摘要
目的观察具有抗氧化应激活性的环烯醚萜苷化合物莫诺苷对雷公藤甲素(TP)诱导的肝损伤的保护作用及其分子机制。方法雷公藤甲素,莫诺苷分别单独及联合给药小鼠及人肝癌HepG2细胞后,血清学法测定各组小鼠肝生化指标的变化;MTT法检测各实验组对HepG2细胞生长抑制率的影响;激光共聚焦显微镜观察各组细胞形态的变化;流式细胞术检测各组细胞凋亡率;WB法检测氧化应激通路中关键蛋白的表达。结果动物实验表明雷公藤甲素具有很明显的肝脏损伤作用,小鼠血清中肝生化指标及肝脏指数的水平显著升高(P<0.05);细胞实验则表明其抑制了HepG2细胞的生长,24 h细胞的生长活性仅为72.83%,并随时间增长而降低。细胞凋亡明显,细胞核破裂皱缩,24 h细胞凋亡率高达43.1%。此外,氧化应激通路相关蛋白Nrf2,HO-1的表达明显降低。而莫诺苷联合用药后逆转了以上实验指标,使得小鼠中肝生化指标及肝脏指数显著下降(P<0.05),肝细胞的凋亡率,细胞形态及氧化应激相关通路蛋白的表达都得到了明显改善(P<0.05)。结论莫诺苷可保护雷公藤甲素诱导的肝损伤,其机制可能与抗氧化应激有关。
Objective To investigate the protective effect of monoside against triptolide-induced liver injury and explore its molecular mechanism. Methods BALB/C mice treated with gastric lavage with triptolide and monoside, either alone or in combination, were examined for changes of hepatic biochemical parameters using the serological method. The growth inhibition rate of HepG2 cells treated with triptolide or monoside or both was assessed with MTT assay, and the cell morphological changes were observed using laser confocal microscopy; the expressions of the target proteins in the antioxidative stress pathway were detected using flow cytometry and Western blotting. Results In BALB/C mice, gastric lavage of triptolide induced obvious hepatic damage. In HepG2 cells, treatment with triptolide significantly inhibited the cell growth,resulting in a cell viability as low as 72.83% at 24 h; triptolide also induced obvious cell apoptosis and cell nucleus deformation, causing an apoptosis rate of 43.1% in the cells at 24 h. Triptolide significantly reduced the expressions of Nrf2 and HO-1 proteins related with the oxidative stress pathway. Combined treatment with morroniside obviously reversed these changes, resulting in significantly decreased hepatic biochemical parameters and the liver index in BALB/C mice and in significantly lowered cell apoptosis rate, improved cell morphology, and increased Nrf2 and HO-1 protein expressions in HepG2 cells. Conclusion Monoside protects against triptolide-induced liver injury possibly by relieving oxidative stress.
作者
周玉燕
孙玉
李萍
秦国正
程倩
刘宇
陈滢俐
王国栋
ZHOU Yuyan;SUN Yu;LI Ping;QIN Guozheng;CHENG Qian;LIU Yu;CHEN Yingli;WANG Guodong(Drug Research ＆ Development Center/School of Pharmacy,Wannan Medical College,Wuhu,241002,China;Anhui Provincial Engineering Research Center for Polysaccharide Drugs/Anhui Provincial Key Laboratory of Active Biological Macro-molecules,Wuhu 241002,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2018年第8期949-955,共7页
Journal of Southern Medical University
基金
安徽省自然科学基金(1708085MH207)
安徽省教育厅自然科学基金重点项目(KJ2017A257)
皖南医学院中青年科研基金(WK201609)
皖南医学院大学生科研资助项目(WK2016S23)
皖南医学院博士科研启动基金(WYRCQD201709)
安徽省大学生创新创业计划项目(201610368120
201710368047)
活性生物大分子研究安徽省重点实验室自主研究课题(LAB201801
LAB201804)
