雌激素对SD大鼠肾脏缺血再灌注肾小管上皮细胞Cx43蛋白表达的影响

doi:10.19586/j.2095-2341.2022.0023

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 452-459.DOI: 10.19586/j.2095-2341.2022.0023

雌激素对SD大鼠肾脏缺血再灌注肾小管上皮细胞Cx43蛋白表达的影响

马义博¹(), 常越辰¹, 马克涛¹, 李丽¹, 王勤章², 李应龙²^,³()

^1.石河子大学医学院，新疆石河子 832000
^2.石河子大学医学院第一附属医院泌尿外科，新疆石河子 832000
^3.成都中医药大学附属第五人民医院泌尿外科，成都 611130

收稿日期:2022-03-03 接受日期:2022-03-15 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 李应龙
作者简介:马义博 E-mail：674329609@qq.com；
基金资助:
国家自然科学基金项目(81960188);石河子大学自主资助支持项目(ZZZC202025A)

Effects of Estrogen on the Expression of Cx43 Protein in Renal Tubular Epithelial Cells of SD Rats with Ischemia⁃reperfusion

Yibo MA¹(), Yuechen CHANG¹, Ketao MA¹, Li LI¹, Qinzhang WANG², Yinglong LI²^,³()

^1.Medical College，Shihezi University，Xinjiang Shihezi 832000，China
^2.Department of Urology，the First Affiliated Hospital of Medical College，Shihezi University，Xinjiang Shihezi 832000，China
^3.Department of Urology，the Fifth People's Hospital Affiliated to Chengdu University of Traditional Chinese Medicine，Chengdu 611130，China

Received:2022-03-03 Accepted:2022-03-15 Online:2022-05-25 Published:2022-05-26
Contact: Yinglong LI

摘要/Abstract

摘要：

为了探讨雌激素（estrogen，E₂）对SD大鼠肾脏缺血再灌注（ischemia-reperfusion，I/R）肾小管上皮细胞Cx43蛋白表达的影响，选取32只健康雌性SD大鼠作为实验材料，所有雌性SD大鼠均先实施去卵巢（ovariectomized，OVX）处理，之后32只雌性OVX SD大鼠随机分为OVX组、OVX+假手术组、OVX+I/R组、OVX+I/R+E₂组，每组8只。去卵巢2周后，OVX组不进行任何处理，OVX+Sham组进行假手术处理，OVX+I/R组进行缺血再灌注处理，OVX+I/R+E₂组在进行缺血再灌注处理前，连续3 d予以雌激素处理。通过比较各组SD大鼠血肌酐水平、尿素氮水平、肾脏HE染色及Paller评分，评价肾脏组织损伤程度，并通过免疫组化和蛋白印迹法分析各组SD大鼠肾小管上皮细胞中Cx43蛋白的表达位置及水平。与OVX组比较，OVX+I/R组血清BUN、SCr水平明显升高（P<0.05）；与OVX+I/R组比较，OVX+I/R+E₂组血清BUN、SCr水平明显降低（P<0.05）。HE染色观察发现，与OVX组相比，OVX+I/R组肾小管扩张明显，部分细胞碎裂坏死，Paller评分明显升高（P<0.05）；与OVX+I/R组相比，OVX+I/R+E2组可见肾小管轻度扩张，细胞结构较为完整，Paller评分明显下降（P<0.05）。通过免疫组化发现，OVX+I/R组Cx43蛋白在肾小管上皮细胞中阳性表达高于OVX组及OVX+Sham组（P<0.05），OVX+I/R+E₂组Cx43蛋白表达低于OVX+I/R组（P<0.05）。Western blotting结果显示，与OVX组相比，OVX+I/R组Cx43蛋白在肾小管上皮细胞中表达量增加，肾脏损伤后肾小管上皮细胞Cx43表达明显上调（P<0.05）；与OVX+I/R组相比，OVX+I/R+E₂组经过雌激素干预后，肾脏功能损伤减轻，肾小管上皮细胞Cx43的蛋白表达量下调（P<0.05）。结果表明雌激素可减轻肾缺血再灌注损伤造成的肾脏损伤，其可能是通过调节肾小管上皮细胞中Cx43的表达实现的。研究结果为临床治疗肾脏缺血再灌注损伤提供了新的思路。

关键词: 雌激素, SD大鼠, 肾脏缺血再灌注损伤, 肾小管上皮细胞, Cx43蛋白

Abstract:

To explore the effect of estrogen （estrogen， E₂） on the expression of Cx43 protein in renal tubular epithelial cells of SD rats with ischemia-reperfusion （I/R）， 32 healthy female SD rats were selected as experimental materials. All female SD rats were first subjected to ovariectomized （OVX） treatment， the 32 female OVX SD rats were randomly divided into OVX group， OVX+sham operation group， OVX+ I/R group， OVX+I/R+E₂ group， with 8 animals in each group. After 2 weeks of ovariectomy， the OVX group received no treatment， the OVX+Sham group received sham operation， the OVX+I/R group received ischemia-reperfusion treatment， and the OVX+I/R+ E₂ group was treated with estrogen for 3 days before ischemia-reperfusion treatment. By comparing the levels of serum creatinine， blood urea nitrogen， renal HE staining and Paller score of SD rats in each group， the degree of renal tissue damage was evaluated， and the expression position and level of Cx43 protein in renal tubular epithelial cells of SD rats in each group were analyzed by immunohistochemistry and Western blotting. Compared with the OVX group， the serum BUN and SCr levels in the OVX+I/R group were significantly increased （P<0.05）. Compared with the OVX+I/R group， the serum BUN and SCr levels in the OVX+I/R+E₂ group were significantly decreased （P<0.05）. HE staining showed that， compared with the OVX group， the renal tubules in the OVX+I/R group were significantly enlarged， some cells were fragmented and necrotic， and the Paller score was increased （P<0.05）. Compared with the OVX+I/R group， the renal tubules in the OVX+I/R+E₂ group were slightly expanded， the cell structure was more complete， and the Paller score was decreased （P<0.05）. Compared with OVX group and OVX+Sham group， the positive expression in renal tubular epithelial cells was significantly increased （P<0.05）， and the expression of Cx43 protein in OVX+I/R+E₂ group was lower than that in OVX+I/R group （P<0.05）. The results of Western blotting showed that， compared with the OVX group， the expression of Cx43 protein in the renal tubular epithelial cells in the OVX+I/R group was increased （P<0.05）， and the expression of Cx43 in the renal tubular epithelial cells was significantly up-regulated after kidney injury （P<0.05）. Compared with OVX+I/R group， after estrogen intervention in OVX+I/R+E₂ group， renal function damage was alleviated， and the protein expression of Cx43 in renal tubular epithelial cells was down-regulated （P<0.05）. The results of this study showed that， estrogen could alleviate renal injury caused by renal ischemia-reperfusion injury， which might be achieved by regulating the expression of Cx43 in renal tubular epithelial cells. The result of this paper provided new ideas for clinical treatment of renal ischemia-reperfusion injury.

Key words: estrogen, SD rat, renal ischemia-reperfusion injury, renal tubular epithelial cells, Cx43 protein

中图分类号:

R692

马义博, 常越辰, 马克涛, 李丽, 王勤章, 李应龙. 雌激素对SD大鼠肾脏缺血再灌注肾小管上皮细胞Cx43蛋白表达的影响[J]. 生物技术进展, 2022, 12(3): 452-459.

Yibo MA, Yuechen CHANG, Ketao MA, Li LI, Qinzhang WANG, Yinglong LI. Effects of Estrogen on the Expression of Cx43 Protein in Renal Tubular Epithelial Cells of SD Rats with Ischemia⁃reperfusion[J]. Current Biotechnology, 2022, 12(3): 452-459.

图/表 3

图1 各组大鼠BUN和SCr水平比较注：*表示与OVX相比，差异在P<0.05水平上有统计学意义；#表示OVX+I/R+E2组与OVX+I/R组相比，差异在P<0.05水平上有统计学意义。

Fig.1 Comparison of the levels of BUN and SCr among different groups of rats

图2 各组大鼠肾组织病理改变和Paller评分比较A：HE染色结果。光镜放大倍数为200倍。B：Paller评分。*表示与OVX组相比，差异在P<0.05水平上有统计学意义；#表示OVX+I/R+E2组与OVX+I/R组相比，差异在P<0.05水平上有统计学意义。

Fig.2 Comparison of histopathological changes in kidney tissues and Paller scores among different groups of rats

图3 Cx43在各组大鼠肾小管上皮细胞中的表达A：免疫组化结果；B：蛋白印迹检测结果；C：Cx43蛋白表达水平。*表示与OVX相比，差异在P<0.05水平上有统计学意义；#表示OVX+I/R+E2组与OVX+I/R组相比，差异在P<0.05水平上有统计学意义。

Fig.3 The expression of Cx43 in renal tubular epithelial cells among different groups of rats

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雌激素对SD大鼠肾脏缺血再灌注肾小管上皮细胞Cx43蛋白表达的影响

Effects of Estrogen on the Expression of Cx43 Protein in Renal Tubular Epithelial Cells of SD Rats with Ischemia⁃reperfusion

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