Objective: To investigate the mechanism of inflammatory-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 ...Objective: To investigate the mechanism of inflammatory-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(柴胡疏肝散, CHSG) group(3.2 g/kg), high-dose CHSG group(9.6 g/kg), low-dose Shenling Baizhu Powder(参苓白术散, 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 fluorescence 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(P<0.05 or P<0.01), while those indices were significantly ameliorated in the H-IR group(P<0.05 or P<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(P<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(P<0.05 or P<0.01). The m RNA 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(P<0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group(P<0.05 or P<0.01). Conclusions: Inflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38 MAPK signaling pathway in KCs for the anti-inflammatory effect of soothing Gan and invigorating Pi recipes.展开更多
Objective To investigate the mechanism of lipid metabolism disorders in Kupffer cells （KCs） of non-alcoholic fatty liver disease （NAFLD） rats mediated by LXRα-SREBP-lc pathway and the interference of soothing liv...Objective To investigate the mechanism of lipid metabolism disorders in Kupffer cells （KCs） of non-alcoholic fatty liver disease （NAFLD） rats mediated by LXRα-SREBP-lc pathway and the interference of soothing liver and invigorating spleen recipe （SLISR） on it. Methods SD male rats were randomly divided into five groups： normal, model, soothing liver recipe （SLR）, invigorating spleen recipe （ISR）, and soothing liver and invigorating spleen recipe （SLISR） groups. The rats in treatment groups were administered for 8 weeks. The liver tissue was stained with H＆E and oil red O. The levels of hepatic lipid and blood lipid were measured by biochemical analyzer. KCs were isolated from the livers of rats to evaluate the expression of LXRα, SREBP-1 C, and FAS mRNA by real-time fluorescence quantitative PCR tests; LXRα, SREBP-1C, and FAS proteins were measured by Western blotting. Results The H＆E and oil red O staining results showed that the model rats successfully reproduced typical pathogenetic and histopathological features of NAFLD. The levels of hepatic lipid and blood lipid in the model rats were dramatically increased. Compared with the model group, the values of hepatic lipid and blood lipid in the treatment groups were significantly ameliorated （P〈 0.05, 0.01 ）. The yields of purified KCs from each rat were 2×10^7-3×10^7. The viability of KCs was higher than 95%, with the purity over 90.18%. Compared with the model group, the expression of LXRα, SREBP-1C, and FAS mRNA and proteins was decreased in all treatment groups, especially in the SLR group （P〈 0.05）. Conclusion SLISR may protect liver against injury included by lipid metabolism disorders in KCs through LXRα/ SREBP-1 c signaling pathway, which may be an important mechanism for the prevention and treatment of NAFLD.展开更多
目的:运用网络药理学和分子对接的方法,探索健脾疏肝方治疗非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的潜在机制。方法:利用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology,TC...目的:运用网络药理学和分子对接的方法,探索健脾疏肝方治疗非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的潜在机制。方法:利用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology,TCMSP)和已发表的文献筛选健脾疏肝方的活性化合物和潜在作用靶点,利用人类基因数据库(GeneCards)、在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)和药物靶标数据库(therapeutic target database,TTD)筛选NAFLD的相关靶点,预测健脾疏肝方治疗NAFLD的潜在靶点。利用STRING数据库和Cytoscape软件构建共同靶标的PPI网络。利用DAVID数据库及微生信云平台进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。最后利用AutoDock Vina和Pymol软件对核心靶点和主要活性化合物进行分子对接。结果:共筛选出117个活性化合物和279个潜在靶标,其中槲皮素(Quercetin)、木犀草素(luteolin)、豆甾醇(Stigmasterol)、山柰酚(kaempferol)为健脾疏肝方主要活性化合物。PPI网络显示白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、信号转换器和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、前列腺素内过氧化物合酶2(prostaglandin-endoperoxide synthase 2,PTGS2)和血管内皮生长因子A(vascular endothelial growth factor,VEGFA)是关键靶蛋白。KEGG通路富集分析结果显示,癌症、TNF、乙型病毒性肝炎、MAPK、PI3K-Akt及NAFLD信号通路可能是健脾疏肝方干预NAFLD的潜在机制。分子对接结果表明,健脾疏肝方主要活性化合物与核心靶标具有良好的结合能力。结论:健脾疏肝方可以通过多组分、多靶点及多通路起到干预NAFLD的作用。展开更多
基金Supported by the National Natural Science Foundation of China(No.30973694)
文摘Objective: To investigate the mechanism of inflammatory-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(柴胡疏肝散, CHSG) group(3.2 g/kg), high-dose CHSG group(9.6 g/kg), low-dose Shenling Baizhu Powder(参苓白术散, 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 fluorescence 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(P<0.05 or P<0.01), while those indices were significantly ameliorated in the H-IR group(P<0.05 or P<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(P<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(P<0.05 or P<0.01). The m RNA 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(P<0.01), while those expression levels in the L-IR and H-IR groups were significantly lower than the model group(P<0.05 or P<0.01). Conclusions: Inflammation in KCs might play an important role in the pathogenesis of NASH in rats. The data demonstrated the importance of TLR4-p38 MAPK signaling pathway in KCs for the anti-inflammatory effect of soothing Gan and invigorating Pi recipes.
