肾小管上皮细胞G2-M期阻滞在缺氧诱导的肾间质纤维化中的作用

发布时间：2018-05-05 09:23

本文选题：缺氧 + G-M阻滞　；参考：《东南大学学报(医学版)》2017年02期

【摘要】：目的:探讨肾小管上皮细胞G2-M期阻滞在缺氧诱导的肾间质纤维化中的作用。方法:体外实验,人肾小管上皮细胞(HK2)分别置于常氧21%和缺氧1%的孵箱里,培养24,48 hrs。采用流式细胞术检测缺氧48 hrs后上皮细胞的周期分布;采用Western blot法检测Collagen4A1(COL4A1)及α-平滑肌肌动蛋白(α-SMA)的蛋白表达水平。单侧输尿管梗阻(UUO)14 d小鼠模型,HE及Masson染色观察肾组织病理改变及纤维化程度,免疫组化染色观察α-SMA的表达及部位;Western blot检测低氧标记分子HIF-1α、细胞周期蛋白cyclinB1和cyclinD1的蛋白水平。结果:与常氧组比较,缺氧组肾小管上皮细胞G2/M期比例增高(P0.05),同时α-SMA和COL4A1的蛋白表达水平增高(P0.05);与假手术组比较,HE及Masson染色显示模型组肾组织纤维化程度增高,α-SMA的表达增高且主要分布在间质中;Western blot结果显示:HIF-1α及G2/M期标记物cyclinB1/cyclinD1比值增加。结论:肾小管上皮细胞G2-M期阻滞参与缺氧诱导的肾间质纤维化。
[Abstract]:Aim: to investigate the role of G 2-M arrest in renal interstitial fibrosis induced by hypoxia. Methods: in vitro, human renal tubular epithelial cells (HK2) were incubated in a incubator containing 21% normoxic and 1% hypoxia, respectively. Flow cytometry was used to detect the distribution of epithelial cell cycle after 48 hrs hypoxia, and Western blot assay was used to detect the protein expression of Collagen4A1COL4A1 and 伪 -smooth muscle actin (伪 -SMAs). The pathological changes of renal tissue and the degree of renal fibrosis were observed by HE and Masson staining. The expression of 伪 -SMA and the protein levels of HIF-1 伪, cyclin cyclinB1 and cyclinD1 were detected by immunohistochemical staining. Results: compared with normoxic group, Compared with sham-operated group, the ratio of G _ 2 / M phase of renal tubular epithelial cells in hypoxia group and the protein expression level of 伪 -SMA and COL4A1 in renal tubular epithelial cells were increased and the expression of 伪 -SMA and 伪 -SMA in model group was higher than that in sham operation group, and the expression of 伪 -SMA and 伪 -SMA in model group was higher than that in sham-operated group. The results of Western blot in the interstitial tissue showed that the ratio of cyclinB1/cyclinD1 in the G 2 / M phase marker and the ratio of 1% HIF-1 伪 and G 2 / M phase marker were increased. Conclusion: G 2-M phase block of renal tubular epithelial cells is involved in hypoxia-induced renal interstitial fibrosis.
【作者单位】：第四军医大学西京医院肾内科国家肿瘤重点实验室;军事口腔医学国家重点实验室口腔疾病国家临床医学研究中心陕西省口腔生物工程技术研究中心第四军医大学口腔医学院麻醉科;第四军医大学学员一旅;海军总医院肿瘤诊疗中心;
【基金】：国家自然科学基金面上项目(NO.81370789)
【分类号】：R692

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