淫黄葛合剂诱导HAMP对HT22细胞APP水解中关键酶BACE1表达的影响

The Effect of HAMP Induction by Yinhuangge Mixture on the Expression of BACE1,A Key Enzyme in APP Hydrolysis in HT22 Cells

目的:探索淫黄葛合剂诱导HAMP对HT22细胞β-淀粉样蛋白前体蛋白(Amyloid Precursor Protein,APP)水解中关键酶BACE1表达的影响。方法:CCK-8法检测中药的细胞毒性后,将HT22细胞随机分为正常组、Aβ组、RNAi组、Aβ+RNAi组、Aβ+TCM组、RNAi+TCM组和Aβ+RNAi+TCM组,应用Real-Time PCR检测各组BACE1的表达情况。结果:与正常组比较,Aβ组、RNAi组和Aβ+RNAi组中的BACE1表达增高(P<0.05);与Aβ组比较,Aβ+RNAi组中BACE1表达增高(P<0.05),Aβ+TCM组中BACE1表达降低(P<0.05);与RNAi组比较,Aβ+RNAi组中的BACE1表达增高(P<0.05),RNAi+TCM组中的BACE1表达降低(P<0.05);与Aβ+RNAi组比较,Aβ+RNAi+TCM组中的BACE1表达降低(P<0.05)。结论:淫黄葛合剂可以通过上调HAMP的表达,缓解脑细胞铁超载,抑制BACE1的表达,从而减少APP的淀粉样代谢通路的产生。

Object:To explore the effect of HAMP induction by Yinhuangge mixture on the expression of BACE1,a key enzyme in APP hydrolysis in HT22 cells.Methods:HT22 cells were randomly divided into 7 groups:normal group,Aβgroup,RNAi group,Aβ+RNAi group,Aβ+TCM group,RNAi+TCM group and Aβ+RNAi+TCM group after detecting the cytotoxicity of Chinese medicine by CCK-8 method,and the expression of BACE1 in each group was detected by applying Real-time PCR.Results:Real-time PCR results showed that BACE1 expression was increased in the Aβgroup,RNAi group and Aβ+RNAi group compared with the Control group(P<0.05);BACE1 expression was increased in the Aβ+RNAi group compared with the Aβgroup(P<0.05)and decreased in the Aβ+TCM group(P<0.05);BACE1 expression was increased in the Aβ+RNAi group(P<0.05)and decreased in the RNAi+TCM group(P<0.05)when compared with the RNAi group;BACE1 expression was decreased in the Aβ+RNAi+TCM group(P<0.05)when compared with the Aβ+RNAi group.Conclusion:Yinhuangge mixture can alleviate iron overload in brain cells by upregulating the expression of HAMP,inhibit the expression of BACE1,and reduce the production of amyloid metabolic pathway of APP.