基金National Natural Science Foundation of China (81173216)Foundation for Scientific Research Fostering and Innovation of Jinan University (21612118)
文摘Objective To investigate the mechanism of lipid metabolism disorders in Kupffer cells （KCs） of non-alcoholic fatty liver disease （NAFLD） rats mediated by LXRα-SREBP-lc pathway and the interference of soothing liver and invigorating spleen recipe （SLISR） on it. Methods SD male rats were randomly divided into five groups： normal, model, soothing liver recipe （SLR）, invigorating spleen recipe （ISR）, and soothing liver and invigorating spleen recipe （SLISR） groups. The rats in treatment groups were administered for 8 weeks. The liver tissue was stained with H＆E and oil red O. The levels of hepatic lipid and blood lipid were measured by biochemical analyzer. KCs were isolated from the livers of rats to evaluate the expression of LXRα, SREBP-1 C, and FAS mRNA by real-time fluorescence quantitative PCR tests; LXRα, SREBP-1C, and FAS proteins were measured by Western blotting. Results The H＆E and oil red O staining results showed that the model rats successfully reproduced typical pathogenetic and histopathological features of NAFLD. The levels of hepatic lipid and blood lipid in the model rats were dramatically increased. Compared with the model group, the values of hepatic lipid and blood lipid in the treatment groups were significantly ameliorated （P〈 0.05, 0.01 ）. The yields of purified KCs from each rat were 2×10^7-3×10^7. The viability of KCs was higher than 95%, with the purity over 90.18%. Compared with the model group, the expression of LXRα, SREBP-1C, and FAS mRNA and proteins was decreased in all treatment groups, especially in the SLR group （P〈 0.05）. Conclusion SLISR may protect liver against injury included by lipid metabolism disorders in KCs through LXRα/ SREBP-1 c signaling pathway, which may be an important mechanism for the prevention and treatment of NAFLD.
文摘目的:运用网络药理学和分子对接的方法,探索健脾疏肝方治疗非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的潜在机制。方法:利用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology,TCMSP)和已发表的文献筛选健脾疏肝方的活性化合物和潜在作用靶点,利用人类基因数据库(GeneCards)、在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)和药物靶标数据库(therapeutic target database,TTD)筛选NAFLD的相关靶点,预测健脾疏肝方治疗NAFLD的潜在靶点。利用STRING数据库和Cytoscape软件构建共同靶标的PPI网络。利用DAVID数据库及微生信云平台进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。最后利用AutoDock Vina和Pymol软件对核心靶点和主要活性化合物进行分子对接。结果:共筛选出117个活性化合物和279个潜在靶标,其中槲皮素(Quercetin)、木犀草素(luteolin)、豆甾醇(Stigmasterol)、山柰酚(kaempferol)为健脾疏肝方主要活性化合物。PPI网络显示白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、信号转换器和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、前列腺素内过氧化物合酶2(prostaglandin-endoperoxide synthase 2,PTGS2)和血管内皮生长因子A(vascular endothelial growth factor,VEGFA)是关键靶蛋白。KEGG通路富集分析结果显示,癌症、TNF、乙型病毒性肝炎、MAPK、PI3K-Akt及NAFLD信号通路可能是健脾疏肝方干预NAFLD的潜在机制。分子对接结果表明,健脾疏肝方主要活性化合物与核心靶标具有良好的结合能力。结论:健脾疏肝方可以通过多组分、多靶点及多通路起到干预NAFLD的作用。
