NIPA2通过JAK/STAT信号通路对高糖诱导的成骨细胞凋亡的调控作用及其机制

doi:10.13481/j.1671-587x.20200212

摘要/Abstract

摘要： 目的：探讨普瑞德-威利/安格曼综合征区域蛋白2（NIPA2）对高糖诱导的成骨细胞凋亡的影响，阐明其作用机制。方法：将成骨细胞MC3T3-E1采用26 nmol·L^-1高糖处理24 h，将细胞分为HG+si-con组（转染si-con）、HG+si-NIPA2组（转染si-NIPA2）、HG+si-NIPA2+DMSO组（转染si-NIPA2后再用DMSO处理）和HG+si-NIPA2+AG490组（转染si-NIPA2后再用AG490处理），另设对照组。采用脂质体法转染MC3T3-E1细胞，再用26 nmol·L^-1高糖处理；采用qRT-PCR法检测MC3T3-E1细胞中NIPA2 mRNA表达水平，Western blotting法检测细胞中NIPA2、P21、Cleavedcaspase-3、p-JAK2和p-STAT3蛋白表达水平，MTT法检测细胞增殖活性，流式细胞术检测细胞凋亡率。结果：与对照组比较，HG组MC3T3-E1细胞中NIPA2 mRNA和蛋白表达水平均明显降低（P<0.01）。敲减NIPA2后，与HG+si-con组比较，HG+si-NIPA2组MC3T3-E1细胞中NIPA2蛋白表达水平明显降低（P<0.01），P21和Cleaved caspase-3蛋白表达水平明显升高（P<0.01），MC3T3-E1细胞在48和72 h时细胞增殖活性明显降低（P<0.01），细胞凋亡率明显升高（P<0.01），JAK/STAT信号通路相关蛋白p-JAK2和p-STAT3的表达水平明显升高（P<0.01）。与HG+si-NIPA2+DMSO组比较，HG+si-NIPA2+AG490组MC3T3-E1细胞中NIPA2蛋白表达水平明显升高（P<0.01），P21和Cleaved caspase-3蛋白表达水平明显降低（P<0.01），在48和72 h时细胞增殖活性明显升高（P<0.01），细胞凋亡率明显降低（P<0.01）。结论：NIPA2对高糖诱导的成骨细胞增殖和凋亡具有调控作用，其调控机制与JAK/STAT信号通路有关。

关键词: NIPA2, JAK/STAT信号通路, 成骨细胞, 细胞凋亡

Abstract: Objective: To investigate the effect of Prader-Willi/Angelman syndrome region protein 2(NIPA2) on the high glucose-induced osteoblast apoptosis, and to elucidate its mechanism; Methods: The osteoblasts MC3T3-E1 were treated with 26 nmol·L^-1 high glucose for 24 h; the cells were divided into HG+si-con group (transfected with si-con), HG+si-NIPA2 group (transfected with si-NIPA2), HG+si-NIPA2+DMSO group (transfected with si-NIPA2 and treated with DMSO), HG+si-NIPA2+AG490 (transfected with si-NIPA2 and treated with AG490),at the same time control group was set up. After the MC3T3-E1 cells were transfected by liposome method, they were treated with 26 nmol·L^-1 high glucose again.The qRT-PCR method was used to detect the expression level of NIPA2 mRNA in theMC3T3-E1 cells;Western blotting method was used to determine the expression levels of NIPA2, P21, Cleaved caspase-3, p-JAK2 and p-STAT3 proteins in the MC3T3-E1 cells; MTT method was used to detect the cell proliferation activities, and flow cytometry was used to measure the apoptotic rates. Results: Compared with control group, the mRNA and protein expression levels of NIPA2 in the MC3T3-E1 cells in HG group were significantly decreased (P<0.01). After knocking down the NIPA2, compared with HG+si-con group, the expression level of NIPA2 protein in HG+si-NIPA2 group was significantly decreased (P<0.01), the expression levels of P21 and Cleaved caspase-3 proteins were significantly increased (P<0.01), the proliferation activities of MC3T3-E1 cells were significantly decreased at 48 and 72 h (P<0.01), the apoptotic rate was significantly increased (P<0.01), and the expression levels of JAK/STAT signaling pathway-related proteins p-JAK2 and p-STAT3 were significantly increased (P<0.01).Compared with HG+si-NIPA2+DMSO group, the expression levels of NIPA2 protein in HG+si-NIPA2+AG490 group was significantly increased (P<0.01), the expression levels of P21 and Cleaved caspase-3 proteins were significantly decreased (P<0.01), the cell proliferation activities were significantly increased at 48 and 72 h (P<0.01), and the apoptotic rate was significantly decreased (P<0.01). Conclusion: NIPA2 can regulate the proliferation and apoptosis of the osteoblasts induced by high glucose, and its regulatory mechanism is related to the JAK/STAT signaling pathway.

Key words: NIPA2, JAK/STAT signaling pathway, osteoblasts, apoptosis

中图分类号:

R329.2

吴玉洁, 邢学农. NIPA2通过JAK/STAT信号通路对高糖诱导的成骨细胞凋亡的调控作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(02): 280-285.

WU Yujie, XING Xuenong. Regulatory effect of NIPA2 on high glucose-induced osteoblast apoptosis via JAK/STAT signaling pathway and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 280-285.

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