摘要
目的明确龙胆苦甙对大鼠非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)的疗效,并初步探讨LC3Ⅱ基因及自噬在其中所起的作用及机制。方法建立SD大鼠NASH模型,分为:正常对照组、模型对照组、龙胆苦甙组、龙胆苦甙+3-MA组、二甲双胍阳性对照组,每组8只大鼠。给予相应治疗和处理4周后,通过q-PCR及Western blot分别检测大鼠肝组织LC3Ⅱ、LC3Ⅰ的mRNA和蛋白质表达,HE染色检测肝组织病理学情况,结合大鼠血清的血脂(CHOL和TG)、谷丙转氨酶以及炎症因子(IL-1β和TNF-α)水平的变化,评价龙胆苦甙对大鼠NASH的治疗效果,并初步探讨LC3Ⅱ及自噬在这一过程中所起的作用和机制。结果(1)正常对照组肝小叶结构正常,未见脂肪变及炎症损伤;模型对照组肝细胞索结构紊乱,出现严重肝脂肪变及炎症损伤,并可见点、片状坏死形成;龙胆苦甙组、二甲双胍阳性对照组肝脂肪变及炎症损伤均有所减轻;龙胆苦甙+3-MA组肝脂肪变和炎症损伤较龙胆苦甙组明显加重,肝小叶结构紊乱,坏死增多。(2)模型对照组ALT、CHOL、TG、IL-1β、TNF-α水平均较正常对照组升高,龙胆苦甙组、二甲双胍阳性对照组的ALT、CHOL、TG、IL-1β、TNF-α水平一定程度上得到逆转,而龙胆苦甙+3-MA组ALT、CHOL、TG、IL-1β、TNF-α水平又较龙胆苦甙组升高,差异有统计学意义(P<0.05)。(3)模型对照组大鼠肝组织LC3ⅡmRNA和蛋白质表达较正常对照组明显下调,这一趋势在龙胆苦甙组和二甲双胍阳性对照组中出现逆转,而龙胆苦甙+3-MA组的LC3ⅡmRNA和蛋白质表达又明显下调,差异有统计学意义(均P<0.05)。结论龙胆苦甙可缓解大鼠NASH相关肝损伤,其机制可能与龙胆苦甙上调肝组织LC3Ⅱ表达从而促进了自噬有关。
Objective To clarify the effect of gentiopicroside on alleviating nonalcoholic steatohepatitis(NASH)in rats,and to explore the role of LC3II gene and autophagy in this process.Methods Establishment of SD rat NASH model was established and assigned to normal control group,model control group,gentiopicroside group,gentiopicroside+autophagy inhibitor 3-methyladenine group,metformin positive control group,8 rats in each group.After 4 weeks of treatment,mRNA and protein expression of LC3Ⅱwere detected,LC3Ⅰin liver tissue was assessed by q-PCR and Western blot,HE staining was performed for liver histopathology,the therapeu⁃tic effect of gentiopicroside on NASH in rats was evaluated with serum lipids(CHOL and TG),alanine aminotrans⁃ferase and inflammatory factor(IL-1βand TNF-α)changes.Results(1)In the normal control group,the structure of hepatic lobule was normal,without fatty change and inflammatory damage;In the model control group,the structure of hepatocyte cord was disordered,with severe liver steatosis and inflammatory damage,and spot and flaky necrosis was observed;The liver steatosis and inflammatory damage were alleviated in gentiopicroside group and metformin positive control group;Compared with gentiopicroside group,liver steatosis and inflammatory damage in gentiopicroside+3⁃MA group were significantly aggravated,and the structure of hepatic lobule was disordered and necrosis was increased.(2)The ALT,CHOL,TG,IL-1β,TNF-αlevels of NASH model control group were significantly higher than those in the normal control group,the ALT,CHOL,TG,IL-1β,TNF-αlevels of gentio⁃picroside group,metformin positive control group were reversed to some extent,while gentiopicroside+3⁃MA group the level of ALT,CHOL,TG,IL-1β,TNF-αis higher than that of gentiopicroside group,the difference was statistically significant,all P<0.05.(3)The expression of LC3ⅡmRNA and protein in the liver tissue of rats in the model control group was significantly lower than that in the normal control group.This trend was reversed in the gentiopicroside group and the metformin positive control group,while the expression of LC3ⅡmRNA and protein in the gentiopicroside+3-MA group was significantly lower,the difference was statistically significant,all P<0.05.Conclusion Gentiopicroside could alleviate NASH-related liver injury in rats.Its mechanism may be related to GPS up-regulating the expression of LC3Ⅱin liver tissue and promoting autophagy.
作者
丁洁
刘思奇
王艺颖
王霖
施承宏
王芳
常国楫
华丽娟
陈华憶
李生浩
王晴晴
DING Jie;LIU Siqi;WANG Yiying;WANG Lin;SHI Chenghong;WANG Fang;CHANG Guoji;HUA Lijuan;CHEN Huayi;LI Shenghao;WANG Qingqing(Department of Hepatopathy,the 3rd People′s Hospital of Kunming/Clinical-Medical Center of Infectious Disease of Yunnan Province,Kunming 650041,China;不详)
出处
《实用医学杂志》
CAS
北大核心
2023年第16期2022-2028,共7页
The Journal of Practical Medicine
基金
国家自然科学基金地区科学基金项目(编号:82260408)
云南省科技厅基础研究专项面上项目(编号:202101AT070054)
昆明市卫健委卫生科研课题(编号:2022-03-08-005,2022-03-08-011,2021-03-08-005)。
关键词
龙胆苦甙
微管相关蛋白1轻链3Ⅱ
微管相关蛋白1轻链3Ⅰ
自噬
非酒精性脂肪性肝炎
gentiopicroside
microtubule associated protein 1 light chain 3Ⅱ
microtubule associated protein 1 light chain 3Ⅰ
autophagy
non-alcoholic steatohepatitis
