p38MAPK mRNA Research Articles

Effect of Plastrum Testudinis Extracts on the Proliferation and Osteogenic Differentiation of rBMSCs by Regulating p38 MAPK-Related Genes.

Extracts from plastrum testudinis (PTE) are active compounds that have been used to treat bone diseases in traditional Chinese medicine for thousands of years. In previous studies, we demonstrated their effects on glucocorticoid-induced osteoporosis both in vivo and in vitro. However, the mechanisms by which PTE regulates the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) in vitro remain poorly understood. In this study, rBMSCs were treated with medium (CON), PTE, osteogenic induction (OI), and a combination of PTE and OI (PTE+OI) over a 21-day period. We found that PTE significantly promoted rBMSCs osteogenic differentiation and mineralisation after 21 days of culturing. Moreover, PTE+OI further enhanced the differentiation and mineralisation process. PTE upregulated STE20, IGF1R, and p38 MAPK mRNA expression and downregulated TRAF6 mRNA expression. The extracts inhibited TRAF6 protein expression and promoted STE20, IGF1R, and phosphorylated p38 MAPK protein expression. Our results imply that PTE promotes the proliferation and osteogenic differentiation of rBMSCs by upregulating p38 MAPK, STE20, and IGF1R and downregulating TRAF6 expression, which may provide experimental evidence of the potential of PTE in the treatment of osteoporosis.

The influence of aerobic exercise on the structure, function and expression of energy metabolism-related molecules in skeletal muscle of aged rats

Objective To examine the effects of aerobic exercise on muscle morphology, oxidation resistance, energy metabolism, mitochondrial biogenesis-related signal transduction pathway molecules, and energy metabolism-related enzyme proteins expression in aged rats and to explore the molecular mechanisms of skeletal muscle aging and aerobic exercise intervention. Methods Male Sprague Dawley rats were divided into a normal group, a model group, and an exercise group, with the latter two groups receiving a subcutaneous injection of 10% D-galactose(0.1 mg·g-1·d-1)for 10 weeks.Then aerobic and anaerobic exercise capacity of each group were tested by treadmill; the cross-sectional area of rats' gastrocnemius was examined via light microscopy; levels of serum superoxide dismutase(SOD)activity and malondialdehyde(MDA)were determined by spectrophotometry; serum β-Galase levels were tested by enzyme-linked immunosorbent assay(ELISA); levels of adenosine triphosphate(ATP), adenosine diphosphate(ADP)and adenosine monophosphate(AMP)in gastrocnemius tissue were measured by high performance liquid chromatography(HPLC)-ultraviolet detection; expression levels of mitochondrial biosynthesis-related signal transduction molecules(such as p38MAPK, CaMKⅡ, PGC-1α, MEF2C, COXⅣ and ATPase)were tested by real-time RT-PCR. Results Compared with the normal group, the model group's cross-sectional area of muscle fibers, serum SOD activity, ATP content, and EC values all markedly decreased, while serum MDA, β-Galase, AMP content in skeletal muscle significantly increased(all P<0.05). Levels of mRNA expression of CaMKⅡ, MEF2C, ATPaseβ, COXIV, and PCG-1 alpha in model rats notably decreased(all P<0.05), whereas mRNA expression levels of P38MAPK significantly increased(P<0.05). Compared with the model group, aerobic exercise increased the cross-sectional area of muscle fibers(P<0.01)and serum SOD activity(P<0.05), reduced serum MDA content(P<0.01), clearly increased CaMKⅡ and ATPaseβ mRNA expression(both P<0.01), and reduced the expression of P38MAPK mRNA(P<0.01). Conclusions Aerobic exercise can improve the activity of SOD, reduce the accumulation of oxygen free radicals, resist oxidation damage, correct the abnormal activity of P38MAPK, and promote the expression of mitochondrial biosynthesis-related signal molecules, such as CaMKⅡ, PCG-1 alpha, and MEF2C, thus boosting the expression and activity of energy metabolism enzyme protein COX Ⅳ, ATPase, promoting energy metabolism, improving the production of ATP, and slowing down the ageing process of the body and skeletal muscle. Key words: Aerobic exercise; Aging; Skeletal muscle; Antioxidant capacity; Energy metabolism; Mitochondrial biogenesis

PO-165 Effects of Living-High Training-Low on HIF-1α Transcriptional Regulatory Factors MAPKs mRNA in Gastrocnemius of Rats

