| 摘要: |
| 目的:探索安石榴甙(PUN)在机械力诱导成纤维细胞凋亡中的作用及其可能机制。方法:不同浓度(0、2.5、5、10、25、50、100μmol/L)的PUN处理L929细胞不同时间(12、24、48小时)后,采用CCK8法检测细胞增殖活性;采用Western blot检测不同浓度PUN(0、2.5、5、10、25μmol/L)处理24小时后Nrf2蛋白表达;将L929细胞根据是否行PUN(10μmol/L,24小时)预处理和是否采用四点弯曲力学加载系统进行机械力干预(5333μstrain,1 Hz,4小时)分为4组:空白对照组、加力组、PUN组及PUN加力组,采用流式细胞术检测各组细胞凋亡率;采用Western blot检测凋亡相关蛋白Bax、Bcl-2及caspase3的表达;采用H2DCF-DA荧光探针检测各组细胞内活性氧簇(ROS)水平的变化。结果:PUN在浓度10μmol/L、作用时间24小时对细胞增殖活性及Nrf2蛋白表达的促进作用最显著;与对照组相比,加力组细胞凋亡率明显增加、ROS水平显著上升、Bax表达和Active-caspase3/caspase3比率明显增加(P<0.05);与加力组相比,PUN加力组细胞凋亡率显著减少、ROS水平明显降低、Bcl2表达显著增加、Bax表达和Active-caspase3/caspase3比率降低(P<0.05)。结论:PUN可明显减少机械力诱导的成纤维细胞凋亡,其机制可能是通过上调抗氧化蛋白Nrf2的表达,从而抑制机械力诱导ROS水平上升所导致的氧化损伤。 |
| 关键词: 安石榴甙 机械力 压力性尿失禁 氧化应激 凋亡 |
| DOI： |
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| 基金项目:国家自然科学基金面上项目(编号:81471442,81270684) |
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| Study on the Effect and Mechanism of Punicalagin on Mechanical Stress Induced Apoptosis in Fibroblasts |
| WANG Linlin;LI Bingshu;LIU Cheng;HONG Li |
| (Renmin Hospital of Wuhan University) |
| Abstract: |
| Objective: To investigate the effect of PUN( Punicalagin) on the apoptosis of fibroblasts induced by mechanical injury and its possible mechanism. Methods: We treated L929 cells with different concentrations( 0,2. 5,5,10,25,50,100 μmol/L) of PUN at different time( 12,24,48 h). CCK8 method was used to detect cell proliferation activity. Western blot was used detect different concentrations of PUN( 0,2. 5,5,10,25 μmol/L) with expression of Nrf2 protein in L929 cells after 24 h. According to whether or not pretreatment of PUN( 10 μmol/L,24h) and the four point bending loading system for mechanical intervention( 1 Hz,5333 strain,4 h),L929 cells were divided into four groups: control group,experimental group with mechanical-trauma,PUN group,and mechanicaltrauma group of PUN pretreatment,and they were detected by flow cytometry to detect cell apoptosis rate. Western bolt was used to detect the protein expression levels of Bax、Bcl-2、caspase3、and Nrf2. And the changes of ROS level were detected by H2DCF-DA fluorescent probe. Results: The promotion of cell proliferation and the protein expression of Nrf2 were most significant at the concentration of 10 μmol/Land treating time of 24 h of PUN.Compared with the control group,apoptosis rate and ROS level of experimental group with mechanical-trauma were significantly increased. The pro-apoptosis protein Bax and Active-caspase3/caspase3 levels were significantly increased( P <0. 05). Compared with the experimental group with mechanical-trauma,cell apoptosis rate and ROS level decreased significantly,the anti-apoptosis protein Bcl2 increased significantly and the pro-apoptotic protein Bax and Active-caspase3/caspase3 levels decreased significantly( P <0. 05) in PUN pretreated group.Conclusions: PUN can significantly reduce apoptosis induced by mechanical-trauma,its mechanism may be related with the upregulation of the expression of antioxidant protein Nrf2. Thereby,the oxidative damage is inhibited by the increasing of ROS level induced by mechanical-trauma |
| Key words: Punicalagin Mechanical-trauma StressUrinaryIncontinence OxidativeDamage Apoptosis |
