Role of p 38 MAPK signaling pathway on post-stroke cognitive impairment and pharmacological research progress in the prevention and treatment with traditional Chinese medicine / 中国药房

Post-stroke cognitive impairment （PSCI） refers to a series of syndromes from mild cognitive impairment to dementia caused by stroke. The mitogen-activated protein kinases （MAPK）signaling pathway is a key pathway for transmitting cellular signals in mammals ，and p 38 is a classic branch of it. p 38 MAPK signaling pathway is involved in various pathophysiological processes such as cell growth ，differentiation，apoptosis and inflammatory response in central nervous system diseases. At present ，great progress has been made in clinical and basic experimental studies on prevention and treatment of PSCI by traditional Chinese medicine （TCM），but there is a lack of relevant systematic summary. Therefore ，this article summarizes the role of p 38 MAPK signaling pathway in PSCI and the pharmacological research progress of TCM in prevention and treatment of PSCI through p 38 MAPK signaling pathway.

Effect of Fushengong Prescription on p38 MAPK Signal Pathway of Rats with Chronic Renal Failure / 中国实验方剂学杂志

Objective:To observe the effects of Fushengong prescription on p38 mitogen-activated protein kinase（p38 MAPK） signal pathway of rats with chronic renal failure（CRF），and to explore its mechanism of reducing inflammatory reaction of renal tissues and delaying the progress of renal interstitial fibrosis. Method:The 55 male Sprague-Dawley rats were randomly divided into normal group，model group， and low，medium and high dose groups of Fushengong prescription，with 11 rats in each group.The normal group was routinely reared， and the other four groups of rats were fed a diet containing 0.5% adenine to produce a model of CRF， which was continuously molded for 21 days.After successful modeling，all rats switched to conventional feed.Normal group and model group were given normal saline 20 mL·kg-1，and each group of Fushengong prescription was given 4，8，16 g·kg-1 of water prescription once a day for 30 days.After the experiment，Masson staining was used to observe the degree of renal interstitial fibrosis.The expression of monocyte chemotactic protein-1（MCP-1） in renal tissues was detected by immunohistochemistry. The expression of phosphorylated p38 mitogen-activated protein kinase（p-p38 MAPK） and transformed growth factor-β1（TGF-β1） in renal tissues were detected by Western blot. Result:Compared with normal group，the renal interstitial collagen deposition increased significantly，the average optical density value of MCP-1 and the expression levels of p-p38 MAPK and TGF-β1 also increased significantly in model group （P<0.05）. Compared with model group，the renal interstitial collagen deposition reduced significantly，the average optical density value of MCP-1 and the protein expression levels of p-p38 MAPK and TGF-β1 also decreased significantly in each dose group of Fushengong prescription（P<0.05）. Conclusion:Fushengong prescription can effectively inhibit the expression of related inflammatory factors in the renal tissue of CRF rats，so as to reduce the inflammatory response in the renal tissue and delay the progress of renal interstitial fibrosis，the mechanism of which may be related to inhibit the activation of p38 MAPK signal transduction pathway.

Effect of ferulic acid on pulmonary function in mice with chronic obstructive pulmonary disease and its mechanism / 中国中西医结合急救杂志

