白藜芦醇通过SIRT1/NF-κB信号通路对热化疗诱导人结肠上皮细胞的保护作用

Protective Effects of Resveratrol on Human Colon Epithelial Cells Induced by Hyperthermic Chemotherapy Through SIRT1/NF-κB Signaling Pathway

目的:研究沉默信息调节因子1(silent information regulator 1,SIRT1)信号通路在白藜芦醇(resveratrol,Res)改善热化疗(hyperthermic chemotherapy,HTC)诱导人结肠上皮细胞(NCM460)损伤中的作用。方法:体外传代培养NCM460细胞,将其分为对照组、HTC组、HTC+低剂量(5μg/mL)Res组、HTC+中剂量(10μg/mL)Res组、HTC+高剂量(20μg/mL)Res组,采用10 mg/L顺铂结合43℃温热法干预NCM460细胞30 min,采用CCK8法分析NCM460细胞的相对存活率;流式细胞仪检测NCM460细胞的凋亡率;Western Blot法检测SIRT1/NF-κB通路蛋白及凋亡相关蛋白Bax、MDM2的表达。结果:与对照组比较,HTC组NCM460细胞相对存活率降低、细胞凋亡率升高(P<0.05),SIRT1蛋白表达降低,p-NF-κB、Bax、MDM2蛋白表达升高(P<0.05);与HTC组比较,低、高剂量Res组NCM460细胞相对存活率增高、细胞凋亡率降低(P<0.05),SIRT1蛋白表达升高,p-NF-κB、Bax、MDM2蛋白表达降低(P<0.05)。结论:Res对NCM460细胞的HTC损伤具有改善作用,其作用机制可能与抑制SIRT1/NF-κB信号通路活化有关。

Objective:To study the effects of silent information regulator 1(SIRT1)signaling pathway on resveratrol(Res)improving the injury of human colon epithelial cells(NCM460)induced by hyperthermic chemo-therapy(HTC).Methods:NCM460 cells were subcultured in vitro,and they were divided into the control group,HTC group,HTC+low dose(5μg/mL)of Res group,HTC+moderate dose(10μg/mL)of Res group and HTC+high dose(20μg/mL)of Res group,after treating NCM460 cells with hyperthermia at 43℃and cisplatin at 10 mg/L for 30 minutes,CCK8 method was used to analyze relative survival rate of NCM460 cells;flow cytometry to detect the apoptosis rate of NCM460 cells;Western blot method to survey the expressions of SIRT1/NF-κB pathway protein and apoptosis-related protein Bax and MDM2.Results:Compared with the control group,relative survival rate of NCM640 cells in HTC group reduced,cellular apoptosis rate rose(P<0.05),the expressions of SIRT1 protein lowered,the expressions of p-NF-κB,Bax and MDM2 protein rose(P<0.05);compared with HTC group,relative survival rate of NCM460 cells in low and high doses group of Res rose,cellular apoptosis rate reduced(P<0.05),the expressions of SIRT1 protein lifted,the expressions of p-NF-κB,Bax and MDM2 protein reduced(P<0.05).Conclusion:Res possesses protective effects on HTC damage of NCM460 cells,and its mechanism might be related to inhibiting the activation of SIRT1/NF-κB signaling pathway.