活血理肝汤通过LPS/TLR-4/NF-kB信号通路治疗NAFLD大鼠的机制探讨  

Mechanism of Huoxue Ligan Decoction(活血理肝汤)in Preventing and Treating Nafld Through Lps/TIr4/Nf-Kb Signal Pathway

作　　者：刘星 陶竹 吕呈 张彤彤 张朕 王岩[1] LIU Xing;TAO Zhu;LYU Cheng;ZHANG Tongong;ZHANG Zhen;WANG Yan(The Sirth People Hospital of Shenyang,Shenyang Hepathobiliary Clinical Medicine Research Center,Shenyang 110006,Liaoning,China)

机构地区：[1]沈阳市第六人民医院,沈阳市肝胆临床医学研究中心,辽宁沈阳110006

出　　处：《实用中医内科杂志》2023年第9期42-45,I0007,I0008,共6页Journal of Practical Traditional Chinese Internal Medicine

基　　金：中国公共卫生联盟课题项目(GWLM202009)。

摘　　要：目的 探讨活血理肝汤通过LPS/TLR-4/NF-kB信号通路防治NAFLD的机制。方法 30只SD大鼠随机分为正常对照组、模型组和活血理肝汤组,每组10只。正常对照组给予普通饲料喂养;模型组和活血理肝汤组给予高脂饲料喂养。8周后经肝脏病理HE染色确认造模成功,活血理肝汤组按照8.5g/(kg·d)进行灌胃,正常对照组和模型组使用相同体积生理盐水灌胃,持续8周。全自动生化分析仪测定各组大鼠血清血脂、肝功水平,HE染色和油红O染色观察各组大鼠肝脏组织形态与脂质沉积情况;酶联免疫吸附法检测各组大鼠肝脏白细胞介素-1(IL-1)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)水平以及血清、肝脏、盲肠内容物的脂多糖(LPS)水平。Western blot法和荧光定量PCR法分别检测各组大鼠肝脏组织NF-κB和TLR4的蛋白和mRNA表达水平。结果 与正常对照组比较,模型组大鼠血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)和天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)水平显著升高,高密度脂蛋白(HDL-C)水平显著降低(P<0.05);肝细胞内可见大量大小不等的脂肪空泡,肝窦狭窄或消失,脂质沉积明显;肝脏IL-1、IL-6、IL-8、TNF-α水平以及血清、肝脏、盲肠内容物中LPS水平均显著升高(P<0.05);肝脏TLR4和NF-κB的蛋白和mRNA表达均增高(P<0.05)。与模型组比较,活血理肝汤组大鼠血清TC、TG、LDL-C和AST、ALT水平显著降低(P<0.05),HDL-C水平升高(P<0.05);肝细胞内脂肪空泡明显减少,肝窦狭窄有所缓解,脂质沉积情况有所改善;肝脏IL-1、IL-6、IL-8、TNF-α水平和盲肠内容物、血清、肝脏LPS水平均显著降低(P<0.05);肝脏TLR4和NF-κB的蛋白和mRNA的表达均降低(P<0.05)。结论 活血理肝汤能够改善NAFLD大鼠血脂水平、减轻肝脏脂质沉积,降低炎症指标IL-1、IL-6、IL-8和TNF-α的水平,改善肝脏的内环境,并可能是通过LPS/TLR-4/NF-kB信�Objective To Explore the possible mechanism of Huoxue Ligan Decoction(活血理肝汤)in preventing and treating NAFLD through LPS/TLR4/NF-kB signal pathway.Methods Thirty SD rats were randomLy divided into control group(10 rats),model group(10 rats)and Huoxue Ligan Decoction group(10 rats).The control group was fed with normal diet,and the model group and Huoxue Ligan Decoction group were fed with high-fat diet.After 8 weeks,the group of Huoxue Ligan Decoc-tion was given 8.5 g/(kg·d)by gavage,and the control group and model group were given the same volume of normal saline by gavage for 8 weeks.The automatic biochemical analyzer was used to detect serum lipids and liver function of each group;The HE staining and Oil Red O staining were used to observe the liver morphology and lipid deposition.Enzyme linked immunosorbent assay was used to detect interleukin-1(IL-1),interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor-alpha(TNF-α)and lipopolysaccharide(LPS)levels in serum,liver and cecal contents.Western blot and RT-qPCR were used to detect the Protein and mRNA expression levels of NF-kB and TLR4 in liver tissue.Results Compared with the control group,the levels of serum triglyceride(TG),total cholesterol(TC),low density lipoprotein(LDL-C),aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in the model group were significantly increased,while the levels of high density lipoprotein(HDL-C)were significantly decreased(P<0.05).The liver morphology exhibited massive fat vacuole,stenotic or disappeared hepatic sinuses and obvious fatty deposition.The IL-1,IL-6,IL-8,TNF-αlevel in liver and the LPS level in serum,liver and cecal contents were significantly increased,as well as the protein and mRNA expression level of TLR4 and NF-kB(P<O.05).Compared with the model group,the serum TC,TG,LDL-C,AST and ALT levels in the Huo-Xue-Li-Gan Decoction group decreased significantly(P<0.05),while the HDL-C level increased(P<0.05).The liver morphology showed the stenosis of hepatic sinuses was relieved,the fatt

关 键 词：活血理肝汤 非酒精性脂肪性肝病 LPS/TLR4/NF-κB通路 

分 类 号：R285.5[医药卫生—中药学]