基于LPS/TLR4通路研究疏肝理脾汤治疗NASH大鼠的作用机制
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摘要
目的:探讨疏肝理脾汤对非酒精性脂肪性肝炎(NASH)大鼠脂多糖-Toll受样体-4(LPS-TLR4)信号通路的影响。方法:造模成功的NASH 84只大鼠随机分为6组,其中两组为无中药干预对照组,分为蛋氨酸胆碱缺乏(MCD)饮食(M_M)组和正常饮食(M_N)组;两组继续MCD饮食加中药(疏肝理脾汤)灌服,分以中药低剂量(D_低)组、高剂量(D_高)组;两组为正常饮食加中药(疏肝理脾汤)灌服,分以中药低剂量(Z_低)组、高剂量(Z_高)组,持续4周,于实验9周末处死所有动物,并行大鼠肝脏组织病理、外周血相关生化指标、细胞相关炎症因子、血浆LPS水平、肝组织TLR4水平等相关检测与分析比较。结果:与空白组比较,实验组SAF积分,血清ALT、AST、TC、TG、TNF-α、IL-6、IL-1含量;血浆LPS、TLR4mRNA及蛋白水平表达均明显升高;实验组内比较,与两组模型组(M_M组、M_N组)比较,各治疗组(Z_高组_、Z_低组、D_低组、D_高组)SAF积分;血清ALT、AST、TC、TG、TNF-α、IL-6、IL-1含量;血浆LPS、TLR4mRNA及蛋白水平表达均显著降低(P<0.05)。各治疗组内两两比较,Z_高组在改善SAF积分、降低血清ALT、AST、TC、TG、TNF-α、IL-6、IL-1含量、降低血浆LPS/TLR4mRNA及蛋白水平表达方面优于其他治疗组(P<0.05)。结论:"疏肝理脾汤"高剂量联合正常饮食对改善NASH大鼠脂肪沉积,降低炎症状态有一定作用,干预机制可能与影响LPS/TLR4信号通路,从而达到抑制相关炎症细胞因子分泌,减轻肝细胞炎性反应及脂肪变性有关。
Objective:To discuss the effect of Shugan Lipi Decoction Ameliorates non-alcoholic steatohepatitis in SD rats through regulation of the LPS/TLR4 signaling pathway.Methods:Total of 84 successful NASH rats were randomly divided into 6 groups.The two groups were divided into MCD diet(MM) group and normal diet(MN) group.The two groups were fed with MCD diet plus Chinese medicine(Shugan Lipi Decoction low dose and Shugan Lipi Decoction high dose).The two groups were fed with normal diet plus traditional Chinese medicine(Shugan Lipi Decoction low dose and Shugan Lipi Decoction high dose), after 4 weeks, all animals were killed at the end of the 9 th week of the Liver histopathology, peripheral blood biochemical parameters, cell-related inflammatory factors, plasma LPS level and liver tissue TLR4 level in rats were analyed.Results:Compared with black group, SAF scores,ALT,AST,TC,TG, TNF-α,IL-6,IL-1 in serum,LPS in plasma and the expression of protein and mRNA of TLR4 of Experience groups increased obviously(P<0.05).Compared with model groups(MMgroup、MN group),SAF scores,ALT,AST,TC,TG,TNF-α,IL-6,IL-1 in serum,LPS in plasma and the expression of protein and mRNA of TLR4 reduced in treatment group decreased(P<0.05),Comparison between each treatment group, Z higah group was superior to other treatment groups in improving SAF score, reducing of ALT, AST, TC, TG, TNFα, IL-6, IL-1 content in serum, reducing LPS in plasma, and reducingTLR4 mRNA and protein expression(P<0.05).Conclusion:high dose of "Shugan Lipi Decoction" combined with normal diet can ameliorate fat deposition and reduce inflammation in NASH rats. Intervention mechanism may be related to the influence of LPS/TLR4 signaling pathway, so as to inhibit the secretion of related inflammatory cytokines, alleviate hepatocyte inflammatory response and steatosis.
Objective:To discuss the effect of Shugan Lipi Decoction Ameliorates non-alcoholic steatohepatitis in SD rats through regulation of the LPS/TLR4 signaling pathway.Methods:Total of 84 successful NASH rats were randomly divided into 6 groups.The two groups were divided into MCD diet(MM) group and normal diet(MN) group.The two groups were fed with MCD diet plus Chinese medicine(Shugan Lipi Decoction low dose and Shugan Lipi Decoction high dose).The two groups were fed with normal diet plus traditional Chinese medicine(Shugan Lipi Decoction low dose and Shugan Lipi Decoction high dose), after 4 weeks, all animals were killed at the end of the 9 th week of the Liver histopathology, peripheral blood biochemical parameters, cell-related inflammatory factors, plasma LPS level and liver tissue TLR4 level in rats were analyed.Results:Compared with black group, SAF scores,ALT,AST,TC,TG, TNF-α,IL-6,IL-1 in serum,LPS in plasma and the expression of protein and mRNA of TLR4 of Experience groups increased obviously(P<0.05).Compared with model groups(MMgroup、MN group),SAF scores,ALT,AST,TC,TG,TNF-α,IL-6,IL-1 in serum,LPS in plasma and the expression of protein and mRNA of TLR4 reduced in treatment group decreased(P<0.05),Comparison between each treatment group, Z higah group was superior to other treatment groups in improving SAF score, reducing of ALT, AST, TC, TG, TNFα, IL-6, IL-1 content in serum, reducing LPS in plasma, and reducingTLR4 mRNA and protein expression(P<0.05).Conclusion:high dose of "Shugan Lipi Decoction" combined with normal diet can ameliorate fat deposition and reduce inflammation in NASH rats. Intervention mechanism may be related to the influence of LPS/TLR4 signaling pathway, so as to inhibit the secretion of related inflammatory cytokines, alleviate hepatocyte inflammatory response and steatosis.
