Effect of mitofusin 2 overexpression on the proliferation and apoptosis of high-glucose-induced rat glomerular mesangial cells
Effect of mitofusin 2 overexpression on the proliferation and apoptosis of high-glucose-induced rat glomerular mesangial cells
复制标题
DOI：
10.5301/jn.5000089
复制
发表时间：
2012-11-01
影响因子：
3.4
通讯作者：
Fu Ping
中科院分区：
文献类型：
--
作者：
Tang Wan-Xin;Cui Tian-Lei;Fu Ping
背景：Mitofusin 2 （Mfn2）调节线粒体形态和相关信号通路。然而，Mfn2在肾脏疾病中的作用尚不完全清楚。本研究旨在探讨Mfn2过表达对高糖诱导的细胞增殖和凋亡的影响。另一个目的是探索大鼠肾小球系膜细胞（GMC）系可能潜在的信号转导机制。方法：转染腺病毒介导的Mfn2基因后，采用实时荧光定量PCR检测Mfn2的表达。研究高糖诱导Mfn2及相关蛋白浓度随时间的变化，确定最佳研究时间。Western blot检测Mfn2、增殖细胞核抗原（PCNA）、p-Akt、p-ERK1/2、Bcl-2蛋白表达水平。CCK-8法检测细胞增殖。流式细胞术检测细胞凋亡。结果：腺病毒介导Mfn2基因成功表达。高糖刺激48小时后，Mfn2表达与p-ERK1/2和磷酸化Akt的相关性显著。高糖升高p-ERK1/2，而过表达Mfn2抑制p-ERK1/2。PCNA和GMC增殖水平的变化与p-ERK1/2一致。过表达Mfn2显著抑制Akt磷酸化和Bcl-2的表达。AdMfn2组细胞凋亡率升高。结论：过表达Mfn2可减轻GMC细胞增殖，增加细胞凋亡。随后，细胞数量得以维持，这可能有助于逆转早期糖尿病肾病的病理改变。Mfn2可能通过MAPK/ERK和PI3K/Akt信号通路发挥作用，与细胞增殖和凋亡相关。
Background: Mitofusin 2 (Mfn2) regulates mitochondrial morphology and associated signaling pathways. However, the role of Mfn2 in kidney disease is not fully understood. The present study aimed to investigate the effects of Mfn2 overexpression on high-glucose-induced cell proliferation and apoptosis. Another objective was to explore the possible underlying signal transduction mechanisms in a rat glomerular mesangial cell (GMC) line.Methods: After adenovirus-mediated Mfn2 gene transfection, Mfn2 expression was detected by real-time PCR. Time-dependent concentration changes in Mfn2 and relevant proteins induced by high glucose were investigated to define the optimal time for research. The protein expression levels of Mfn2, proliferating cell nuclear antigen (PCNA), p-Akt, p-ERK1/2 and Bcl-2 were examined on Western blots. Cell proliferation was detected by the CCK-8 method. Apoptosis was analyzed by flow cytometry.Results: Mfn2 gene expression was successfully increased via adenovirus mediation. The correlation of Mfn2 expression with p-ERK1/2 and phosphorylated Akt was significant 48 hours after high-glucose stimulation. p-ERK1/2 was increased by high glucose, but was inhibited by overexpressed Mfn2. Changes in the PCNA and GMC proliferative level coincided with p-ERK1/2. Overexpressed Mfn2 significantly inhibited Akt phosphorylation and Bcl-2 expression. The apoptosis rate increased in the AdMfn2 group.Conclusions: Overexpressed Mfn2 could alleviate GMC proliferation and increase apoptosis. Subsequently, cellular quantity is maintained, which may contribute to reversing early diabetic nephropathy pathological changes. Mfn2 may perform its activities through the MAPK/ERK and PI3K/Akt signal pathways in correlation with proliferation and apoptosis.