丝胶通过PI3K/Akt信号通路对受损INS-1细胞糖酵解的调控作用

Regulatory Effects of Sericin on Glycolysis of Damaged INS-1 Cells through PI3K/Akt Signal Pathway

摘要

目的 观察丝胶对链脲佐菌素（STZ）致损伤大鼠胰岛素瘤细胞（INS-1细胞）PI3K/Akt信号通路和糖酵解的调控作用。方法 实验以体外培养的INS-1细胞为研究对象随机分为五组,正常对照组、模型组、丝胶组、Akt1抑制剂组、Akt1激动剂组。运用蛋白印迹和实时荧光定量PCR法检测各分组细胞的磷脂酰肌醇-3-激酶（PI3K）,蛋白激酶B（Akt）,磷酸果糖激酶-1（PFK1）,6-磷酸果糖-2,6-二磷酸酶（PFKFB2）的蛋白及mRNA的表达情况。结果 与正常对照组相比,模型组PI3K,p-Akt,PFK1,PFKFB2的蛋白表达显著下降（P<0.05）;丝胶组与模型组相比PI3K,p-Akt,PFK1,PFKFB2的蛋白表达显著增高（P<0.05）;Akt1抑制剂组与丝胶组相比,p-Akt,PFK1,PFKFB2的蛋白表达显著降低（P<0.05）;Akt1激动剂组与丝胶组相比,p-Akt,PFK1,PFKFB2的蛋白呈现上升的趋势。各组INS-1细胞中PI3K,Akt,PFK1,PFKFB2 mRNA的变化趋势与蛋白一致。结论 丝胶对STZ致损伤INS-1细胞发挥保护作用的机制可能是,靶向Akt1影响PI3K/Akt信号通路增强糖酵解。

Abstract

Objective To observe the regulatory effects of sericin on PI3K/Akt signal pathway and glycolysis of INS-1 cells injured by streptozotocin (STZ). Methods INS-1 cells cultured in vitro were randomly divided into five groups. Normal control group, model group, sericin group, Akt1 inhibitor group and Akt1 agonist group. Western blotting and real-time fluorescence quantitative PCR were used to detect the expression of phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), phosphofructosekinase-1 (PFK1), 6-phosphofructose-2,6-diphosphatase (PFKFB2) protein and mRNA. Results Compared with the normal control group, the protein expression of PI3K, p-Akt, PFK1, PFKFB2 of INS-1 cells in the model group decreased significantly (P<0.05). The protein expression of PI3K, p-Akt, PFK1, PFKFB2 of INS-1 cells in the sericin group were significantly higher than that in the model group (P<0.05). The protein expression of p-Akt, PFK1, PFKFB2 of INS-1 cells in the Akt1 inhibitor group were significantly lower than that in the sericin group (P<0.05). Compared with sericin group, the protein expression of p-Akt, PFK1 and PFKFB2 showed an upward trend in the Akt1 agonist group. The change trend of PI3K, Akt, PFK1, PFKFB2 mRNA expression in INS-1 cells of each group were consistent with that of protein. Conclusion The protective mechanism of sericin on STZ-induced injury of INS-1 cells may be that targeted Akt1 affects PI3K/Akt signal pathway and enhances glycolysis.

关键词

丝胶 / 糖尿病 / INS-1细胞 / PI3K/Akt信号通路 / 糖酵解

Key words

sericin / diabetes / INS-1 cells / PI3K/Akt signaling pathway / glycolysis

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基金资助

河北省“三三三人才工程”资助项目（A2016005063）; 承德医学院优势学科B资助