Objective To investigate the protective effect of ferulic acid on lung function in mice with chronic obstructive pulmonary disease (COPD) and its possible mechanism. Methods Sixty mice were randomly divided into normal control group, COPD model group, Rofloast group and ferulic acid high, medium and low dose groups, each group with 10 rats, and during administration one rat died in the mode group and was eliminated. The COPD model was duplicated by smoking method; the mice in normal control group were fed normally without any treatment. After modeling for 30 days, normal saline begun to be given to the COPD model group and normal control group; the mice in Rofluast group were given Rofluast 65 μg/kg; ferulic acid 160, 80, 40 mg/kg were given to high, middle and low dose groups respectively. The indexes were determined after consecutive 90 days of treatment, the changes of peak inspiratory flow (PIF) rate, peak expiratory flow (PEF) rate and ventilation volume per minute (MV), mean lining interval (MLI), alveolar destruction index (DI), interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) levels in serum and bronchoalveolar lavage fluid (BALF), the protein expressions and phosphorylation levels of p38 mitogen-activated protein kinases (p38MAPK), extracellular signal-regulated kinase (ERK) and c-Jun amino terminal kinase (JNK) in the pulmonary tissue MAPK signaling pathway were observed in each mouse of various mice groups. Results In the COPD model group, the PIF, PEF, and MV were all significantly lower than those in the normal control group [PIF (mL/s): 2.32±0.18 vs. 3.41±0.12, PEF (mL/s): 2.31±0.22 vs. 2.90±0.15, MV (mL/s): 26.20±2.70 vs. 35.18±2.30); Luofusite and all doses of ferulic acid can increase PIF, PEF, and MV, and the degree of increase in the high dose ferulic acid group was more significant than those in the moderate and low dose ferulic acid groups [PIF (mL/s): 3.24±0.13 vs. 2.88±0.15, 2.51±0.10, PEF (mL/s): 2.81±0.16 vs. 2.66±0.11, 2.58±0.17, MV (mL/s):31.18±1.20 vs. 28.25±2.20, 27.09±1.10]; however, there was no statistical significant difference between the Rofluas group and the ferulic acid groups (all P > 0.05). The levels of the MLI, DI, and inflammatory factors in serum and BALF, and the protein expressions and phosphorylation levels of p38MAPK, ERK, JNK in lung tissue in model group were all significantly higher than those in normal control group [MLI (μm): 52.10±0.26 vs. 21.90±0.14, DI: (60.78±3.32)% vs. (22.47±1.05)%, IL-6 in serum (ng/L): 22.34±4.51 vs. 3.50±1.55, TNF-αin serum (ng/L): 27.11±3.99 vs. 4.66±1.76, IL-6 (ng/L) in BALF: 142.92±20.10 vs. 18.77±4.17, TNF-α(ng/L): 150.16±20.77 vs. 22.01±4.15, P-ERK/ERK (gray value): 0.59±0.03 vs. 0.38±0.05, P-p38MAPK/p38MAPK (gray value): 0.52±0.02 vs. 0.31±0.05, P-JNK/JNK (gray value): 0.56±0.03 vs. 0.25±0.01, all P < 0.05]. The levels of MLI, DI, and inflammatory factors in serum and BALF, p38MAPK, ERK, JNK protein expression and phosphorylation in lung tissue were reduced by Rofluas and various doses of ferulic acid, the reduction levels in the high dose group of ferulic acid were more significant than those in the middle and low dose groups of ferulic acid [MLI (μm): 25.00±0.19 vs. 30.10±0.29, 38.80±0.41, DI: (26.32±3.05)% vs. (29.75±6.17)%, (40.56±5.81)%, IL-6 in serum (ng/L): 9.20±1.87 vs. 12.35±2.16, 18.95±3.12, TNF-α(ng/L): 13.37±2.73 vs. 18.02±2.62, 21.31±3.75, IL-6 (ng/L) in BALF: 64.27±11.72 vs. 99.33±13.02, 120.31±18.02, TNF-α(ng/L): 58.20±10.28 vs. 93.83±16.26, 122.68±14.85, P-ERK/ERK (gray value): 0.43±0.04 vs. 0.46±0.04, 0.52±0.02, P-p38MAPK/p38MAPK (gray value): 0.33±0.03 vs. 0.34±0.03, 0.38±0.02, P-JNK/JNK (gray value): 0.32±0.04 vs. 0.38±0.05, 0.47±0.06). The ferulic acid could improve the inflammatory cell infiltration situation in mice with COPD. Conclusions Ferulic acid can improve pulmonary inflammation in COPD rats. The effective mechanism is possibly related to the inhibition of the protein expressions and phosphorylation levels of the key proteins such as p38MAPK, ERK and JNK in the MAPK signaling pathway.

