摘要
目的探讨山楂叶总黄酮(HLF)对非酒精性脂肪性肝病(NAFLD)肝细胞凋亡的影响。方法采用肝L-02细胞诱导NAFLD模型,分为正常组、低剂量组、高剂量组、模型组4组,采用不同浓度HLF进行干预,低剂量组HLF100μg/ml,高剂量组HLF400μg/ml。观察细胞的形态学变化,流式细胞术检测细胞凋亡率,We s te rn b lot检测凋亡蛋白表达的变化。结果模型组细胞形态学发生改变,HLF药物干预后,高剂量组细胞形态学改变明显好于低剂量组及模型组;模型组与HLF高剂量组早期凋亡率[(14.63±1.82)%、(6.75±1.83)%]比较,差异有统计学意义(P<0.05);模型组与HLF高剂量组中晚期凋亡率[(11.36±2.94)%、(3.49±1.03)%]比较,差异有统计学意义(P<0.01);HLF高、低剂量组中晚期凋亡率[(3.49±1.03)%、(8.79±1.05)%]比较,差异有统计学意义(P<0.05);模型组与HLF高、低剂量组总凋亡率比较,差异有统计学意义(P<0.05或0.01)。Bax、Cyt-C和Caspase-9促凋亡蛋白在模型组中表达明显增高,Bcl-2、Mitochondria Cytochrome C抗凋亡蛋白在高剂量组及正常组中高表达;模型组与高、低剂量组凋亡蛋白Bax/Bcl-2的比值[(1.999±0.376)、(0.144±0.023)、(0.399±0.045)]比较,差异有统计学意义(P<0.01);模型组与高、低剂量组凋亡蛋白Cytosolic/Mitochondria Cytochrome C的比值[(0.287±0.037)、(0.041±0.006)、(0.097±0.024)]比较,差异有统计学意义(P<0.01);模型组与高、低剂量组凋亡蛋白Cleaved Caspase-9的表达[(0.060±0.005)、(0.030±0.003)、(0.030±0.003)]比较,差异有统计学意义(P<0.01)。结论 NAFLD细胞模型存在凋亡现象,HLF药物可能通过调节Bcl-2、Bax、Cytochrome C、Caspase-9等相关凋亡蛋白的表达,干预肝细胞的凋亡。
Objective To investigate the effects of hawthorn leave flavonoids（HLF）on apoptosis of hepatocytes in non-alcoholic fatty liver disease（NAFLD）.Methods NAFLD model was induced in hepatic L-02 cells,the NAFLD cells were treated with 0,100 or 400？g/ml HLF in model group,low dose HLF group and high dose HLF group,respectively.The morphological changes of cells were observed,the cell apoptosis was detected by flow cytometry,the expression of apoptosis-related proteins was examined by Western Blot.Results Compared to model group,the morphologic changes in HLF-treated groups were attenuated.The early apoptosis rate and middle/late apoptosis rate in high dose HLF group were significantly lower than those in model group（6.75±1.83%vs.14.63±1.82%,P〈0.05;3.49±1.03%vs.11.36±2.94,P〈0.01）.The middle/late apoptosis rate of high dose HLF group was lower than that of low dose HLF group（3.49±1.03%vs.8.79±1.05%,P〈0.05）.The overall apoptosis rate in the model group was significantly higher than that in HLF high and low dose groups,respectively（P〈0.01,P〈0.05）.The expression of apoptosis-related proteins Bax,cytosolic cytochrome C（Cyt-C）and Caspase 9 in the model group increased significantly,while and the expression of anti-apoptosis proteins Bcl-2,Mitochondria Cytochrome C（MCyt-C）was highly expressed in HLF high dose group and normal group.The ratio of Bax/Bcl-2 in the model group was significantly higher than those in HLF high and low dose groups（1.999±0.376 vs.0.144±0.023 and 0.399±0.0454,respectively,P〈0.01）.The ratio of Cytosolic/Mitochondria Cyt-C in the model group was significantly higher than those in HLF high and low dose groups（0.287±0.037 vs.0.041±0.006 and 0.097±0.024 respectively,P〈0.01）.The expression of t cleaved Caspase-9 in the model group was significantly higher than HLF high and low dose groups（0.060±0.005 vs.0.030±0.003 and 0.030±0.003 respectively,P〈0.01）.Conclusion There is obvious cell apoptosis phenomenon in NAFLD,HLF can intervene apopto
作者
黎运呈
曾芳
王秋景
郑迪
杨喆娟
LI Yuncheng;ZENG Fang;WANG Qiujing(Department of Infectious Diseases, Zhoushan Hospital, Zhoushan 316004, China)
出处
《浙江医学》
CAS
2018年第10期1037-1040,1053,1150,共6页
Zhejiang Medical Journal
基金
浙江省中医药科技计划项目(2013ZQ029)
关键词
山楂叶总黄酮
非酒精性脂肪性肝病
凋亡
线粒体
Hawthorn leaves flavonoids Nonalcoholic fatty liver disease Apoptosis Mitochondria