Objective To evaluate the effects of Living-High Training-Low on HIF-1α transcriptional regulatory factors MAPKs mRNA in gastrocnemius of Rats.&#x0D; Methods After adaptive training, 40 8-weeks-old male SD rats were divided into living-low quiet control group (LC), living-low training-low group (LoLo), living-high quiet control group (HC), living-high training-low group (HiLo). All living-high groups stayed in the environment with 13.6% oxygen concentration, about altitude of 3500 m, for 12h/day. All training groups underwent treadmill training with 35m/min for 1hour/day, 5days/week. 4 weeks later, the gastrocnemius was sampled 24 hours after the last training. The ERK, p38MAPK, JNK and HIF-1α mRNA genes expressions in gastrocnemius were measured by real-time quantitative PCR.&#x0D; Results The gastrocnemius ERK mRNA of HiLo group was significantly higher than LC (P&lt;0.01), LoLo and HC groups (P&lt;0.05). The p38MAPK mRNA of HiLo group was significantly higher than LC and LoLo groups (P&lt;0.01 and P&lt;0.05), and there was no significant difference between HiLo and HC group (P&gt;0.05). The JNK and HIF-1α mRNA of HiLo group were significantly higher than other groups (P&lt;0.01).&#x0D; Conclusions Living-High Training-Low significantly raise ERK、p38MAPK、JNK and HIF-1α gene expression in gastrocnemius of Rats. ERK, p38MAPK and JNK may be one of the transcription factors regulating HIF-1α mRNA expression in Living-High Training-Low in gastrocnemius of Rats.

Chromium malate alleviates high-glucose and insulin resistance in L6 skeletal muscle cells by regulating glucose uptake and insulin sensitivity signaling pathways.

Previous study revealed that chromium malate improved the regulation of fasting blood glucose and insulin resistance in type 2 diabetic rats. In this study, the effect of chromium malate on anti-high-glucose and improve insulin resistance activities in L6 skeletal muscle cells with insulin resistance and its acting mechanism were investigated. Chromium malate showed direct anti-high-glucose activity in vitro. The glucose levels had a significant downward trend compared to chromium trichloride. Compared with model group, chromium malate could significantly promote the secretion levels of GLUT-4, Akt, Irs-1, PPARγ, PI3K and p38-MAPK, promote AMPKβ1 phosphorylation, and reduced the level of p-Irs-1 in L6 cells with insulin resistance. And the relate mRNA expression of chromium malate was significantly increased. Chromium malate is more effective at improving the related proteins and mRNA expression than those of chromium trichloride and chromium picolinate. Pretreatment with the specific p38MAPK inhibitor completely inhibited the GLUT-4 and Irs-1 proteins and mRNA expression induced by the chromium malate when compared with model group, but GLUT-4 and Irs-1 proteins and mRNA expression was partially inhibited after inhibiting p38MAPK/PI3K expression. The results suggested that chromium malate had a beneficial influence on the improvement of controlling glucose levels and insulin resistance in L6 cells with insulin resistance by regulating proteins production and genes expression in glucose uptake and insulin sensitivity signaling pathways. The signaling pathways of glucose uptake and insulin sensitivity. This study shown that chromium malate could significant increase in the production levels of GLUT-4, p-AMPKβ1, Akt, Irs-1, PPARγ, PI3K and p38-MAPK proteins and mRNA in L6 cells with insulin resistant. Pretreatment with the specific p38MAPK inhibitor completely inhibited the GLUT-4 and Irs-1 proteins and mRNA expression induced by the chromium malate compared to model group, but the proteins and mRNA were partially inhibited after inhibiting p38MAPK/PI3K. Therefore, chromium malate had beneficial influence on improvement of controlling glucose levels and insulin resistant in L6 cells by regulating proteins production and genes expression in glucose uptake and insulin sensitivity signaling pathways. The key proteins of glucose uptake and insulin sensitivity signaling pathways were p38MAPK, PI3K and PPARγ.

Effects of inhibiting peritoneal macrophage caspase recruitment domain-containing protein 9 on inflammatory reaction of severe acute pancreatitis in rats

