摘要
目的 探讨丝胶是否通过靶向Akt1调控磷酸肌醇-3-激酶(PI3K)/蛋白激酶B(protein kinase B,Akt)信号通路促进链脲佐菌素(STZ)致损伤大鼠胰岛素瘤细胞(INS-1细胞)增殖.方法 INS-1细胞随机分为4组:对照组(不予任何处理)、模型组(给予10mmol/LSTZ处理细胞)、丝胶组(给予10mmol/LSTZ+600μg/mL丝胶处理细胞)、抑制剂组(给予10mmol/LSTZ+600μg/mL丝胶+0.3 mmol/L的Akt1抑制剂A-674563处理细胞).药物作用时间均为24h.CCK-8法检测各组INS-1细胞存活率.免疫蛋白印迹法(western blot)检测PI3K/Akt信号通路相关蛋白PI3K、p-Akt和增殖相关蛋白增殖细胞核抗原(PCNA)、细胞核增殖相关抗原Ki67(Ki67)的表达情况.结果 与对照组相比,模型组INS-1细胞存活率下降(P＜0.05),PI3K、p-Akt1、PCNA和Ki67的蛋白表达显著下降(P＜0.05);与模型组相比,丝胶组INS-1细胞存活率上升(P＜0.05),PI3K、p-Akt1、PCNA和Ki67的蛋白表达显著增多(P＜0.05);与丝胶组相比,抑制剂组INS-1细胞存活率下降(P＜0.05),p-Akt1、PCNA和Ki67的蛋白表达显著下降(P＜0.05).结论 丝胶具有促进STZ致损伤INS-1细胞增殖的作用,其作用机制与丝胶通过靶向Akt1调控PI3K/Akt信号通路有关.
Abstract
Objective To investigate whether sericin promotes proliferation of streptozotocin (STZ) damaged insulinoma cells (INS-1 cells) through targeting Akt1 regulates PI3K/Akt signaling pathway.Methods INS-1 cells were randomly divided into four groups.In control group,INS-1 cells were cultured under conventional conditions without other treatments.In model group,INS-1 cells were cultured with 10mmol/L STZ.In sericin group,INS-1 cells were cultured with 10mmol/L STZ and 600μg/mL sericin.In inhibitor group,INS-1 cells were cultured with 10mmol/L STZ,600μg/mL sericin and 0.3mmol/L Akt1 inhibitor A-674563.The cells in four groups were cultured with corresponding drugs respectively for 24h.The survival rate of INS-1 cells in each group was detected by CCK-8 method.Western blot was used to detect the expression of PI3K/Akt signaling pathway related proteins PI3K and p-Akt,and proliferation related proteins PCNA and Ki67.Results The survival rate,and the protein expressions of PI3K,p-Akt1,PCNA,and Ki67 of INS-1 cells in model group significantly decreased compared with control group (P＜0.05).The survival rate,and the protein expressions of PI3K,p-Akt1,PCNA,and Ki67 of INS-1 cells in sericin group significantly increased compared with model group (P＜0.05).The survival rate,and the protein expressions of PI3K,p-Akt1,PCNA,and Ki67 of INS-1 cells in inhibitor group significantly decreased compared with sericin group (P＜0.05).Conclusion Sericin can protect proliferation of INS-1 cells damaged by STZ,and the protective mechanisms may related to target Akt1 regulates PI3K/Akt signaling pathway.
基金项目
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()
维普
万方数据