急性肾损伤miR-34a、GPS2表达变化及其对JNK通路的影响

doi:10.3969/j.issn.2095-9400.2020.06.002

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摘要目的：探讨急性肾损伤miR-34a、GPS2表达变化及其对相关通路指标p-JNK的影响。方法：30只SD大鼠随机分为灌注对照组（0 h、24 h、48 h）和灌注模型组（0 h、24 h、48 h）（每组n=5），分别收集各组大鼠的血清和肾脏组织。将HEK 293T细胞分为对照组（Control、NC inhibitor）和模型组（Model、Model+miR-34a inhbitor），收集细胞提取RNA和蛋白。采用ELISA试剂盒测定各组造模前后血清中TNF-α和TGF-β水平，HE染色比较肾脏组织损伤病理变化，RT-qPCR法检测各组肾脏组织和细胞中miR-34a、GPS2的mRNA表达变化，Western blot检测肾脏组织和细胞中GPS2、p-JNK的蛋白表达变化。结果：HE染色结果显示，与各自对照组相比，模型组肾小管出现损伤改变，包括上皮细胞肿胀、刷状缘消失、空泡形成。ELISA结果显示，随着灌注时间的增加，模型组TNF-α和TGF-β水平逐步增高（P＜0.01），模型组各时段TNF-α和TGF-β水平均明显高于对照组（P＜0.01）。大鼠肾组织PCR和Western blot结果显示，与对照组相比，模型组同时段miR-34a、p-JNK表达量均升高（P＜0.05），而GPS2明显下降（P＜0.05）。细胞PCR和Western blot结果显示，与对照组相比，沉默miR-34a基因后，GPS2明显升高（P＜0.05），而p-JNK表达量下降（P＜0.05）。结论：miR-34a可能通过GPS2间接激活JNK通路，促进并加重急性肾损伤。

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	陈伟伟
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关键词 ：肾脏缺血, 再灌注损伤, 肾小管炎症因子, G蛋白途径抑制因子2, JNK信号通路

Abstract：Objective: To explore the expression changes of miR-34a and GPS2 and their effects on p-JNK in ischemia-reperfusion and hypoxia/reoxygenation models. Methods: Thirty SD rats were randomly divided into 6 groups (n=5). Serum and kidney tissues were collected from the perfusion control group (0 h, 24 h, 48 h) and the perfusion model group (0 h, 24 h, 48 h). HEK 293T cells were divided into control group (Control, NC inhibitor) and model group (Model, Model+miR-34a inhbitor). RNA and protein were extracted from the cells.The levels of tumor necrosis factor (TNF-α) and transforming growth factor (TGF-β) in serum were measured by ELISA kit. The pathological changes of kidney tissues were compared by hematoxylin-eosin (HE). The mRNA changes of miR-34a and GPS2 were detected by RT-qPCR, and the protein expressions of GPS2 and p-JNK were detected by Western blot. Results: HE results showed that renal tubular injury in the model group had epithelial cell swelling, brush edge disappearance and vacuolation formation, compared with the control group. ELISA results showed that there was no difference in TNF-α and TGF-β levels between the model group and the control group before and after the establishment of the model. TNF-α and TGF-β levels in the model group increased with the perfusion time and significantly higher than the respective control group (P<0.01). Compared with the control group, the damage of renal tubules in the model group had swelling of epithelial cells, disappearance of brush border and formation of vacuoles. The expressions of miR-34a and p-JNK increased in the model group (P<0.05), while GPS2 decreased (P<0.05). Cell PCR and Western blot show that GPS2 increased significantly (P<0.05) and p-JNK decreased significantly (P<0.05) after miR-34a was silenced. Conclusion: miR-34a may indirectly activate JNK pathway through GPS2, and then promotes and aggravates the occurrence of acute kidney injury.

Key words： renal ischemia reperfusion injury renal tubular inflammatory factor G protein pathway supppressor 2 JNK pathway

收稿日期: 2019-09-15

基金资助:浙江省自然科学基金资助项目（LY17H050006）；浙江省医药卫生科技计划项目（2017KY107）；国家自然科学基金青年基金资助项目（81200513）。

通讯作者: 蔡晖，教授，主任医师，Email：hhcai62@gmail.com。

作者简介: 陈伟伟（1993-），女，浙江温州人，硕士生。

链接本文:

https://xb.wmu.edu.cn/CN/10.3969/j.issn.2095-9400.2020.06.002 或 https://xb.wmu.edu.cn/CN/Y2020/V50/I6/438

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