引文
[1] 中华医学会肝病学分会脂肪肝和酒精性肝病学组,中国医师协会脂肪性肝病专家委员会,范建高等.非酒精性脂肪性肝病防治指南(2018年更新版)[J].实用肝脏病杂志,2018,21(2):177-186.
[2] Jiang JW,Chen XH,Ren Z,et al.Gut microbial dysbiosis associates hepato-cellular carcinoma via the gut-liver axis[J].Hepatob Pancreat Dis Int,2019,18(1):19-27.
[3] 卢胜家,熊焰.超微脂康饮对非酒精性脂肪性肝病大鼠肝脏COX-2表达的影响[J].中国肝脏病杂志,2012,17(7):498-500.
[4] 董婷,卢胜家,熊焰.氧化应激在非酒精性脂肪性肝炎发病机制中的作用研究进展[J].中国肝脏病杂志,2012,18(8):602-604.
[5] Bedossa P.Utility and appropriateness of the fatty liver inhibition of progression (FLIP) algorithm and steatosis,activity,and fibrosis (SAF) score in the evaluation of biopsies of nonalcoholic fatty liver disease[J].Hepatology,2014,60(2)565-575.
[6] 李志国,姜韩雪,焦云涛,等.非酒精性脂肪性肝病中医证型分布的文献分析[J].中华中医药杂志,2017,32(10):4713-4715.
[7] Rocha DM,Caldas AP,Oliveira L,et al.Saturated fatty acids trigger TLR4-mediated inflammatory response.[J].Atherosclerosis,2016,46(1):211-215.
[8] Plóciennikowska A,Hromada-Judycka A,Borzecka K,et al.Cooperation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling[J].Cell Mol Life Sci,2015,72(3):557-581.
[9] Anna A,Guido C,Oliveira FL,et al.The Role of Tissue Macrophage-Mediated Inflammation on NAFLD Pathogenesis and Its Clinical Implications[J].Mediat Inflammat,2017,26(1):1-15.
[10] Sutter AG,Palanisamy AP,Lench JH,et al.Dietary Saturated Fat Promotes Development of Hepatic Inflammation Through Toll-Like Receptor 4 in Mice[J].J Cell Biochem,2016,117(7):1613-1621.
[11] Vespasiani-Gentilucci U,Gallo P,Picardi A.The role of intestinal microbiota in the pathogenesis of NAFLD:starting points for intervention[J].Arch Med Sci,2018,14(3):701-706.
[2] Jiang JW,Chen XH,Ren Z,et al.Gut microbial dysbiosis associates hepato-cellular carcinoma via the gut-liver axis[J].Hepatob Pancreat Dis Int,2019,18(1):19-27.
[3] 卢胜家,熊焰.超微脂康饮对非酒精性脂肪性肝病大鼠肝脏COX-2表达的影响[J].中国肝脏病杂志,2012,17(7):498-500.
[4] 董婷,卢胜家,熊焰.氧化应激在非酒精性脂肪性肝炎发病机制中的作用研究进展[J].中国肝脏病杂志,2012,18(8):602-604.
[5] Bedossa P.Utility and appropriateness of the fatty liver inhibition of progression (FLIP) algorithm and steatosis,activity,and fibrosis (SAF) score in the evaluation of biopsies of nonalcoholic fatty liver disease[J].Hepatology,2014,60(2)565-575.
[6] 李志国,姜韩雪,焦云涛,等.非酒精性脂肪性肝病中医证型分布的文献分析[J].中华中医药杂志,2017,32(10):4713-4715.
[7] Rocha DM,Caldas AP,Oliveira L,et al.Saturated fatty acids trigger TLR4-mediated inflammatory response.[J].Atherosclerosis,2016,46(1):211-215.
[8] Plóciennikowska A,Hromada-Judycka A,Borzecka K,et al.Cooperation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling[J].Cell Mol Life Sci,2015,72(3):557-581.
[9] Anna A,Guido C,Oliveira FL,et al.The Role of Tissue Macrophage-Mediated Inflammation on NAFLD Pathogenesis and Its Clinical Implications[J].Mediat Inflammat,2017,26(1):1-15.
[10] Sutter AG,Palanisamy AP,Lench JH,et al.Dietary Saturated Fat Promotes Development of Hepatic Inflammation Through Toll-Like Receptor 4 in Mice[J].J Cell Biochem,2016,117(7):1613-1621.
[11] Vespasiani-Gentilucci U,Gallo P,Picardi A.The role of intestinal microbiota in the pathogenesis of NAFLD:starting points for intervention[J].Arch Med Sci,2018,14(3):701-706.