Effect of Soothing Gan (Liver) and Invigorating Pi (Spleen) Recipes on TLR4-p38 MAPK Pathway in Kupffer Cells of Non-alcoholic Steatohepatitis Rats / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of inflflammatory-mediated toll-like receptor 4 (TLR4)-p38 mitogen-activated protein kinase (p38 MAPK) pathway in Kupffer cells (KCs) of non-alcoholic steatohepatitis (NASH) rats and the intervention effect of soothing Gan (Liver) and invigorating Pi (Spleen) recipes on this pathway.@*METHODS@#After 1 week of acclimatization, 120 Sprague-Dawley male rats were randomly divided into 8 groups using a random number table (n=15 per group): normal group, model group, low-dose Chaihu Shugan Powder (, CHSG) group (3.2 g/kg), high-dose CHSG group (9.6 g/kg), low-dose Shenling Baizhu Powder (, SLBZ) group (10 g/kg), high-dose SLBZ (30 g/kg) group, and low- and highdose integrated recipe (L-IR, H-IR) groups. All rats in the model and treatment groups were fed with a high-fat diet (HFD). The treatments were administrated by gastrogavage once daily and lasted for 26 weeks. The liver tissues were detected with hematoxylin-eosin (HE) and oil red O staining. Levels of liver lipids, serum lipids and transaminases were measured. KCs were isolated from the livers of rats to evaluate the mRNA expressions of TLR4 and p38 MAPK by real-time flfluorescence quantitative polymerase chain reaction, and proteins expressions of TLR4, p-p38 MAPK and p38 MAPK by Western blot. Levels of inflammatory cytokines including tumor necrosis factor α (TNF-α), interleukin (IL)-1 and IL-6 in KCs were measured by enzyme-linked immunosorbent assay.@*RESULTS@#After 26 weeks of HFD feeding, HE and oil red O staining showed that the NASH model rats successfully reproduced typical pathogenesis and histopathological features. Compared with the normal group, the model group exhibited significant increases in body weight, liver weight, liver index, serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol, and aspartate aminotransferase as well as TC and TG levels in liver tissues, and significant decrease in serum level of high-density lipoprotein cholesterol (Plt;0.05 or Plt;0.01), while those indices were significantly ameliorated in the H-IR group (Plt;0.05 or Plt;0.01). Higher levels of TNF-α, IL-1 and IL-6 in KCs were observed in the model group compared with the normal group (Plt;0.01). Significant decreases in TNF-α, IL-1 and IL-6 were observed in the H-SLBZ, H-IR and L-IR groups compared with the model group (Plt;0.05 or Plt;0.01). The mRNA expressions of TLR4 and p38 MAPK and protein expressions of TLR4, p38 MAPK and p-p38 MAPK in KCs in the model group were significantly higher than the normal group (Plt;0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group (Plt;0.05 or Plt;0.01).@*CONCLUSION@#Inflflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38MAPK signaling pathway in KCs for the anti-inflflammatory effect of soothing Gan and invigorating Pi recipes.

A β-catenin/IQGAP1 regulatory feedback loop and its effects on the proliferation of colon cancer cells / 生物医学工程学杂志

The aim of this article is to study the regulatory feedback loop between β-catenin and IQ motif containing GTPase activating protein 1 (IQGAP1), as well as the effect of this regulation loop in colon cancer cell proliferation. Western blot was used to detect the expression of IQGAP1 and β-catenin after changing their expression respectively by transfection in SW1116 cells. CCK-8 cell proliferation assay was used to detect the effect of IQGAP1 involved in the proliferation of SW1116 cells promoted by β-catenin. The results of Western blot indicated that β-catenin could positively regulate IQGAP1, while IQGAP1 silencing could up-regulate β-catenin, forming a negative feedback loop. The results of CCK-8 showed that IQGAP1 silencing inhibited β-catenin-mediated proliferation in SW1116 cells. In conclusion, our research reveals a negative regulatory feedback loop between β-catenin and IQGAP1 which has a remarkable effect on the proliferation ability of colon cancer cells.

Role of MAPK Signaling Pathways in Regulating the Hydrophobin Cryparin in the Chestnut Blight Fungus Cryphonectria parasitica

We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in Cryphonectria parasitica. Mutation of the CpSlt2 gene, an ortholog of yeast SLT2 in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the CpBck1 gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the cpmk1 gene, an ortholog of yeast HOG1, showed decreased cryparin production. However, mutation of the cpmk2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. In silico analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both CpRlm1, an ortholog of yeast RLM1 in the CWI pathway, and cpst12, an ortholog of yeast STE12 in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in C. prasitica. However, the transcription of CpMcm1, an ortholog of yeast MCM1, did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.

The role of microRNA -155,microRNA -222 and mitogen -activated protein kinase signaling pathway in ven-tricular septal defect / 中华实用儿科临床杂志

Objective To explore the expression and clinical significance of microRNA (miR)-155 and miR -222 in plasma of patients with ventricular septal defect(VSD),and to analyze the possible mechanism.Methods A total of 20 children with VSD who received treatment at the Department of Cardiovascular Surgery from August 2012 to June 2013 were enrolled (the VSD group)and 15 patients with fracture (the control group).The plasma miR -155 and miR -222 expression levels were measured by real -time quantitative reverse transcription -polymerase chain reaction (RT -qPCR).The potential target genes of miR -155 and miR -222 were predicted by using 3 current-ly available prediction programs,including TargetScan,mirbase and Miranda,and the signaling pathway of miRNA was predicted by Pathway -express analysis.Results Compared with the control group,the expression levels of miR -155 (P =0.033)and miR -222(P <0.001)in the VSD group decreased significantly;miR -155 and miR -222 predic-ted target genes included 74 and 50,respectively.The Pathway -express analysis indicated that 7 signaling pathways played important roles in the occurrence of fetal VSD,including signaling pathways for heart development,such as:mito-gen -activated protein kinase(MAPK)signaling pathway.Conclusions The expression levels of plasma miR -155 and miR -222 in VSD were significantly decreased.The target genes were related to signaling pathways for heart deve-lopment (MAPK signaling pathway),which indicates that miR -155 and miR -222 may be involved in the pathological process of VSD,and may serve as an independent evaluation indicator for the diagnosis of VSD.