Objective To investigate the effects of caspase recruitment domain-containing protein 9 (CARD9) expression in peritoneal macrophages on severe acute pancreatitis (SAP) in rats and its mechanism. Methods A total of 60 male Sprague Dawley rats were divided into control group (n=6), SAP group (n=18), small interfering RNA (siRNA) control group (n=18) and siRNA CARD9 group (n=18). SAP rat models were established. At three, six and twelve hours after the models were established, ascites was collected, peritoneum was lavaged and peritoneal macrophages were isolated and cultured. The expressions of CARD9, nuclear factor-kappaB (NF-κB), p38 mitogen-activated protein kinase (p38MAPK) at mRNA level in peritoneal macrophages was measured by real-time polymerase chain reaction (RT-PCR). The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). LSD or Tamhane′s T2 methods were performed for statistical analysis. Results At three, six and twelve hours after the models were established, CARD9 mRNA levels of peritoneal macrophages in SAP group were 1.63±0.05, 1.68±0.24 and 2.61±0.02, respectively, which were all higher than that of control group (1.01±0.23), and the differences were statistically significant (t=25.97, 6.86 and 131.59; all P<0.05); the levels of CARD9 mRNA of siRNA CARD9 group were 1.45±0.02, 1.24±0.03 and 1.63±0.03, respectively, which were lower than that of SAP group at the same time points, and the differences were statistically significant (t=-7.81, -4.46 and -62.13; all P<0.05). At three, six and twelve hours after the models were established, the mRNA levels of NF-κB and p38MAPK of peritoneal macrophages of rats in SAP group were 1.51±0.08, 1.81±0.10, 2.30±0.05 and 1.37±0.13, 1.69±0.18, 2.42±0.23, respectively, which were higher than those of control group (1.00±0.01, 1.03±0.08), and the differences were statistically significant (tNF-κB=15.10, 19.95 and 60.36; tp38MAPK=5.37, 8.34 and 14.11; all P<0.05); the levels of NF-κB mRNA in siRNA CARD9 group were 1.38±0.05, 1.57±0.06 and 1.76±0.09, respectively, which were lower than that of SAP group at the same time points, and the differences were statistically significant (t=-3.32, -5.07 and -12.70; all P<0.05). At six and twelve hours after the models were established, the p38MAPK mRNA levels of siRNA CARD9 group were 1.50±0.10 and 2.00±0.09, respectively, which were lower than that of SAP group, and the differences were statistically significantly (t=-2.30 and -4.17, both P<0.05). At three, six and twelve hours after the models were established, the levels of TNF-α, IL-1β and IL-6 in peripheral blood of SAP group were (53.49±21.64) pg/mL, (108.62±22.76) pg/mL and (139.00±15.35) pg/mL; (43.86±18.30) pg/mL, (87.51±17.10) pg/mL and (117.27±14.57) pg/mL; (78.38±32.70) pg/mL, (156.39±30.56) pg/mL and (209.56±26.09) pg/mL, respectively, which were higher than those of control group ((2.79±1.17), (7.13±4.52), (12.73±8.08) pg/mL), and the differences were statistically significant (tTNF-α=5.73, 11.37 and 21.69; tIL-1β=4.77, 11.13 and 17.68; tIL-6=4.77, 11.32 and 17.68; all P<0.05). At six and twelve hours after the models were established, the levels of TNF-α, IL-1β and IL-6 of siRNA CARD9 group were (75.73±16.93) pg/mL, (108.23±14.02) pg/mL; (63.05±11.98) pg/mL, (91.56±14.28) pg/mL and (112.67±21.40) pg/mL, (163.62±25.51) pg/mL, respectively, which were lower than those of SAP group, and the differences were statistically significant (tTNF-α=-2.84, -3.63; tIL-1β=-2.88, -3.09; tIL-6=-2.88, -3.09; all P<0.05). Conclusions There are CARD9-related NF-κB and p38MAPK pathways in peritoneal macrophages of SAP rats. Intervention of the expression of CARD9 in peritoneal macrophages, especially at early stage of SAP may relieve the inflammation reaction and pancreatic injury, which may provide a new method for SAP treatment. Key words: Macrophages, abdominal cavity; NF-kappaB; p38 mitogen-activated protein kinases; RNA, small interfering; Rats; Pancreatitis, severe acute; Caspase recruitment domain-containing protein 9

Exogenous Hydrogen Sulfide Protects SH-SY5Y Cells from OGD/RInduced Injury.

Cerebral ischemia reperfusion (CIR) injury is the main pathological mechanism of ischemia stroke, and oxidative stress (OS) plays a vital role during the process of CIR. Recent studies have found that OS can activate MAPK pathway, which mainly functions as a mediator of cellular stresses. It revealed that H2S may exhibit multiple biological functions as a neuroprotector through protecting against OS-induced neuronal injury. But how H2S can attenuate ischemia/reoxygenation injury through MAPK signaling is not clear. The protective mechanism of exogenous hydrogen sulfide (H2S) on oxygen glucose deprivation/reoxygenation (OGD/R) at physiological doses in SH-SY5Y cells were investigated in this study. DCFH- DA probe is used to measure intracellular ROS, Real-time PCR and Western blot are used to detect the levels of p38 MAPK, ERK3 and Nrf2. CCK-8 is used to detect cell viability. Comparing with the normal control group, the expressions of ERK3, p38MAPK and Nrf2 mRNA in other three groups were downregulated at the beginning of 24h. After 24h, the expressions of ERK3, p38MAPK and Nrf2 were upregulated significantly compared with the normal control group. When the cells were pretreated with 200μM NaHS at the beginning of 24h, the expressions of ERK3, p38MAPK and Nrf2 were decreased much more significantly than OGD/R and NaHS groups. The present study demonstrated that exogenous H2S exerts a protective effect against OGD/R-induced injury by enhancing the activation of the ERK3, p38MAPK and Nrf2 mRNA.

