumor tissue and ameliorating peripheral blood cells in mouse chemotherapy model with subcutaneous transplanted tumor or ascitic tumor of H22 hepatoma carcinoma cells at an overall level. 71 mice among the 92 Kunming mice were injected subcutaneously into the right anterior armpit with H22 hepatoma carcinoma cells, after 7 days, which had formed tumors and were used peritoneal injection of Cytoxan (CTX) (200mg/kg) to establish the mouse chemotherapy model with transplanted tumor, and then which were commensurately divided into 8 groups by random digits table. 21 mice were injected into peritoneal cavity to use CTX and the same method to establish the model. The groups for evaluating the effects on the survival time were the model, CHBGF and positive control group respectively with 7 mice in each group. The groups for evaluating the effects on anti-cancer were the model group, three treatment groups and positive control group with 10 mice in each group. The survival-time-observing groups were given intragastric administration of normal saline, CHBGF (64g/kg) once a day, and peritoneal injection of 5-Fluorouracil (25mg/kg) once every other day respectively. The survival time of each group was observed. The five anti-cancer-observing groups were given intragastric administration of normal saline, CHBGF (64g/kg, 32g/kg and 16g/kg) once a day, and peritoneal injection of 5-Fluorouracil (25mg/kg) once every other day respectively. After treatment for 21 days, the transplanted tumors were peeled off. Blood was collected through pricking eyeball and analyzed by hematology analyzer. And postchemotherapy transplanted tumor inhibition ratios were calculated. Pathological changes of tumor tissues and blood smears were observed with light microscope. The life prolonging rate of CHBGF (64g/kg) group with transplanted tumor is 20.14%, and their survival time was longer than that of the 5-Fluorouracil group (P<0.05). Life prolonging rate of CHBGF (64g/kg) group with ascitic tumor is 64.15%, the survival time was longer than that of the model group (P<0.01) and the 5-Fluorouracil group (P<0.05). The growth of the transplanted tumor in model group was faster than that in CHBGF (64g/kg) group and 5-Fluorouracil group (P<0.05). The tumor average weight of the positive drug and the CHBGF (64g/kg, 32g/kg) groups was lighter than that of the model group (P<0.05 or P<0.01). The inhibition ratios of CHBGF (64g/kg, 32g/kg and 16g/kg) groups are 31.15%, 21.31%, and 13.11% respectively. Under light microscope, in the positive drug and three CHBGF groups the pathological deteriorated severity of tumor tissue observed was milder than that in the model group, the distribution of WBC in CHBGF groups was more obvious than that of the model and 5-Fluorouracil groups. The WBC and PLT decrease in CHBGF (64g/kg, 32g/kg and 16g/kg) groups is less than the model and the 5-Fluorouracil group (P<0.05 or P<0.01), the number of RBC and HGB just in the CHBGF (64g/kg) group was more than that of the model group or the 5-Fluorouracil group (P<0.05). Ciji Hua'ai Baosheng Granule Formula can prolong the survival time of the mice chemotherapy model of both subcutaneous transplanted tumor and ascitic tumor of H22 hepatoma carcinoma cells, has some determinate inhibitory effects on the growth of subcutaneous transplanted tumor chemo-treated, and has the therapeutic effect on antagonizing decrease of WBC and PLT caused by chemotherapy.

Protection effect of atorvastatin in cerebral ischemia-reperfusion injury rats by blocking the mitochondrial permeability transition pore.

The aim of this study was to investigate the influence of atorvastatin on the opening of the mitochondrial permeability transition pore (MPTP) and the expression of cytochrome C (Cyt C) in Sprague-Dawley rats with cerebral ischemia-reperfusion (I/R). The rat model of cerebral artery ischemia was established by the suture-occluded method with ischemia for 2 h and reperfusion for 72 h. Thirty-four male rats were randomly divided into four groups: the normal group and the sham-operation group without any treatment, the I/R group with only intragastric administration of normal saline, and the intervention group, which received intragastric administration of 10 mg/kg atorvastatin at different times. All rats were sacrificed at 72 h. Compared with the I/R group, the morphology of nerve cells in the intervention group was reduced, the number of TUNEL-positive cells decreased, the expression of cortical cytoplasm Cyt C decreased, and the mitochondrial absorbance value increased. All of these differences were statistically significant. Atorvastatin could inhibit neuronal apoptosis and alleviate the cerebral I/R injury. The mechanism may be related to the blocking of the MPTP opening and the subsequent reduction of Cyt C release.

