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目的探讨高糖刺激下转染过表达人胰岛淀粉样多肽基因的大鼠胰岛素瘤细胞(INS-1)细胞凋亡的影响及其相关基因表达。方法应用酶联免疫吸附法(ELISA)检测高糖刺激后INS-1及转染人胰岛淀粉样多肽的INS-1细胞(h IAPP/INS-1)上清液的胰岛素水平;利用Hoechst33258染色检测细胞凋亡百分率;采用RTPCR方法检测高糖刺激培养后的Bcl2、Bax m RNA的水平。结果高糖刺激下,与未转染人胰岛淀粉样多肽INS-1细胞相比较,h IAPP/INS-1细胞胰岛素分泌功能显著下降,凋亡细胞比例明显增加,抗凋亡基因Bcl2 m RNA表达水平下调,促凋亡基因Bax m RNA表达水平上调,Bcl2/Bax比例明显降低。结论人胰岛淀粉样多肽上调促凋亡基因表达和抑制抗凋亡基因表达可能与糖尿病的发生发展相关。
Objective To investigate the effects of high glucose on the apoptosis of rat insulinoma cells (INS-1) transfected with human amylin gene and its related gene expression. Methods The insulin levels of INS-1 and INS-1 supernatant (h IAPP / INS-1) transfected with human amylin were detected by enzyme linked immunosorbent assay (ELISA). Hoechst33258 staining The percentage of apoptotic cells was detected by RT-PCR. The levels of Bcl-2 and Bax-m RNA after high glucose stimulation were detected by RTPCR. Results Compared with non-transfected human amylin INS-1 cells, the insulin secretion of h IAPP / INS-1 cells was significantly decreased and the percentage of apoptotic cells was significantly increased. The expression of anti-apoptotic gene Bcl2 mRNA The levels of Bax m RNA were up-regulated and the ratio of Bcl2 / Bax was significantly decreased. Conclusions Upregulation of pro-apoptotic gene expression and inhibition of anti-apoptotic gene expression by human amylin may be related to the development of diabetes mellitus.