Structural characterization of a pectic polysaccharide from Codonopsis pilosula and its immunomodulatory activities in vivo and in vitro

A pectic polysaccharide (named as CPP1c) extracted from Codonopsis pilosula was evaluated for its structural features and potential of immune-modulating activities in an aging mouse model of senescence accelerated mouse prone 8 (SAMP8) in vitro and in vivo. The relative molecular weight and the absolute molecular weight of CPP1c were 1.26×105Da and 1.49×105Da, respectively. Investigation of structural features by a combination of chemical and instrumental analysis showed CPP1c was composed of →1)-α-l-Rhap-(2,4→, →1)-α-l-Araf-(5→, →1)-α-d-Galp-(6→ and →1)-α-d-GalpA-(4→ in a molar ratio of 3:1:2:33. CPP1c could promote lymphocyte proliferation, modulate the percentage of CD4+, CD8+, CD28+ and CD152+ T cells and enhance the production of IL-2, TNF-α and IFN-γ. Moreover, PCR assay revealed CPP1c augmented the expressions of CD28, PI3K and p38MAPK mRNA, and the increase of protein expressions of the same genes was also confirmed by western blot analyses. In addition, CPP1c had the potential of promoting the homing of lymphocytes. Taking all factors into consideration, we deduced CPP1c might exert its immunostimulating potency via promoting T cell activation by TCR/CD28 signaling pathways.

Bufei Yishen Granules Combined with Acupoint Sticking Therapy Suppress Inflammation in Chronic Obstructive Pulmonary Disease Rats: Via JNK/p38 Signaling Pathway.

The present study was initiated to explore the mechanism of the effects of Bufei Yishen granules combined with acupoint sticking therapy (Shu-Fei Tie) on inflammation regulated by c-Jun N-terminal kinase (JNK) and p38 MAPK signaling in COPD rats. Seventy-two rats were divided into healthy control (Control), Model, Bufei Yishen (BY), acupoint sticking (AS), Bufei Yishen + acupoint sticking (BY + AS), and aminophylline (APL) groups (n = 12 each). COPD rats were exposed to cigarette smoke and bacteria and were given the various treatments from weeks 9 through 20; all animals were sacrificed at the end of week 20. MCP-1, IL-2, IL-6, and IL-10 concentrations in BALF and lung tissue as well as JNK and p38 mRNA and protein levels in lung were measured. The results showed that all the four treatment protocols (BY, AS, BY + AS, and APL) markedly reduced the concentrations of IL-2, IL-6, and MCP-1 and levels of JNK and p38 MAPK mRNA, and the effects of Bufei Yishen granules combined with acupoint sticking therapy were better than acupoint sticking therapy only and aminophylline. In conclusion, the favorable effect of Bufei Yishen granules combined with Shu-Fei Tie may be due to decreased inflammation through regulation of the JNK/p38 signaling pathways.

Notoginsenoside R&amp;lt;sub&amp;gt;1&amp;lt;/sub&amp;gt; Attenuates Hypoxia and Hypercapnia-Induced Vasoconstriction &amp;lt;i&amp;gt;In Vitro&amp;lt;/i&amp;gt; by Reducing the Expression of p38

Notoginsenoside R1, the main active ingredient of Panax notoginseng saponins (PNS), has been proposed to play fatal roles in the development of hypoxic hypercapnia-induced pulmonary vasoconstriction (HHPV). Subsequently, pulmonary arterial smooth muscle cells (PASMCs) lead to pulmonary vascular system remodeling and chronic pulmonary disease in the development of HHPV. Despite considerable studies have contributed to pulmonary disease, the mechanism of how Notoginsenoside R1 affects HHPV remains unclear. In this view, we will discuss the effect of notoginsenoside R1 by investigating the expression of p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in PASMCs under hypoxia and hypercapnia condition. The third order PASMCs of Sprague Dawley (SD) rats were cultured with various concentrations (8, 40, 100 mg/L, respectively) of Notoginsenoside R1. Our data showed that the protein and mRNA expression levels of p-38 MAPK were higher in hypoxic hypercapnia group compared with hypoxic DMSO and normoxia control groups (p 1 treatment groups, the level of p-p38 MAPK protein and p38 MAPK mRNA were significantly decreased with different degrees (p 1 treatment may contribute to attenuate HHPV via decreasing the protein and mRNA expression levels of p-38 MAPK.

