HSP70的表达在不同预处理下星形孢菌素诱导的大鼠神经细胞毒性应激损伤模型中的意义

摘要

目的评估不同预处理对抑制星形孢菌素（STS）诱导的神经细胞毒性应激损伤的作用。方法选取 SD大鼠乳鼠120只，分为对照组（n =20）、STS 模型组（n =20）、姜黄素组（n =20）、槲皮素+ STS 模型组（n =20）、姜黄素+STS 预处理组（n =20）、姜黄素+槲皮素+ STS 处理组（n =20）6个组。取乳鼠海马神经细胞，利用 STS 诱导建立神经细胞毒性应激损伤模型。对照组研究期间不作任何特殊处理，STS 模型组、槲皮素+ STS 模型组、姜黄素+ STS 预处理组、姜黄素+槲皮素+ STS 处理组、姜黄素组分别加入终浓度为20μmol/ L 的 STS、终浓度为10μmol/ L 槲皮素+20μmol/ L ；STS、终浓度为20μmol/ L 的姜黄素和槲皮素、终浓度为10μmol/ L 的槲皮素+20μmol/ L 的姜黄素和槲皮素、终浓度为20μmol/ L 的姜黄素。利用噻唑蓝（MTT）法、乳酸脱氢酶（LDH）释放率和 WesternBlot 法分别测定各组细胞存活率、细胞毒性损伤程度和 HSP70表达量。结果姜黄素+ STS 预处理组、姜黄素组细胞存活率明显高于 STS 模型组，差异具有统计学意义（ P ＜0.01）；姜黄素+ STS 预处理组和姜黄素组神经细胞毒性率明显低于 STS 模型组，差异有统计学意义（ P ＜0.01）；姜黄素+ STS 预处理组 HSP70表达量高于 STS 模型组和姜黄素组，差异有统计学意义（ P ＜0.01）；姜黄素+ STS 模型组神经元存活率高于槲皮素+ STS 预处理组、姜黄素+槲皮素+ STS 预处理组和 STS 模型组，差异具有统计学意义（ P ＜0.05）。结论姜黄素能增强 HSP70的表达并抑制 STS 诱导的 SD 乳鼠神经元细胞毒性损伤，而槲皮素通过阻断 HSP70表达使姜黄素的抑制作用被抵消。%Objective To evaluate the effect of different preconditioning on the inhibition of staurosporine(STS)- induced neurotoxicity stress injury. Methods 120 SD suckling rats were divided into 6 groups:control group,STS model group,curcumin group,quercetin + STS model group,curcumin + STS pretreatment group and curcumin + quercetin + STS treatment group. Curcumin was added to inhibit STS - induced neuronal injury and the expression of HSP70 was blocked by quercetin. The control group did not make any special treatment during the study peri-od,STS model group,quercetin + STS model group,curcumin + STS pretreatment group,curcumin + quercetin + STS treatment group and curcu-min group were added with final concentration of 20 μmol/ L STS,final concentration of 10 quercetin + 20 μmol / L STS,20 μmol/ L curcumin and quercetin at the final concentration,the final concentration of 10 μmol/ L quercetin + 20 μmol/ L of curcumin and quercetin,final concentration of 20 μmol/ L curcumin. MTT assay was used to detect the expression of HSP70 in the neurons of hippocampal neurons. Lactate dehydrogenase (LDH)release rate and Western Blot were used to determine cell viability,cytotoxicity and HSP70 expression. Results The survival rate of cur-cumin + STS group and curcumin group was significantly higher than that of STS model group( P < 0. 01). The rate of neurotoxicity was signifi-cantly lower than that of STS model group( P < 0. 01). Expression of HSP70 in curcumin + STS pretreatment group was higher than STS model group and curcumin group( P < 0. 01). Curcumin + STS model group was higher than quercetin + STS pretreatment group,curcumin + quercetin + STS pretreatment group and STS model group,the difference was statistically significant( P <0. 05). Conclusion Curcumin can enhance the expression of HSP70 and inhibit STS - induced neurotoxicity stress injury,while quercetin can inhibit the expression of HSP70 in nerve cells,and counteract the inhibitory effect of curcumin.