文章摘要
陈 玮,殷 荔,缪爱梅,倪世宁,朱子阳,刘倩琦.促炎性细胞因子对大鼠胰岛β细胞miR-29家族及抗凋亡蛋白表达水平的影响[J].南京医科大学学报,2015,(6):787~792
促炎性细胞因子对大鼠胰岛β细胞miR-29家族及抗凋亡蛋白表达水平的影响
Effects of pro-inflammatory cytokines on expression levels of miR-29 family and anti-apoptotic proteins in rat pancreatic β cells
投稿时间:2015-01-12  
DOI:10.7655/NYDXBNS20150606
中文关键词: 促炎性细胞因子  miR-29  抗凋亡蛋白  凋亡  1型糖尿病
英文关键词: pro-inflammatory cytokines  miR-29  antiapoptotic proteins  apoptosis  type1 diabete
基金项目:南京市医药卫生科研课题(YKK12106)
作者单位
陈 玮 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
殷 荔 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
缪爱梅 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
倪世宁 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
朱子阳 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
刘倩琦 南京医科大学附属南京儿童医院内分泌科,江苏 南京 210008 
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中文摘要:
      目的:探讨促炎性细胞因子对胰岛β细胞miR-29家族及抗凋亡蛋白水平的影响。方法:将大鼠胰岛细胞系INS-1细胞在加或不加促炎性细胞因子混合物(IL-1β 10 ng/mL?TNF-α 50 ng/mL?IFN-γ 50 ng/mL)的培养液中培养24 h,设立对照组?促炎性细胞因子干预组。流式细胞仪测细胞凋亡,实时荧光定量PCR检测INS-1细胞miR-29a/b/c表达以及抗凋亡基因骨髓细胞白血病蛋白1(myeloid cell leukemia 1,Mcl-1)mRNA?B细胞淋巴瘤蛋白2(B-cell lymphoma 2,Bcl-2) mRNA的表达水平,Western blot技术检测Mcl-1?Bcl-2蛋白的表达情况。结果:①促炎性细胞因子干预的INS-1细胞miR-29a/b表达水平较正常对照组增加,差异有统计学意义(P < 0.05),miR-29c表达水平较正常对照组有上升趋势,但无统计学差异(P > 0.05);②促炎性细胞因子干预组INS-1细胞Mcl-1 mRNA?Bcl-2 mRNA表达水平较对照组减少,但差异无统计学意义(P > 0.05);③促炎性细胞因子干预组INS-1细胞抗凋亡蛋白Mcl-1?Bcl-2表达水平较正常对照组明显下降,差异有统计学意义(P < 0.01);④促炎性细胞因子干预组细胞凋亡率增高,与正常对照组相比差异有统计学意义(P < 0.05)。结论:促炎性细胞因子刺激处理的INS-1细胞miR-29a/b表达均上调,抗凋亡蛋白Mcl-1?Bcl-2下调,凋亡率上升,推测促炎性细胞因子可能通过调节miR-29家族及抗凋亡蛋白的表达水平诱导胰岛β细胞凋亡,从而促进1型糖尿病的发生。
英文摘要:
      Objective:To study the effects of pro-inflammatory cytokines on expression levels of microRNA-29 (miR-29) family and anti-apoptotic proteins in rat islet β cells. Methods: INS-1 cells, which belong to rat islet cells, were incubated in the presence or absence of a pro-inflammatory cytokine mixture (IL-1β10 ng/ml, TNF-α 50 ng/ml and IFN-γ50 ng/ml) for 24 h (the pro-inflammatory cytokine stimulated group and the control group). The function of INS-1 cells was evaluated by the glucose stimulating insulin releasing test. Cell apoptosis was detected by flow cytometry. The expression levels of miR-29 family and myeloid cell leukemia 1 (Mcl-1) mRNA, B-cell lymphoma (Bcl-2) mRNA in INS-1 cells were detected by real-time fluorescence quantitative PCR, and the protein expression levels of Mcl-1, Bcl-2 were detected by Western blot. Results:①The miR-29a and miR-29b expression levels were significantly increased in INS-1 cells stimulated by pro-inflammatory cytokine mixture,and the differences were statistically significant (P < 0.05). The expression level of miR-29c of the pro-inflammatory cytokine stimulated group was increased compared with the control group, but there was no statistical significance (P > 0.05). ②The mRNA expression levels of Mcl-1 and Bcl-2 in the pro-inflammatory cytokine stimulated group were decreased with no statistical significance (P > 0.05).③ In the pro-inflammatory cytokine stimulated group, the protein expression levels of Mcl-1 and Bcl-2 were downregulated, and the differences were statistically significant (P < 0.01). ④Pro-inflammatory cytokine mixture stimulated INS-1 cells led to a significant increase in cell death, and the difference was statistically significant (P < 0.05). Conclusion: Pro-inflammatory cytokines may downregulate the expressions of Mcl-1 and Bcl-2 through overexpressing miR-29 family, and then impact on apoptosis in pancreatic β cells, thereby trigger type 1 diabetes mellitus.
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