Low-frequency stimulation induces a durable long-term depression in young adult hyperthyroid rats: the role of p38 mitogen-activated protein kinase and protein phosphatase 1.

Long-term potentiation and long-term depression (LTD) are cellular mechanisms of learning and memory in the mammalian brain. We have previously shown that adult hyperthyroid rats showed a delay in the acquisition of a place learning task and attenuated long-term potentiation. However, changes in LTD in hyperthyroidism remain unclear. Rats were administered 0.2 mg/kg/day of L-thyroxine for 21 days starting at postnatal day 40 to induce hyperthyroidism. LTD was induced in the dentate gyrus using low-frequency stimulation (LFS) of the perforant pathway. The mRNA expressions of p38 mitogen-activated protein kinase (p38-MAPK) and protein phosphatase 1 (PP1) were evaluated using a quantitative reverse transcriptase PCR. In control rats, a standard LFS protocol induced a slight depression of the population spike (PS) amplitude during the induction phase of LTD (76±13% vs. baseline), but a small potentiation of the PS amplitude was observed in the early (107±18%) and late (111±20%) phases of LTD. Interestingly, in the hyperthyroid rats, the same LFS protocol induced a reliable LTD in the dentate gyrus of the hippocampus as evidenced by a marked depression in the PS amplitude during the induction (54±6% vs. baseline) and the early phases (56±8%) of LTD. Elevated mRNA levels of p38-MAPK and PP1 were observed in the hippocampus of the LFS-treated hyperthyroid rats compared with the hippocampus of the vehicle-treated hyperthyroid rats. No significant change in p38-MAPK or PP1 mRNA expression was observed in the euthyroid rats. The present study shows that a standard LFS protocol can induce a durable depression of synaptic strength and an upregulation of PP1 and p38-MAPK mRNA in hyperthyroid rats. We conclude that hyperthyroidism can induce molecular changes associated with degeneration of the hippocampus. The relationship between the levels of thyroid hormone and dementia requires further investigation.

Radix Astragali Stimulates p38 MARK Phosphorylation in Pediatric Patients with β-Thalassemia.

A previous study conducted by our group demonstrated that Radix Astragali compounded with Codonopsis pilosula and Plastrum testudinis was effective in treating pediatric β-thalassemia in a randomized, controlled clinical trial. However, the mechanism of action that underpins this treatment remains to be elucidated. Blood was collected from patients participating in this clinical trial and nucleated red blood cell-enriched mononuclear cells were isolated to facilitate the extraction of RNA and protein. RT-PCR was used to monitor the expression of globin genes and p38 MAPK, and total and phosphorylated p38 MAPK expression was assessed using Western blot analysis. Expression of α-, β-, and Aγ-globin mRNAs was not significantly affected following treatment with R. Astragali or the compounded formulation. However, Gγ-globin mRNA levels increased significantly in both treatment groups (when compared with pretreatment levels) following 12 weeks of treatment. Moreover, posttreatment Gγ-globin expression was significantly higher in both treatment groups compared with the control group. Although neither p38 MAPK mRNA nor protein levels were affected by the treatments, posttreatment phosphorylation of p38 MAPK was significantly increased in the R. Astragali and compounded formulation groups compared with the control group. These data suggest that the molecular mechanisms that underpin the efficacious use of R. Astragali (and its compounded formulation) in pediatric β-thalassemia treatment facilitate the induction of Gγ-globin expression following activation of p38 MAPK.

Signal pathway defects of Toll-like receptors 2-p38 mitogen activated protein kinase signal pathway defects and immune suppression in children with measles

Objective To investigate the expression of Toll-like receptors 2(TLR2)mRNA, p38 mitogen activated protein kinase(p38 MAPK)mRNA and cytokine in peripheral blood of children with measles and to study the effect and possible mechanism for TLR2-p38 MAPK signal pathway defects on immune suppression in the children with measles during acute phase. Methods Thirty children with measles hospitalized in the department of infectious diseases from June 2012 to July 2013 were enrolled into the measles group, and 30 healthy children were chosen as the healthy control group.The mRNA expressions of TLR and p38 MAPK in peripheral blood mononuclear cells (PBMC)were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein levels of interferon-γ(IFN-γ), tumor necrosis factor-β(TNF-β), interleukin(IL)-12, IL-6 and IL-10 in plasma were measured by using enzyme linked immunosorbent assay (ELISA), and flow cytometry (FCM)was applied to detect the percentage of lymphocyte subpopulation.The serum IgG, IgA and IgM levels were detected by velocity scatter turbidimetry. Results (1)The expressions of TLR2 mRNA and p38 MAPK mRNA in the measles group were both significantly lower than those in the healthy control group (all P 0.05). Conclusions The mRNA expressions of TLR2 and p38 MAPK are low in PBMC in the measles children during acute phase.There are different degrees of suppression of cell immunity, humoral immune and cytokines disorder in children with measles.Defects of TLR2-p38 MAPK signal pathway may cause the formation of measles immune suppression. Key words: Measles; Toll-like receptors 2; p38 mitogen activated protein kinase; Immune suppression; Child

