黄芪甲苷抑制高糖腹透液诱导HMrSV5氧化应激与EMT的实验研究

史俊1; 俞曼殊1; 杨劲松1; 盛梅笑2; 3*

文章摘要

史俊1，俞曼殊1，杨劲松1，盛梅笑2，3*.黄芪甲苷抑制高糖腹透液诱导HMrSV5氧化应激与EMT的实验研究[J].南京中医药大学学报(社会科学版),2016,32(4):337-341.

黄芪甲苷抑制高糖腹透液诱导HMrSV5氧化应激与EMT的实验研究

The Effect of Astragaloside-Ⅳ on Oxygen Stress and EMT of HMrSV5 Induced by High-glucose Peritoneal Dialysate

DOI：

中文关键词: 黄芪甲苷高糖腹透液腹膜纤维化 PMCs EMT ROS

英文关键词: Astragaloside-Ⅳ (AS-Ⅳ) peritoneal dialysate peritoneal fibrosis EMT ROS

基金项目:

作者	单位
史俊1，俞曼殊1，杨劲松1，盛梅笑2，3*	1.南京中医药大学第一临床医学院，江苏南京 210023；2.南京中医药大学附属医院，江苏南京 210029；3.南京中医药大学肾病研究所，江苏南京 210029

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中文摘要:

目的观察黄芪甲苷（Astragaloside Ⅳ，AS-Ⅳ）对高糖腹透液诱导人腹膜间皮细胞HMrSV5氧化应激及EMT的影响，探讨ROS在EMT中的作用及AS-Ⅳ干预机制。方法采用葡萄糖腹透液诱导建立HMrSV5氧化应激模型，相差显微镜观察细胞形态变化，MTT检测AS-Ⅳ及葡萄糖腹透液对细胞活力的影响；予40μg/mL AS-Ⅳ干预HMrSV5氧化应激，DHE荧光探针检测药物作用前后ROS生成水平变化，Western blot检测EMT相关蛋白变化；与ROS清除剂NAC作对照，观察AS-Ⅳ干预HMrSV5氧化应激EMT指标及Smads蛋白的变化。结果 ①40、50μg/mL AS-Ⅳ干预后细胞活力稍有上升（P<0.05）；随腹透液作用时间的延长及葡萄糖浓度提高，细胞活力明显抑制，以4.25%高糖腹透液干预48h最明显（P<0.05），细胞拉伸变长；②40μg/mL AS-Ⅳ作用细胞能抑制形态改变，降低模型组ROS的生成（P<0.05），上调E-cadherin、ZO-1表达，下调α-SMA表达，并抑制Smad2/3磷酸化；③5mmol/L NAC产生类似调控作用。结论高糖腹透液可抑制PMCs细胞活性，增加ROS生成，导致PMCs EMT，其分子机制可能与ROS参与调控的Smads通路激活有关，而AS-Ⅳ可能通过减少ROS生成，抑制Smad2/3磷酸化，阻抑PMCs EMT。

英文摘要:

OBJECTIVE To observe the effect of Astragaloside-Ⅳ (AS-Ⅳ) on oxygen stress and EMT of peritoneal mesothelial cells (PMCs) HMrSV5， explore the function of ROS in EMT and regulatory mechanism of AS-Ⅳ. METHODS We used peritoneal dialysis solutions (PDS) with high glucose concentration to establish oxygen stress cell model of HMrSV5. Morphologic alterations of cells were observed and MTT assay was used to detect cell viability. DHE fluorescent probes were loaded to label secreted ROS. EMT-associated proteins were expressed by western blotting analysis. RESULTS ①40 or 50μg/mL AS-Ⅳ increased the viability of HMrSV5 (P<0.05). Cell viability was supressed due to the longstanding effect of PDS and high glucose concentration， notably in intervention of PDS containing 4.25% glucose for 48h. Phenotypic changes of HMrSV5 characterized by cell elongation and losing cobblestone like feature. ②40μg/mL AS-Ⅳ could suppress morphologic changes of cells， decrease ROS expression in model group (P<0.05)， upregulate the protein expression of E-cadherin and ZO-1 while downregulate α-SMA. Furthermore， the phosphorylation of Smad2/3 protein was inhibited. ③ 5mmol/L NAC had the similar effect on EMT of PMCs. CONCLUSION PDS with high glucose concentration can suppress cell viability and induce EMT of PMCs by upregulating ROS expression and activation of Smads pathway. AS-Ⅳ can reduce ROS generation， suppress the phosphorylation of Smad2/3 and thus， inhibite the EMT of PMCs.

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