细胞因子对胰岛细胞凋亡及凋亡相关基因表达的影响  

Effects of Cytokines on Apoptosis and Expression of Apoptosis-Related Genes in Rat Islet Cells

作　　者：何庆[1] 刘铭[2] 于德民[3] 苏京[4] 王保平[4] 王晶[4] 陈克勤[4] 尹潍[1] 

机构地区：[1]天津医科大学总医院内分泌科,300052 [2]天津医科大学总医院老年病研究室,300052 [3]天津医科大学代谢病医院 [4]丹麦诺和诺德中国研究发展中心

出　　处：《天津医药》2009年第1期39-42,共4页Tianjin Medical Journal

基　　金：天津市自然科学基金资助课题(项目编号:953608411;983702411;043607911)

摘　　要：目的:观察肿瘤坏死因子α(TNFα)和干扰素γ(IFNγ)对大鼠胰岛细胞和小鼠βTC-3细胞凋亡和凋亡相关基因表达的影响。方法:应用TUNEL法检测高浓度TNFα和IFNγ培养后大鼠胰岛细胞和小鼠βTC-3细胞凋亡百分率,应用半定量RT-PCR(sQRT-PCR)检测培养后bcl-2、bax、c-myc、p53和fas mRNA的表达。结果:大鼠胰岛细胞经高浓度TNFα和IFNγ处理后,上清液中胰岛素浓度明显下降,由(169.69±1.62)mIU/L降为(136.1±1.44)mIU/L(P<0.01);凋亡细胞比例明显增加,由(10.25±1.71)%升为(17.25±1.71)%(P<0.01)。βTC-3细胞经高浓度TNFα和IFNγ处理后,上清液中胰岛素浓度明显下降,由(254.13±4.35)mIU/L降为(221.42±3.98)mIU/L(P<0.01);凋亡细胞比例明显增加,由(20.25±2.22)%升为(38.75±3.4)%(P<0.01)。大鼠胰岛细胞和βTC-3细胞经高浓度TNFα和IFNγ处理后,致凋亡基因bax、c-myc、fas mRNA表达明显增加(P<0.05);抗凋亡基因bcl-2 mRNA表达明显下降(P<0.05);bcl-2/bax mRNA表达比率明显下降(P<0.01);p53mRNA表达无明显改变(P>0.05)。结论:TNFα和IFNγ可能通过诱导胰岛细胞凋亡导致或加重糖尿病,其中bcl-2/bax mRNA表达比率变化可能起重要作用。Objective：To investigate effects of tumor necrosis factor （TNF） α and interferon （IFN）γ on the apoptosis and expression of apoptosis-related genes in islet cells. Methods：Apoptotic cells of primary rat islet cell and β TC-3 cell line treated by high level of TNFα and IFNγ were identified by TUNEL method. The mRNA levels of bcl-2, bax, c-myc, p53 and fas were evaluated by the method of semi-quantitative RT-PCR. Results： High levels of TNFet and IFNγdecreased the insulin level in the supernatant of primary rat islet cells [ （ 169.69± 1.62） mIU/L to （ 136.1 ± 1.44） mIU/L, P 〈 0.01 ]and βTC-3 cell line [（254.13±4.35） mlU/L to （221.42±3.98） mIU/L, P 〈 0.01]. And also the same treatment increased the proportion of the apoptotic cell of primary rat islet cell[（ 10.25±1.71 ）% to （ 17.25±1.71 ）% ,P 〈 0.01] and βTC-3 cell line [（20.25±2.22）% to （38.75±3.4）%, P〈 0.01]. After the treatment of high level of TNFα and IFNγ the mRNA expression of the pro-apoptosis genes, such as bax, c-myc and fas, were increased significantly （P 〈 0.05）, while anti-apoptosis gene bel-2 mRNA＇s expression was decreased signifieantly（P 〈 0.05 ）. The mRNA expression ratio between bcl-2 and bax decreased significantly （P 〈 0.01）. The expression of p53＇s mRNA was not changed significantly （P 〉 0.05）. Conclusion： High levels of TNF and IFN± may initiate and exacerbate the diabetes through inducing the apoptosis of islet cells. The change of bel-2/bax mRNA expression ratio may play an important role in the apoptosis.

关 键 词：胰岛 细胞凋亡 肿瘤坏死因子Α 干扰素Γ 基因表达 大鼠 Sprague—Dawley 

分 类 号：R285.5[医药卫生—中药学] R587.1[医药卫生—中医学]