Effect of different concentrations of oxygen on inflammatory response in lipopolysaccharide-induced alveolar typeIIcells

Objective To investigate the secretion of inflammatory cytokines in lipopolysaccharide(LPS)-induced aveolar epithelium typeⅡcells(AT-Ⅱ) cells exposed to various oxygen concentrations, and the possible mechanism. Methods Primary cultured AT-Ⅱcells were randomly divided into five groups: A, B, C, D, and E, and then placed in sterilized incubators: 37℃, 21% O2, 5% CO 2; 37℃, 21% O2, 5% CO 2; 37℃, 40% O2, 55% N2, 5% CO 2; 37℃, 60% O 2, 35% N2, 5% CO2; 37℃, 90% O 2, 5% N 2, 5% CO 2.Towenty-four h later, the supernatant fluid was used to assay the content of interleukin-8(IL-8) by enzyme linked immunosorbent assay(ELISA).The expression of p38 mitogen-activated protein kinase(p38 MAPK) m RNA and protein was detected in AT-Ⅱcells by reverse transcription-polymerase chain reaction and Western blotting. Results ELISA, reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blotting showed that the content of IL-8 was(40. 22±7. 25),(109. 35±10. 19),(213. 98 ±30. 51),(369. 71±21. 73)and(489. 32±42. 35) ng/L, p38MAPK mRNA expression level was 0. 066±0. 010, 0. 163±0. 008, 0. 228±0. 027, 0. 268±0. 016 and 0. 292±0. 010, and p38MAPK protein expression level was 0. 055±0. 005, 0. 124±0. 006, 0. 182±0. 009, 0. 227±0. 013 and 0. 303±0. 010 in groups A, B, C, D and E respectively.As compared with group A, the content of IL-8 in the supernatant fluid,and the expression of p38MAPK m RNA and protein were significantly increased in groups B, C, D and E(P< 0. 05).As compared with groups B, the content of IL-8 in the supernatant fluid, and the p38 MAPK m RNA and protein expression level in groups C, D and E were gradually increased with the difference being statistically significant(P< 0. 05).In different oxygen concentration groups, with the increases in oxygen concentrations, the content of IL-8 in the supernatant fluid, and the expression of p38MAPK m RNA and protein expression level were gradually increased with the difference being statistically significant(P< 0. 05). Conclusion 40%-90% oxygen could concentrasion-dependently increase the LPS-induced secretion of IL-8 in AT-Ⅱcells probably by up-regulating the intracellular p38 MAPK expression. Key words: Hyperoxia-induced lung injury; Aveolar epithelium typeⅡcells; Lipopolysaccharide; Interleukin-8; p38 mitogen-activated protein kinase

GLP-1 receptor activation effects the p38MAPK signal pathway in hepatic stellate cells

To investigate the effects of activation of the GLP-1 receptor on the p38MAPK signaling pathway in hepatic stellate cells (HSCs). HSCs were isolated and identified according to morphological features; the levels of GLP-1R protein were determined by western blotting.The HSCs were randomly divided into a control grouP (normal saline treatment) and experimental grouP(liraglutide treatment); after 120 hours, the expression of p38MAPK mRNA was examined by RT-PCR and of phosphorylated (p)-p38MAPK protein was detected by western blotting. GLP-1R proteins were detected in the HSCs. Compared with the control group, the experimental group showed significantly decreased p38MAPK mRNA and p-p38MAPK protein (both P < 0.01). The p38MAPK signaling pathway could be down-regulated when GLP-1R is activated in HSCs.

Effect of Repeated Electroacupuncture Intervention on Hippocampal ERK and p38MAPK Signaling in Neuropathic Pain Rats.

