4-苯基丁酸类似物的设计合成及对PC12细胞内质网应激与过度自噬的抑制作用

Design and synthesis of 4-phenylbutyric acid analogues and their inhibition effect on the endoplasmic reticulum stress and over-autophagy in PC12 cells

目的:设计合成4-苯基丁酸(PBA)类似物并探究其对PC12细胞内质网应激(ERS)和过度自噬的双重抑制作用。方法:以PBA为母核,设计合成了11个PBA类似物(A1-A11),通过1H NMR、13C NMR、MS确证化学结构,采用MTT法分析化合物A1-A11对毒胡萝卜素(TG)诱导的PC12细胞ERS损伤模型和H2O2诱导的PC12细胞过度自噬损伤模型中细胞的保护作用;Western blot法检测ERS的标志蛋白葡萄糖调节蛋白78(GRP78)和自噬标志蛋白LC3 Ⅱ/Ⅰ和Beclin-1表达水平。结果:共合成了11个PBA类似物,其中A3、A7、A9-A11为新化合物。MTT法结果表明,PBA类似物A1对TG诱导的ERS损伤细胞具有保护作用(P<0.05)。A1、A2、A4和A6-A10对H2O2诱导的过度自噬损伤细胞均有良好的保护活性(P<0.05)。Western blot检测结果表明,A1能显著抑制ERS标志性蛋白GRP78、自噬标志性蛋白LC3 Ⅱ/Ⅰ和Beclin-1的表达水平(P<0.05)。结论:与PBA相比,设计合成的化合物A1对ERS和过度自噬的损伤细胞具有最强的双重保护作用。

Objective:To design and synthesize 4-phenylbutyric acid(PBA)analogues and explore their dual inhibition effect on the endoplasmic reticulum stress(ERS)and excessive autophagy in PC12 cells.Methods:Compounds(A1-A11)were designed and synthesized based on the structure of PBA whose chemical structures were confirmed by 1H NMR,13C NMR and MS.MTT was used to analyze the protective effect of A1-A11 on endoplasmic reticulum injury model of PC12 cells induced by TG and injury model of PC12 cells induced by H2O2;Western blot was used to measure the levels of glucose regulated protein 78(GRP78),LC3 Ⅱ/Ⅰ and Beclin-1.Results:Compounds A3,A5,A7 and A9-A11 were new compounds.MTT assay showed that A1 displayed good protective effect on endoplasmic reticulum stress injury model(P<0.05).A1,A2,A4 and A6-A10 indicated great protective effect on autophagy injury model(P<0.05).Western blot showed that A1 could significantly inhibit the expression of GRP78,LC3 Ⅱ/Ⅰ and Beclin-1(P<0.05).Conclusion:Compared with PBA,A1 performs excellent dual protective effect on ERS and autophagy injured model.