摘要
目的观察益糖康含药血清对IR-HepG2细胞葡萄糖消耗的影响,并探讨其可能的作用机制。方法采用高胰岛素刺激建立稳定可靠的IR-HepG2细胞模型;益糖康高、中、低血清、空白血清、二甲双胍血清分别干预IR-HepG2细胞,葡萄糖过氧化物酶法测定IR-HepG2细胞葡萄糖消耗量;MTT法检测细胞活性;Annexin V—FITC/PI法测定益糖康含药血清对IR-HepG2细胞凋亡的影响;RT-PCR法检测IR-HepG2细胞IRS-2、PI3K、AKT基因的表达;Western Blot法检测IR-HepG2细胞IRS-2蛋白及AKT磷酸化蛋白的表达。结果1×10^(-6) mol/L胰岛素作用24h可建立稳定IR-HepG2细胞模型;当益糖康含药血清浓度15%,作用时间48h时,葡萄糖消耗量到达最大;与模型组相比较,益糖康含药血清能够降低凋亡率,抑制细胞凋亡;提高IRS-2、PI3K、AKT基因的表达(P<0.01),提高IRS-2、p-AKT蛋白的表达(P<0.01)。结论15%益糖康含药血清、作用48h使IR-HepG2葡萄糖消耗量达到最大,抑制IR-HepG2凋亡,其作用机制可能与上调IRS-2/PI3K/AKT信号通路中关键分子的基因和蛋白表达有关。
Objective To observe the effect of Yitangkang serum on glucose consumption of IR-HepG2 cells and explore its possible mechanism.Methods a stable and reliable IR-HepG2 cell model was established by high insulin stimulation;yitangkang high,medium,low serum,blank serum and metformin serum were used to interfere with IR-HepG2 cells,glucose consumption of IR-HepG2 cells was measured by glucose peroxidase method;cell activity was measured by MTT method;annexin was used the expression of IRS-2,PI3K and AKT in IR-HepG2 cells was detected by RT-PCR,and the expression of IRS-2 and AKT in IR-HepG2 cells was detected by Western Blot.Results 1×10^(-6)mol/L insulin for 24 hours to establish a stable IR-HepG2 cell model;when the serum concentration of Yitangkang was 15%and the action time was 48 hours,the glucose consumption reached the maximum;compared with the model group,Yitangkang serum could reduce the apoptosis rate and inhibit the apoptosis;compared with the model group,Yitangkang serum could improve the expression of IRS-2,PI3K and AKT genes;the expression of IRS-2 and p-AKT protein was increased(P<0.01).Conclusion 15%Yitangkang serum can increase the glucose consumption of IR-HepG2 and inhibit the apoptosis of IR-HepG2.The mechanism may be related to the up regulation of gene and protein expression of key molecules in IRS-2/PI3K/AKT signaling pathway.
作者
薛玲
石岩
XUE Ling;SHI Yan(Liaoning University of Traditional Chinese Medicine College of traditional Chinese Medicine,Shenyang 100847,China;Liaoning University of Traditional Chinese Medicine,Shenyang 100847,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2022年第10期2374-2377,共4页
Lishizhen Medicine and Materia Medica Research
基金
2019年辽宁中医药大学校内高层次人才引进科研启动项目
国家重点基础研究发展计划(973计划)(2013CB532004)
国家科技支撑计划项目(2014BAI10B04)。
