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目的:通过抑制磷酸化丝/苏氨酸激酶(phosphorylation serine/threonine kinase,p-Akt)的表达,观察并探讨p-Akt和端粒酶逆转录酶(telomerase reverse transcriptase,TERT)对高糖高脂干预的胰岛β细胞凋亡的影响。方法:将NIT-1细胞随机分成对照组(正常培养)、Akt抑制组(正常培养+Akt抑制剂SH5)、高糖高脂干预组(高糖高脂培养)、高糖高脂+Akt抑制组(高糖高脂培养+Akt抑制剂SH5),TUNEL法和流式细胞术检测各组细胞凋亡,Western blot检测各组细胞TERT、Akt和p-Akt蛋白表达水平,ELISA检测各组细胞胰岛素分泌。结果:与正常对照组比较[凋亡率:(2.88±1.25)%;胰岛素分泌:(0.65±0.10);p-Akt:(0.80±0.07);TERT:(1.63±0.12)];高糖高脂干预组胰岛β细胞凋亡率(38.25±8.63)%明显增加,胰岛素分泌(0.40±0.09)下降,p-Akt(0.55±0.05)和TERT(1.03±0.05)表达明显下降,差异均有统计学意义(P<0.05);与高糖高脂干预组[凋亡率:(38.25±8.63)%;胰岛素分泌:(0.40±0.09);p-Akt:(0.55±0.05);TERT:(1.03±0.05)]比较,高糖高脂+Akt抑制组细胞凋亡率(62.00±7.13)%明显增高,胰岛素分泌(0.23±0.10)下降,p-Akt(0.28±0.07)和TERT(0.61±0.07)表达明显下降,差异均有统计学意义(P<0.05)。结论:抑制p-Akt表达可增加高糖高脂诱导的胰岛β细胞凋亡,其机制可能与降低了TERT表达水平有关。
OBJECTIVE: To observe the effects of p-Akt and telomerase reverse transcriptase (TERT) on the expression of phosphorylation serine / threonine kinase (p-Akt) Effect of Lipid on Islet β Cell Apoptosis. Methods: NIT-1 cells were randomly divided into control group (normal), Akt inhibition group (normal culture + Akt inhibitor SH5), high glucose and high fat intervention group (high glucose and high fat culture), high glucose and lipid + Akt (High glucose and high fat culture + Akt inhibitor SH5), TUNEL method and flow cytometry were used to detect the apoptosis of each group. Western blot was used to detect the expression of TERT, Akt and p-Akt in each group. Insulin secretion. Results: Compared with the normal control group, the apoptotic rate was (2.88 ± 1.25)%; insulin secretion: (0.65 ± 0.10); p-Akt: (0.80 ± 0.07); TERT: (1.63 ± 0.12) The apoptosis rate of pancreatic β-cells (38.25 ± 8.63)% and the secretion of insulin (0.40 ± 0.09) decreased significantly in the lipid-treated group. The expressions of p-Akt (0.55 ± 0.05) and TERT (1.03 ± 0.05) (P <0.05). Compared with the high glucose and high fat intervention group, the apoptosis rate was (38.25 ± 8.63)% and the insulin secretion was (0.40 ± 0.09) (0.23 ± 0.10), p-Akt (0.28 ± 0.07), and TERT (0.61 ± 0.07), respectively. There was no significant difference between the two groups ) Expression was significantly decreased, the differences were statistically significant (P <0.05). Conclusion: Inhibition of p-Akt expression may increase the apoptosis of pancreatic β-cell induced by high glucose and high fat, which may be related to the decrease of TERT expression.