Results of our past studies showed that hippocampal muscarinic acetylcholine receptor (mAChR)-1 mRNA and differentially expressed proteins participating in MAPK signaling were involved in electroacupuncture (EA) induced cumulative analgesia in neuropathic pain rats, but the underlying intracellular mechanism remains unknown. The present study was designed to observe the effect of EA stimulation (EAS) on hippocampal extracellular signal-regulated kinases (ERK) and p38 MAPK signaling in rats with chronic constrictive injury (CCI) of the sciatic nerve, so as to reveal its related intracellular targets in pain relief. After CCI, the thermal pain thresholds of the affected hind were significantly decreased compared with the control group (P < 0.05). Following one and two weeks' EAS of ST 36-GB34, the pain thresholds were significantly upregulated (P < 0.05), and the effect of EA2W was remarkably superior to that of EA2D and EA1W (P < 0.05). Correspondingly, CCI-induced decreased expression levels of Ras, c-Raf, ERK1 and p-ERK1/2 proteins, and p38 MAPK mRNA and p-p38MAPK protein in the hippocampus tissues were reversed by EA2W (P < 0.05). The above mentioned results indicated that EA2W induced cumulative analgesic effect may be closely associated with its function in removing neuropathic pain induced suppression of intracellular ERK and p38MAPK signaling in the hippocampus.

Effect of curcumin on p38MAPK expression in DSS-induced murine ulcerative colitis.

The aim of this study was to determine the therapeutic effect of curcumin on dextran sulfate sodium-induced ulcerative colitis (UC) and to explore the related mechanism. Sixty mice were randomly divided into 6 groups. A group was the normal control group; B group was the model group; C group was the 1.5 mg/kg dexamethasone group based on the B group; and D, E and F groups were 15, 30, and 60 mg/kg curcumin groups, respectively, based on the B group. The mice were killed 7 days after treatment; the expression of TNF-α and MPO in colon tissue was determined with ELISA, and colon p-p38MAPK and p38MAPK mRNA expression was evaluated by immunohistochemistry and RT-PCR, respectively. In the C, D, E, and F groups, TNF-α and MPO levels significantly decreased (P < 0.05), and the expression of p-p38MAPK also significantly decreased (P < 0.01). The expression of p38MAPK mRNA in the C, D, E, and F groups decreased (P < 0.01), and there was a statistically significant difference between the E and F groups (P < 0.01). Curcumin had a therapeutic effect, which probably played a role in UC treatment by inhibiting the p38MAPK signaling pathway, thereby reducing the release of TNF-α.

Effect of panax notoginseng saponins injection on the p38MAPK pathway in lung tissue in a rat model of hypoxic pulmonary hypertension.

To investigate the effect of panax notoginseng saponins (PNS) injection on pulmonary artery pressure and the expression of p38MAPK in lung tissue of rats subjected to chronic hypoxia. Thirty adult male Sprague Dawley rats were randomly divided into three groups (ten in each group): rats in control group were exposed to normoxic condition and the rats in hypoxia group and PNS group were subjected to 4-week hypoxia, and PNS injection (50 mg · kg(-1) · d(-1)) was administrated intraperitoneally at 30 min in the PNS group daily before the rats were kept in the hypoxic chamber, while rats in the other two groups received equal dose of normal saline instead. After chronic hypoxia, mean pulmonary artery pressure (mPAP) and mean carotid artery pressure (mCAP) were measured. The heart and lung tissues were harvested, and right ventricle (RV) and left ventricle plus ventricular septum (LV+S) were weighed to calculate the ratio of RV/(LV+S). The expression of p38MAPK mRNA was determined by reverse transcription-polymerase chain reaction, the quantity of phosphorylated p38MAPK (p-p38MAPK) in rat lung tissues and pulmonary arterioles was determined by Western blot and immunohistochemistry. Compared with the control group, mPAP and the ratio of RV/(LV+S) in the hypoxia group were increased, the expression of p-p38MAPK in pulmonary arterioles and p38MAPK mRNA in the lung were higher (P<0.05). The changes of these parameters in the hypoxia group were significantly attenuated by PNS treatment (P<0.05). PNS injection was shown to prevent hypoxic pulmonary hypertension at least partly by regulating p38MAPK pathway.

Effects of reduced β2-glycoprotein I on the expression of aortic matrix metalloproteinases and tissue inhibitor matrix metalloproteinases in diabetic mice.