ATTENUATION OF NEOINTIMAL HYPERPLASIA THROUGH DOWN-REGULATION OF INTERLEUKIN-18 EXPRESSION BY CURCUMIN IN RAT BALLOON INJURY MODEL

ObjectivesVascular inflammation could control neointimal hyperplasia processes in vascular healing after injuried. Studies had indicated drug-eluting stents (DES) improved the efficacy of percutaneous coronary intervention by modulating vascular inflammation and...

Effect of Chaihu Shugan Tang on excitability in different brain regions ofpentylenetetrazole-kindled chronic epileptic rats

To study the effect of the Chinese compound prescription Chaihu Shugan Tang (CHSGT) on the excitability in the cerebral cortex and hippocampus (different brain regions) of pentetrazole (PTZ)-kindled chronic epileptic rats. To establish the model of chronic kindling rats intraperitoneal injected with pentylenetet. Fully kindled rats were randomized into control and experimental groups for intragastric administration of normal saline (control, model), Sodium Valproate and CHSGT at the high, medium and low doses for 4 consecutive weeks. The content of 2-NBDG, the glutamate (Glu) and the aspartate (Asp) in different brain regions of rats were detected by fluorescence imaging techniques and HPLC assay respectively. CHSGT at the high, medium and low doses all significantly decreased the content of 2-NBDG, the Glu and the Asp in different brain regions of chronic epileptic rats (P < 0.01). CHSGT can inhibit the excitability in different brain regions of PTZ-induced epileptic rats, by decreasing the level of excitatory neurotransmitter maybe one of its antiepileptic mechanisms.

Effects of Yiqi Huayu Decoction on brain-derived neurotrophic factor expression in rats with lumbar nerve root injury

To observe the effects of Yiqi Huayu Decoction, a compound traditional Chinese herbal medicine, on brain-derived neurotrophic factor (BDNF) expression in lumbar nerve root and soleus of rats after lumbar nerve root compression injury. A total of 120 male SD rats were randomly allocated into sham-operated group, untreated group, methycobal injection group and Yiqi Huayu Decoction group. Surgery were performed on rats of untreated group, methycobal injection group and Yiqi Huayu Decoction group by placing a micro-silica gel on right L(4) dorsal root ganglia, while rats in sham-operated group received skin and paravertebral muscle incision only. Rats in the methycobal injection group and the Yiqi Huayu Decoction group were given intramuscular injection of methycobal and intragastric administration of Yiqi Huayu Decoction respectively. Rats in the sham-operated group and the untreated group received intragastric administration of normal saline of equal volume as Yiqi Huayu Decoction. The compressed nerve roots and right soleus were harvested after 10-, 30- and 60-day treatment. Microstructure of the compressed nerve roots were revealed under a transmission electron microscope. BDNF protein expression in the compressed nerve roots and mRNA expression in right solues were detected with enzyme-linked immunosorbent assay (ELISA) and real-time quantitative reverse transcription-polymerase chain reaction respectively. The protein expression level of BDNF in compressed nerve root in the untreated group was slightly changed with no significance as compared with sham-operated group. BDNF protein expression in the Yiqi Huayu Decoction group was gradually increased and reached the peak level after 60-day treatment with significant difference as compared with the untreated group (P<.01). The expression of BDNF mRNA in soleus was significantly up-regulated 10 days after nerve root injury (P<0.01), and was slowly decreased to the normal level at day 60 (P>0.05). Compared with the untreated group, BDNF mRNA expression in soleus muscle in the Yiqi Huayu Decoction group was significantly down-regulated after 10-day treatment (P<0.01), while was increased after 60-day treatment (P<0.01). Yiqi Huayu Decoction can inhibit the increase of BDNF expression at the early phase after nerve injury, which may have something to do with alleviation of neuropathic pain, and increase BDNF expression at the late phase to promote nerve regeneration.