雷公藤红素抑制Cd2+诱导的神经毒性

Celastrol inhibits neurotoxicity induced by Cd2+

镉(cadmium,Cd)是环境中常见的一种重金属,Cd^(2+)可以通过穿透血脑屏障,产生神经毒性,从而诱发各种神经退行性疾病,雷公藤红素是雷公藤的一种有效成分,具有抗癌、抗炎等一系列药理作用,本文探究雷公藤红素对Cd^(2+)诱导的相应神经毒性的影响作用。通过细胞增殖实验、细胞膜完整性实验、细胞形态实验探索了Cd^(2+)对小胶质细胞HMC3活力的影响;通过一氧化氮(NO)检测实验、脂质过氧化(malondialdehyde,MDA)检测实验、蛋白免疫印迹实验分析了Cd^(2+)的神经毒性以及雷公藤红素对Cd^(2+)诱导的相应神经毒性的影响。结果表明:与对照组相比,当Cd^(2+)浓度达到40μmol/L时,对HMC3细胞增殖抑制率为(57.17±8.23)%(P<0.01,n=5),继续增大Cd^(2+)浓度,细胞活性将进一步降低;当Cd^(2+)浓度达到40μmol/L以上时,HMC3的细胞膜明显受到破坏,并且破坏作用与浓度呈剂量依赖性关系;随着Cd^(2+)浓度的增加,细胞形态开始变化,贴壁效果变差。Cd^(2+)使HMC3细胞释放的NO量显著增加,而雷公藤红素能够有效地抑制Cd^(2+)诱导的HMC3细胞NO的释放;Cd^(2+)使HMC3细胞脂质过氧化水平显著增加,加入10^(-7) mol/L雷公藤红素后,MDA的释放量显著减少;Cd^(2+)会使p-PI3K蛋白含量增加,而加入了雷公藤红素(10^(-7)、10^(-6) mol/L)后,p-PI3K蛋白和p-AKT蛋白的激活均被抑制,从而抑制了细胞凋亡。综上所述,雷公藤红素能够抑制Cd^(2+)诱导的小胶质细胞毒性,从而起到神经保护作用。

Cadmium(Cd)is a common heavy metal in the environment.Cd^(2+)may penetrate the blood-brain barrier and produce neurotoxicity,thus inducing various neurodegenerative diseases.Celastrol is an effective component of Tripterygium wilfordii Hook.F.,which has many pharmacological effects such as anti-cancer and anti-inflammatory.Here we explored the effect of celastrol on the corresponding neurotoxicity induced by Cd^(2+).Cell proliferation test,cell membrane integrity test,and cell morphology were observed to analyze the effect of Cd^(2+)on the viability of HMC3.The neurotoxicity of Cd^(2+)and the effect of celastrol on the corresponding neurotoxicity induced by Cd^(2+)were analyzed by nitric oxide(NO)test,lipid peroxidation(MDA)test,and Western blotting.When the concentration of Cd^(2+)reached 40μmol/L,the inhibition rate of HMC3 cell proliferation was(57.17±8.23)%(P<0.01,n=5),compared with the control group.The cell activity continued to reduce when the Cd^(2+)concentration further increased.When the concentration of Cd^(2+)was higher than 40μmol/L,the cell membrane of HMC3 was significantly damaged,and the damage was dose-dependent.Upon increasing the Cd^(2+)concentration,the cell morphology began to change and the adhesion also became worse.Cd^(2+)significantly increased the amount of NO released by HMC3 cells,while celastrol effectively inhibited the NO release of HMC3 cells induced by Cd^(2+).Cd^(2+)greatly increased the release of MDA in HMC3 cells,and the level of MDA decreased rapidly upon the addition of 10^(-7) mol/L celastrol.Cd^(2+)increased the expression of p-PI3K protein,and the levels of p-PI3K protein and p-AKT protein were inhibited by the addition of celastrol(10^(-7) mol/L,10^(-6) mol/L),thus preventing cell apoptosis.In conclusion,celastrol inhibits Cd^(2+)induced microglial cytotoxicity and plays a neuroprotective role.