虎杖苷对四氧嘧啶诱导的大鼠胰岛细胞损伤的保护作用

Protective Effect of Polydatin on Alloxan-induced INS-1 Cell Damage

目的:研究虎杖苷(Polydatin)对四氧嘧啶(Alloxan)诱导的大鼠胰岛素瘤INS-1细胞损伤的保护作用。方法:利用细胞增殖实验分别检测四氧嘧啶或虎杖苷对INS-1细胞增殖的影响,确定给药浓度;实验分为空白组、模型组(18 mmol/L四氧嘧啶)和不同浓度虎杖苷保护组(分别给予25、50μg/mL虎杖苷,同时给予18 mmol/L四氧嘧啶),检测细胞存活率、LDH释放、胰岛素分泌、凋亡相关Caspase-3和Caspase-9的活性、活性氧簇ROS、一氧化氮NO、丙二醛MDA的含量及抗氧化相关超氧化物歧化酶SOD、还原型谷胱甘肽GSH水平。结果:10~30 mmol/L四氧嘧啶处理使INS-1细胞存活率显著降低(P<0.05),18 mmol/L四氧嘧啶处理细胞贴壁数目减少,LDH释放升高,胰岛素分泌量降低,凋亡相关Caspase-3和Caspase-9的活性显著升高(P<0.05),氧化应激相关ROS、NO和MDA含量显著提高(P<0.05),抗氧化相关SOD和GSH水平显著降低(P<0.05)。5~100μg/mL虎杖苷对INS-1细胞无毒性作用,相较于模型组,25和50μg/mL虎杖苷能显著提高四氧嘧啶诱导INS-1细胞存活率,改善细胞形态,降低LDH活性,增加胰岛素分泌量,抑制凋亡相关Caspase-3和Caspase-9酶活性,抑制四氧嘧啶诱导的氧化损伤,提高抗氧化相关酶活力(P<0.05)。结论:虎杖苷对四氧嘧啶诱导的INS-1细胞损伤有显著保护作用,其作用机制可能与抑制氧化应激及凋亡有关。

Objective:To investigate the protective effect of polydatin on damage of INS-1 cells induced by alloxan. Methods:MTT assay was used to detect the survival rate of INS-1 cells treated with alloxan or polydatin respectively to select the concentration of alloxan or polydatin used for further experiments. INS-1 cells were divided into control group,model group(18 mmol/L alloxan),and polydatin groups(25 or 50 μg/mL+18 mmol/L alloxan). Cell survival rates were measured by MTT method;LDH release was detected;insulin secretion was tested;the activities of caspase-3 and caspase-9 were analyzed;supernatant NO,intracellular ROS,MDA level,SOD activities,GSH contents were measured by assay kits. Results:10~30 mmol/L alloxan significantly decreased the cell survival rate in contrast to the control group(P<0.05);the polydatin(25,50 μg/mL)significantly improved INS-1 cells survival rates in contrast to control group(P<0.05). Compared with model group,polydatin(25,50 μg/mL)significantly increased INS-1 cells survival rates,improved the cell morphology;reduced the release of LDH,increased insulin secretion;inhibited both caspase-3 and caspase-9 activities(P<0.05);polydatin also suppressed the production of ROS,NO and MDA level,significantly elevated SOD activity and GSH in alloxan-damaged INS-1 cells(P<0.05). Conclusion:Polydatin can effectively improve the damage of INS-1 cells induced by alloxan and its antioxidative and anti-apoptotic effect may serve as possible mechanism.