摘要
目的:研究1,8-桉叶油素(1,8-Cineole)对四氧嘧啶诱导胰岛β细胞NIT-1损伤的保护作用及其可能的作用机制。方法:利用四氧嘧啶建立NIT-1细胞损伤模型,并以1.25、2.5、5μmol/L1,8-Cineole预处理30 min。实验分为对照组、模型组和1,8-Cineole高、中、低剂量组。采用MTT法、划痕实验分别检测1,8-Cineole对NIT-1细胞损伤的存活率、划痕修复率的影响;生化法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)含量;ELISA检测细胞上清液中胰岛素的分泌情况;Western Blot检测Bcl-2、Bax和Caspase-9蛋白的表达;机制研究进一步分为对照组、模型组、1,8-Cineole组、Z-VAD-FMK组和1,8-Cineole+Z-VAD-FMK组,采用Western Blot检测Caspase-9蛋白的表达。结果:与对照组比较,模型组细胞存活率、划痕修复率显著降低(P<0.01),上清液中SOD、CAT、GSH-Px含量以及胰岛素分泌量均显著减少(P<0.01),MDA含量显著增加(P<0.01),Bax、Caspase-9蛋白表达显著上调(P<0.01),Bcl-2蛋白表达显著下调(P<0.01)。加入1,8-Cineole预处理后,低、中、高剂量组能显著提高细胞存活率(P<0.05,P<0.01),高、中剂量组能提高划痕修复率(P<0.05),高剂量组SOD、CAT、GSH-Px以及胰岛素的分泌显著增加(P<0.01,P<0.05),MDA含量显著降低(P<0.05),中、高剂量组Bax、Caspase-9蛋白表达显著下调(P<0.05,P<0.01),Bcl-2蛋白表达显著上调(P<0.01)。采用Caspase凋亡抑制剂Z-VAD-FMK作用细胞后,Caspase-9蛋白表达显著下调(P<0.05)。结论:1,8-Cineole能改善四氧嘧啶诱导的NIT-1细胞损伤,其作用机制与改善氧化应激程度和抑制细胞凋亡有关。
Objective:To investigate the protective effect of 1,8-Cineole on pancreatic isletβcells(NIT-1)injuryinduced by Alloxan and its possible mechanism.Methods:NIT-1 cells injury model was made by Alloxan and were pretreated with 1,8-Cineole at different concentrations(1.25,2.5,5μmol/L)for 30 min in vitro.The experiment were divided into control group,Alloxan group,high dose group,middle dose group and low dose group of 1,8-Cineole,separately.The survival rate of 1,8-Cineole on injured NIT-1 cells was detected by MTT assay,and cell repairment rate was analyzed by wound healing method.The contents of antioxidant related factors such as malondialdehyde(MDA),superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)were determined by biochemical method and the secretion of insulin in cell supernatant was determined by enzyme linked immunosorbent assay(ELISA),the expression of Bcl-2,Bax and Caspase-9 proteins were determined by Western Blot.Research on mechanism included control group,Alloxan group,1,8-Cineole group,Z-VAD-FMK group,as well as the combination of 1,8-Cineole and Z-VAD-FMK group,and then Caspase-9 protein was determined by Western Blot.Results:Compared with the control group,the cell survival rate and cell repairment rate were significantly decreased(P<0.01),the content of SOD,CAT,GSH-Px and insulin in cell supernatant decreased significantly(P<0.01),and MDA content increased dramatically(P<0.01).The expression of Bax and Caspase-9 proteins were significantly up regulated(P<0.01)and Bcl-2 protein was obviously down regulated(P<0.01).After pretreated with 1,8-Cineole,the cell survival rate significantly increased in high,middle and low dose group(P<0.05,P<0.01),the cell repairment rate increased in high and middle dose group(P<0.05),the secretion of SOD,CAT,GSH-Px and insulin increased significantly in high dose group(P<0.01,P<0.05)and MDA content decreased significantly(P<0.05)in high dose group,the expression of Bax and Caspase-9 proteins expression were significantly down regulated(P<0.05,P
作者
文波
罗芳
高群
付凌云
陶玲
沈祥春
WEN Bo;UO Fang;GAO Qun;FU Ling-yun;TAO Ling;SHEN Xiang-chun(Key Laboratory of Optimal Utilizaiton of Natural Medicine Resources,Guizhou Medical University,Guiyang 550025,China;Department of Microbiology and Biochemical Pharmacy,Guizhou Medical University,Guiyang 550025,China;School of Pharmacy,Guizhou Medical University,Guiyang 550025,China;Affliated Hospital of Guizhou Medical University,Guiyang 550001,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第4期1839-1843,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81760725)
国家自然科学基金委-贵州喀斯特中心项目子课题(No.U1812403-4-4)
贵州省科技计划项目(No.黔科合基础[2020]1Z069)。