Yishen Jiangzhuo Granules affect tubulointerstitial fibrosis via a mitochondrion-mediated apoptotic pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Yishen Jiangzhuo Granules, YSJZG) on mitochondrial injury and regeneration and renal tubular epithelial cell apoptosis in chronic renal failure (CRF) rats and explore its mechanism from molecular pathology, gene, protein levels, and relative pathway.</p><p><b>METHODS</b>The CRF rat model was established using 5/6 nephrectomy. Sixty rats were randomly divided into six groups: sham-operation group, model (CRF) group, Niaoduqing Granules-treated group [5 g/(kg.day)], low-, moderate-, and high-dose [L-YSJZG, M-YSJZG, H-YSJZG at 3, 6, and 9 g/(kg day)] YSJZG-treated group (n=10 each). The levels of serum creatinine (Scr), blood urea nitrogen (BUN), and 24-h urine protein were assessed after 10 weeks of treatment. The tubulointerstitial injury and collagen deposition were evaluated using periodic acid-schiff stain and Masson staining. Renal tubular epithelial cell apoptosis was assessed using the terminal deoxynucleotidyl transferase dUTP nick end labeling assay, mitochondrial injury was observed using an electron microscope, and superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) levels were assessed using chromometry. Transforming growth factor-β1 (TGF-β1) expression was assessed using immunohistochemistry. The expressions of Bax, Bcl-2, peroxisome proliferator-activated receptor γ coactivator- 1α (PGC-1α), mitochondrial transcription factor A (Tfam), mitogen-activated protein kinases (MAPK) phosphorylation were evaluated by Western blot.</p><p><b>RESULTS</b>YSJZG decreased the 24-h urine protein, BUN, Scr, remnant kidney weight-to-body weight ratio, renal tubular injury, deposition of collagen, and the apoptosis of renal tubular epithelial cells in a dose-dependent manner. YSJZG dose-dependently restored the number and structure of mitochondria and the expression of Tfam and PCG-1α, up-regulated the expression of Bcl-2, and inhibited the expression of Bax. YSJZG also dose-dependently inhibited TGF-β1 expression, increased SOD and GSH activity, decreased the MDA level, and inhibited p38MAPK and pERK1/2 phosphorylation (all P<0.01).</p><p><b>CONCLUSION</b>YSJZG improved the renal function in rats with CRF and inhibited the progression of tubulointerstitial fibrosis by dose-dependently alleviating mitochondrial injury, restoring the expression of Tfam and PCG-1α, and inhibiting renal tubular epithelial cell apoptosis through inhibiting activation of reactive oxygen species-MAPK signaling.</p>

Marsdenia tenacissima extract induces G0/G1 cell cycle arrest in human esophageal carcinoma cells by inhibiting mitogen-activated protein kinase (MAPK) signaling pathway / 中国天然药物

Marsdenia tenacissima extract (MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal cancer and other cancers in the digestive tract. In the present study, the potential mechanism for MTE's activity in esophageal cancer was explored. The effects of MTE on the proliferation of human esophageal cancer cells (KYSE150 and Eca-109) were investigated by the MTT assay, the BrdU (bromodeoxyuridine) incorporation immunofluorescence assay, and flow cytometric analysis. MTE inhibited cell proliferation through inducing G0/G1 cell cycle arrest in KYSE150 and Eca-109. Western blot analysis was employed to determine protein levels in the MTE treated cells. Compared with the control cells, the expression levels of the cell cycle regulatory proteins cyclin D1/D2/D3, cyclin E1, CDK2/4/6 (CDK: cyclin dependent kinase), and p-Rb were decreased significantly in the cells treated with MTE at 40 mg·mL(-1). In addition, MTE had an inhibitory effect on the MAPK (mitogen-activated protein kinase) signal transduction pathway, including ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase), and p38MAPK. Moreover, MTE showed little additional effects on the regulation of cyclin D1/D3, CDK4/6, and p-Rb when the ERK pathway was already inhibited by the specific ERK inhibitor U0126. In conclusion, these data suggest that MTE inhibits human esophageal cancer cell proliferation through regulation of cell cycle regulatory proteins and the MAPK signaling pathways, which is probably mediated by the inhibition of ERK activation.

