摘要
目的基于肠-肝轴理论探讨茵陈苓桂剂对高脂饲料诱导的非酒精性脂肪性肝病(NAFLD)大鼠肠黏膜屏障的影响及机制。方法从60只SD大鼠中随机取10只作为对照组,余大鼠采用高脂饲料建立NAFLD模型。造模成功后,将48只大鼠随机分为5组,茵陈苓桂剂高、中、低剂量组分别给予19.6 g/kg、9.8 g/kg、4.9 g/kg的茵陈苓桂剂灌胃,多烯磷脂酰胆碱组给予0.15 g/kg多烯磷脂酰胆碱灌胃,对照组和模型组给予等体积的蒸馏水灌胃。连续灌胃4周后,HE染色观察各组大鼠肝脏及回肠末端组织病理变化,检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、三酰甘油(TG)、总胆固醇(TC)、肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)及血浆内毒素(LPS)水平,Western blot法检测回肠组织中TLR4及CD14蛋白表达情况,免疫组织化学法检测回肠组织中紧密连接蛋白ZO-1、Occludin表达情况。结果与对照组比较,模型组大鼠肝脏和回肠组织可见明显病理损伤,血清ALT、AST、TC、TG、TNF-α、IL-6及血浆LPS水平和回肠组织中TLR4、CD14蛋白相对表达量均明显增高(P均<0.05),回肠组织中ZO-1、Occludin平均光密度值均明显降低(P均<0.05)。与模型组比较,多烯磷脂酰胆碱组与茵陈苓桂剂高、中剂量组大鼠血清ALT、AST、TC、TG水平和茵陈苓桂剂低剂量组大鼠血清AST、TC水平均明显降低(P均<0.05);茵陈苓桂剂高剂量组血清AST水平明显低于中剂量组(P<0.05),血清ALT、TC、TG水平均明显低于低剂量组(P均<0.05)。与模型组比较,茵陈苓桂剂高剂量组大鼠血清TNF-α、IL-6及血浆LPS水平和茵陈苓桂剂中剂量组血清IL-6水平均明显降低(P均<0.05),各给药组大鼠回肠组织中ZO-1、Occludin蛋白阳性表达均增多,TLR4、CD14蛋白相对表达量均明显降低(P均<0.05),茵陈苓桂剂高剂量组大鼠回肠组织中ZO-1、Occludin蛋白表达平均光密度值均明显升高(P均<0.05)。结论茵陈苓桂剂可改善NAFLD大鼠肝脏炎症,调节脂代谢,修复肠道紧密连接,靶向调控LPS/TLR4信号通路的转导,通过调节肠-肝轴、保护肠黏膜屏障功能达到治疗NAFLD的目的。
Objective It is to explore the effect and its mechanism of Yinchen Linggui prescription on intestinal mucosal barrier in rats with non-alcoholic fatty liver disease(NAFLD)induced by high-fat diet based on gut-liver axis.Methods Ten of the 60 SD rats were randomly selected as the control group,and the rest of the rats were used to establish NAFLD models by high-fat diet.After successful modeling,the 48 rats were randomly divided into 5 groups,Yinchen Linggui prescription high,medium and low dose groups were respectively given 19.6 g/kg,9.8 g/kg,4.9 g/kg Yinchen Linggui prescription by gavage,the polyene phosphatidylcholine group was given 0.15 g/kg polyene phosphatidylcholine by gavage,and the control group and model group were given an equal volume of distilled water by gavage.After continuous intragastric administration for 4 weeks,the pathological changes of the liver and terminal ileum were observed by HE staining,and the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),triacylglycerol(TG),total cholesterol(TC),tumor necrosis factor(TNF-α),interleukin-6(IL-6)and plasma endotoxin(LPS)were detected,the expression of TLR4 and CD14 protein in ileum tissue was detected by Western blot method,and the expression of tight junction proteins ZO-1 and Occludin in the ileum tissue were detected by immunohistochemical method.Results Compared with the control group,the liver and ileum tissue of the model group showed obvious pathological damage,the levels of serum ALT,AST,TC,TG,TNF-α,IL-6 and plasma LPS and the relative expression of TLR4 and CD14 protein in the ileum tissue were all significantly increased(P<0.05),the average optical density of ZO-1 and Occludin in the ileum tissue was significantly decreased(both P<0.05).Compared with the model group,the levels of serum ALT,AST,TC,TG of rats in the polyene phosphatidylcholine group and the Yinchen Linggui prescription high and medium dose groups and the levels of serum AST and TC in the Yinchenlinggui prescription low dose group were significantly decreased(all P<0.05);the level of serum AST in the Yinchen Linggui prescription high-dose group was significantly lower than that in the middle-dose group(P<0.05),and the levels of serum ALT,TC,and TG in the high dose group were significantly lower than those in the low-dose group(P<0.05).Compared with the model group,the levels of serum TNF-α,IL-6 and plasma LPS in the Yinchenlinggui prescription high-dose group and the serum IL-6 levels of the Yinchenlinggui prescription middle-dose group were significantly reduced(all P<0.05).The positive expression of ZO-1 and Occludin protein in the ileum tissue of rats in the administration group was increased,while the relative expression of TLR4 and CD14 protein was significantly reduced(all P<0.05).The average optical density value of ZO-1 and Occludin protein expression in the ileum tissue of the high-dose Yinchenlinggui prescription group was increased significantly(all P<0.05).Conclusion Yinchen Linggui prescription can improve liver inflammation,regulate lipid metabolism,repair intestinal tight junctions in NAFLD rats,regulate LPS/TLR4 signaling pathway,and achieve treatment of NAFLD by regulating gut-liver axis and protecting intestinal mucosal barrier function.
作者
吴迪
谢春娥
李峰
王允亮
薛晓轩
袁亚利
王乾皓
焦瑶
李军祥
杨晋翔
WU Di;XIE Chun’e;LI Feng;WANG Yunliang;XUE Xiaoxuan;YUAN Yali;WANG Qianhao;JIAO Yao;LI Junxiang;YANG Jinxiang(Beijing University of Chinese Medicine,Beijing 100029,China;The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China;Dongfang Hospital of Beijing University of Chinese Medicine,Beijing 100078,China)
出处
《现代中西医结合杂志》
CAS
2021年第6期571-577,共7页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
北京中医药大学科研创新团队项目(2019-JYB-TD004)
北京中医药大学自主选题(2019-JYB-XS-214)。
关键词
非酒精性脂肪性肝病
茵陈苓桂剂
肠黏膜屏障
肠-肝轴
肝病实脾
non-alcoholic fatty liver disease
Yinchen Linggui prescription
intestinal mucosal barrier
gut-liver axis
treating liver by nourishing spleen
