| 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HLF100滋g/ml，高剂量组HLF400滋g/ml。观察细胞的形态学变化，流式细胞术检测细胞凋亡率，Westernblot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、MitochondriaCytochromeC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/MitochondriaCytochromeC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CleavedCaspase-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、CytochromeC、Caspase-9等相关凋亡蛋白的表达，干预肝细胞的凋亡。 |
| 关键词: 山楂叶总黄酮 非酒精性脂肪性肝病 凋亡 线粒体 |
| DOI：10.12056/j.issn.1006-2785.2018.40.10.2018-101 |
| 分类号: |
| 基金项目:浙江省中医药科技计划项目（2013ZQ029） |
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| Effect of hawthorn leave flavonoids on apoptosis of hepatocyts in non-alcoholic fatty liver disease |
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Zhoushan Hospital
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| Abstract: |
| 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 Caspase9 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 apoptosis of hepatocytes through regulating the expression of Bcl-2, Bax, Cytochrome C, Caspase-9 and other apoptosis-re- lated proteins. |
| Key words: Hawthorn leaves flavonoids Nonalcoholic fatty liver disease Apoptosis Mitochondria |
