复方贞术调脂胶囊含药血清对脂毒性胰岛β细胞损伤的干预作用

Intervention effect of Fufang Zhenshu Tiaozhi capsule-medicated serum on lipotoxic injury of isletβcells

目的观察复方贞术调脂胶囊(FTZ)含药血清对棕榈酸(PA)诱导的大鼠胰岛β细胞损伤的干预作用。方法培养大鼠胰岛β细胞INS-1,随机分为空白对照组、牛血清白蛋白(BSA)对照组、PA组、实验组。各组均在RPMI1640完全培养基培养,BSA对照组在培养液中加入1%BSA,PA组、实验组加入0.5 mmol/L的PA溶液,实验组另加入10%FTZ含药血清,各组给药后培养24 h。采用Western blotting法检测细胞中的内质网应激相关蛋白(CHOP)、自噬相关蛋白(ATG5、BECLIN1),采用实时荧光定量PCR法检测CHOP、ATG5、BECLIN1 mRNA,采用DCFH-DA活性氧(ROS)荧光探针检测细胞中的ROS。结果PA组CHOP蛋白、mRNA表达及ROS含量高于空白对照组和BSA对照组,实验组CHOP蛋白、mRNA表达及ROS含量低于PA组(P均<0.05)。PA组ATG5、BECLIN1蛋白和mRNA表达低于空白对照组和BSA对照组,实验组ATG5、BECLIN1蛋白和mRNA表达高于PA组(P均<0.05)。结论FTZ能够减轻氧化应激及内质网应激、增强自噬作用,从而减轻棕榈酸导致的胰岛β细胞损伤。

Objective To observe the intervention effect of serum containing Fufang Zhenshu Tiaozhi capsule(FTZ)on palmitic acid(PA)-induced damage to isletβcells in rats.Methods Rat isletβcells INS-1 were cultured and randomly divided into the blank control group,bovine serum albumin(BSA)control group,PA group,and experimental group,respectively.Cells in all groups were cultured in RPMI1640 complete medium,with the BSA control group receiving 1%BSA in the culture medium,the PA group and experimental group receiving 0.5 mmol/L PA solution,and the experimental group additionally receiving 10%FTZ-containing serum.After treatment,cells were cultured for 24 h.Western blotting was used to detect the expression of endoplasmic reticulum stress-related protein(CHOP)and autophagy-related proteins(ATG5,BECLIN1)in cells,while real-time fluorescence quantitative PCR was used to detect CHOP,ATG5,and BECLIN1 mRNA.The DCFH-DA reactive oxygen species(ROS)fluorescence probe was used to detect ROS in cells.Results The protein and mRNA expression levels of CHOP and ROS content in the PA group were higher than those in the blank control group and BSA control group,while the protein and mRNA levels of CHOP,and ROS content in the experimental group were lower than those in the PA group(all P<0.05).The ATG5 and BECLIN1 protein and mRNA expression levels in the PA group were lower than those in the blank control group and BSA control group,while the ATG5 and BECLIN1 protein and mRNA expression levels in the experimental group were higher than those in the PA group(all P<0.05).Conclusion FTZ can reduce oxidative stress and endoplasmic reticulum stress,enhance autophagy,and thus reduce the PA-induced damage to isletβcells.