BackgroundReduced β2-glycoprotein I (reduced β2GP I), which has free sulfhydryl groups, is present in plasma and serum; it can protect vascular endothelial cells from damage due to oxidative stress in vitro. We investigated the effects of reduced β2GP I on the expression of various matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the aortas of diabetic mice.MethodsWe provided 120 female 8-week-old Balb/c mice with a high sugar, high fat diet. After 8 weeks they were injected with streptozotocin to induce diabetes. We treated mice in the mono dose groups with β2GP I, reduced β2GP I, or phosphate-buffered saline (PBS) on day 1 and fed them for 3 weeks. The mice in the complex dose groups were treated with β2GP I, reduced β2GP I, or PBS on days 1 and 22 and fed for 6 weeks. Control mice were given a standard chow diet. Blood lipids were measured at the end of 3 or 6 weeks, and aortas removed to observe morphological and molecular biological changes.ResultsThe low-density lipoprotein cholesterol levels in mice of the reduced β2GP I group were lower than those in the diabetic group. Aortic lipid deposition in the reduced β2GP I group was significantly less than in the diabetic control group. In the aortas, reduced β2GP I decreased MMP2/TIMP2 mRNA and protein expression levels, and MMP9/TIMP1 expression levels compared with those in diabetic controls. Reduced β2GP I down-regulated p38 mitogen-activated protein kinase (p38MAPK) mRNA expression and phosphorylated p38MAPK protein expression compared with those in diabetic controls of the complex dose group.ConclusionsReduced β2GP I plays a role in diabetic mice related to vascular protection, inhibiting vascular lipid deposition, and plaque formation by reducing MMPs/TIMPs expression through down-regulation of the p38MAPK signaling pathway.

NADPH oxidase hyperactivity induces plantaris atrophy in heart failure rats

BackgroundSkeletal muscle wasting is associated with poor prognosis and increased mortality in heart failure (HF) patients. Glycolytic muscles are more susceptible to catabolic wasting than oxidative ones. This is particularly important in HF since glycolytic muscle wasting is associated with increased levels of reactive oxygen species (ROS). However, the main ROS sources involved in muscle redox imbalance in HF have not been characterized. Therefore, we hypothesized that NADPH oxidases would be hyperactivated in the plantaris muscle of infarcted rats, contributing to oxidative stress and hyperactivation of the ubiquitin-proteasome system (UPS), ultimately leading to atrophy. MethodsRats were submitted to myocardial infarction (MI) or Sham surgery. Four weeks after surgery, MI and Sham groups underwent eight weeks of treatment with apocynin, a NADPH oxidase inhibitor, or placebo. NADPH oxidase activity, oxidative stress markers, NF-κB activity, p38 MAPK phosphorylation, mRNA and sarcolemmal protein levels of NADPH oxidase components, UPS activation and fiber cross-sectional area were assessed in the plantaris muscle. ResultsThe plantaris of MI rats displayed atrophy associated with increased Nox2 mRNA and sarcolemmal protein levels, NADPH oxidase activity, ROS production, lipid hydroperoxides levels, NF-κB activity, p38 MAPK phosphorylation and UPS activation. NADPH oxidase inhibition by apocynin prevented MI-induced skeletal muscle atrophy by reducing ROS production, NF-κB hyperactivation, p38 MAPK phosphorylation and proteasomal hyperactivity. ConclusionOur data provide evidence for NADPH oxidase hyperactivation as an important source of ROS production leading to plantaris atrophy in heart failure rats, suggesting that this enzyme complex plays key role in skeletal muscle wasting in HF.

Effect of Antrodia cinnamomea on gene expression related to aortal endothelial injury in rats with hyperlipidemia

To investigate the effect of Antrodia cinnamomea on gene expression related to aortal endothelial injury of rats with hyperlipidemia. Fifty SD rats were randomly divided into five groups: the normal control group (NG), the model group (MG), the antrodia cinnamomea groups of low, middle and high doses (AC-LG, AC-MG, AC-HG, 250, 500, 1 000 mg x kg(-1)). The rats were fed with high-fat diets to establish the hyperlipidemia model. After the drug administration for 10 weeks, their serum lipid, SOD, MDA and ox-LDL, LOX-1, P38 MAPK and NF-kappaB mRNA and protein expression were respectively determined, and the aortal endothelial injury was observed under electron microscope. In the model group, the contents of TC, TG and LDL-C significant increased (P < 0.01), whereas the content of HDL-C significant decreased (P < 0.01). Compared with the model group, both the AC-M group and the AC-H group showed reduction in endothelial injury and significant decrease in the content of TC, TG and LDL-C (P < 0.05 or P < 0.01). The content of HDL-C increased, but with no significant difference. SOD activity in serum remarkably increased (P < 0.05 or P < 0.01), MDA and ox-LDL levels dramatically decreased (P < 0.05 or P < 0.01). A. cinnamomea can alleviate endothelial lipid injury by inhibiting the expressions of LOX-1, P38MAPK and NF-kappaB in aorta and better protect aortal endothelial cells from oxidative lipid injury.