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目的用转染APP695cDNA的SH-SY5Y细胞模型,研究tenuigenin对β-淀粉样蛋白(Aβ)生成的抑制作用及其作用机制。方法应用免疫沉淀和蛋白印迹法(Westernblot)检测Aβ含量,应用MTT和LDH法测定tenuigenin的细胞毒性。结果Tenuigenin降低SH-SY5YAPP695细胞Aβ分泌水平;无细胞增生和毒性作用;能抑制SH-SY5YAPP695细胞APPC99的生成,但对SH-SY5YSPA4CT细胞APPC99生成无抑制作用,亦不影响该细胞株Aβ的分泌。结论Tenuigenin可能通过抑制β-分泌酶对APP的水解,降低SH-SY5YAPP695细胞Aβ水平。Tenuigenin这一针对阿尔茨海默病(Alzheimerdisease,AD)上游病理环节的药效,显示了其在AD治疗方面重要的开发和临床应用的价值,值得进一步研究。
Objective To study the inhibitory effect of tenuigenin on the formation of β-amyloid (Aβ) and its mechanism by SH-SY5Y cell line transfected with APP695 cDNA. Methods Aβ was detected by immunoprecipitation and Western blotting. The cytotoxicity of tenuigenin was determined by MTT assay and LDH assay. Results Tenuigenin decreased the Aβ secretion in SH-SY5YAPP695 cells. No cell proliferation and toxicity were observed. Tenuigenin inhibited the APPC99 production in SH-SY5YAPP695 cells, but did not inhibit the APPC99 production in SH-SY5YAPA4CT cells and did not affect the secretion of Aβ in SH-SY5YAPP695 cells. Conclusion Tenuigenin may reduce the level of Aβ in SH-SY5YAPP695 cells by inhibiting the hydrolysis of APP by β-secretase. The efficacy of Tenuigenin, which is directed against the upper pathological stage of Alzheimerdisease (AD), has shown its value in the development and clinical application of AD therapy and deserves further study.