The effect of apigenin on lipopolysaccharide-induced acute lung injury and its mechanism in mice / 中国中西医结合急救杂志

Objective To observe the effect of apigenin on acute lung injury (ALI) induced by lipopolysaccharide(LPS)in mice,and to discuss its possible mechanism. Methods Forty healthy male Kunming mice were randomly divided using random number table into control group,model group and low,medium,high dose groups of apigenin intervention,and each group consisted of 8 mice. The model of ALI was reproduced by intraperitoneal injection of 5 mg/kg LPS. Mice of the low,medium and high-dose intervention groups were given intraperitoneal injection of apigenin 10,25,50 mg/kg,respectively,1 hour before LPS modeling. Pathological changes in right upper lobe of lung tissue were examined after hematoxylin and eosin(HE)staining and pathology score was observed at 6 hours after modeling. Right inferior lung was weighed to measured wet/dry ratio(W/D). Intercellular adhesion molecule-1(ICAM-1)and tumor necrosis factor-α(TNF-α)in serum and bronchoalveolar lavage fluid (BALF)were determined by enzyme linked immunosorbent assay(ELISA). The mRNA expressions of p38 mitogen-activated protein kinase(p38MAPK),ICAM-1,and TNF-α were determined by reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with control group,lung W/D ratio in model group was significantly increased(17.79±2.89 vs. 5.56±0.37,P<0.05),and the pathology score was significantly elevated(10.32±0.23 vs. 1.87±0.54,P<0.05),ICAM-1 and TNF-α contents,in serum and BALF were increased〔ICAM-1(ng/L) in serum:21.4±2.7 vs. 14.3±3.5,TNF-α(ng/L)in serum:254.8±10.6 vs. 142.3±13.7;ICAM-1(ng/L)in BALF:20.3±2.4 vs. 11.5±3.2,TNF-α(ng/L)in BALF:230.3±5.8 vs. 110.5±11.2,all P<0.05〕,and the mRNA expressions of p38MAPK,ICAM-1 and TNF-α were also increased significantly(the mRNA expression of p38MAPK,ICAM-1 and TNF-αwere 4.42±0.37,4.89±0.27,3.28±0.13,respectively,all P<0.05). Different doses of apigenin could obviously alleviate the damaging effect to the lung,and the most obvious effect was seen in the medium dose group,in which lung W/D ratio was 13.28±1.21,ICAM-1 in serum was(18.5±4.3)ng/L,TNF-αin serum was(169.4±20.8)ng/L,ICAM-1 in BALF was(17.8±3.5)ng/L,TNF-αin BALF was(150.4±7.1)ng/L, the mRNA expression of p38MAPK,ICAM-1 and TNF-αin lung tissue was 2.99±0.28,3.97±0.17,2.87±0.27, respectively. Statistically significant difference was found when they were compared with that of model group(P<0.05 or P<0.01). Conclusion Different doses of apigenin have some antagonistic effect against LPS in producing ALI in mice,the best improvement effect was seen in the medium dose group,and the protective effect may be related to inhibition of p38MAPK signaling pathway activity and reduction of pro-inflammatory factors such as TNF-αand ICAM-1 expression.

Fucoidan induces apoptosis of multiple myeloma cells by inhibiting Ras-p38MAPK and Shh-GLI1 signaling pathways / 肿瘤

Objective: To investigate the effect of fucoidan on multiple myeloma U266 cells and its possible molecular mechanism. Methods: The multiple myeloma U266 cells were treated with 10, 25 and 50 μg/mL fucoidan for 24, 48 and 72 h, respectively. The growth inhibitory rate of U266 cells was determined by cell counting kit-8 (CCK-8) assay. The apoptosis rate was measured by flow cytometry. The expression levels of RAS, p38, phospho-p38, sonic Hedgehog (Shh) and glioma-associated oncogene homolog 1 (GLI1) were detected by Western blotting. Results: The growth of multiple myeloma U266 cells treated with different concentrations of fucoidan (10, 25 and 50 μg/mL) was inhibited only in a dose-dependent manner. The apoptosis rates of U266 cells treated with 10, 25 and 50 μg/mL fucoidan for 24 h were (6.73±2.30)%, (9.12±1.90)% and (20.13±2.10)%, respectively, which were higher than that of the U266 cells without fucoidan intervention [control: (4.08±1.60)%; P < 0.05). The expression levels of RAS, phospho-p38, Shh and GLI1 proteins in U266 cells treated with fucoidan were significantly lower than that of the control (P < 0.05). Conclusion: Fucoidan inhibits proliferation and induces apoptosis of U266 cells by inhibiting Ras-p38MAPK and Shh-GLI1 signaling pathways. Copyright © 2013 by TUMOR.