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 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.

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.