论文部分内容阅读
目的探讨18 ku转位蛋白TSPO选择性配体YL-IPA08对皮质酮诱导BV-2细胞凋亡的保护作用,并研究其可能的作用机制。方法选择性TSPO配体YL-IPA08 1~100 nmol·L~(-1)和(或)TSPO拮抗剂PK11195 100 nmol·L~(-1)预处理小胶质细胞系BV-2细胞2 h,加入皮质酮200μmol·L~(-1)继续培养24 h,分别用流式细胞术检测细胞凋亡,CCK-8法检测细胞存活,Western蛋白质印迹法检测TSPO蛋白表达水平,ELISA法检测细胞培养上清中四氢孕酮的水平。结果与溶剂对照组相比,YL-IPA08 1~100 nmol·L~(-1)与阳性化合物AC-5216一样能降低皮质酮处理的BV-2细胞的凋亡率(P<0.01),且100 nmol·L~(-1)时作用最为显著,该作用能被PK11195 100 nmol·L~(-1)所拮抗(P<0.05)。细胞活性检测显示,YL-IPA08 100 nmol·L~(-1)时能显著升高细胞活性(P<0.01),PK11195 100 nmol·L~(-1)处理会拮抗该作用(P<0.01)。Western蛋白质印迹和ELISA法结果显示,YL-IPA08 100 nmol·L~(-1)时能显著增加皮质酮处理的BV-2细胞TSPO蛋白的表达(P<0.05)并显著升高培养液中四氢孕酮的水平(P<0.05),PK11195 100 nmol·L~(-1)显著逆转该现象,降低四氢孕酮的水平(P<0.05)。结论 YL-IPA08显著抑制皮质酮诱导BV-2细胞的凋亡,表现出对小胶质细胞的保护作用,该作用与其增加TSPO蛋白的表达、升高四氢孕酮的水平密切相关。
Objective To investigate the protective effect of 18 ku TSPO selective ligand YL-IPA08 on corticosterone-induced apoptosis of BV-2 cells and its possible mechanism. Methods The microglia BV-2 cells were pretreated with TSPO ligand YL-IPA08 1 ~ 100 nmol·L -1 and / or TSPO antagonist PK11195 100 nmol·L -1 for 2 h , And cultured with 200μmol·L -1 corticosterone for 24 hours. Cell apoptosis was detected by flow cytometry. Cell viability was measured by CCK-8 assay. Western blotting was used to detect the expression of TSPO protein. Culture the level of tetrahydroprotetone in the supernatant. Results Compared with the solvent control group, YL-IPA08 1 ~ 100 nmol·L -1 could reduce the apoptosis rate of corticosterone-treated BV-2 cells (P <0.01) as well as the positive compound AC-5216 The effect of 100 nmol·L -1 was the most significant, which was antagonized by PK11195 100 nmol·L -1 (P <0.05). The cell viability assay showed that 100 nmol·L -1 YL-IPA08 could significantly increase the cell viability (P <0.01). PK11195 100 nmol·L -1 antagonized this effect (P <0.01) . Western blotting and ELISA showed that YL-IPA08 at 100 nmol·L -1 could significantly increase the expression of TSPO protein in corticosterone-treated BV-2 cells (P <0.05) (P <0.05). PK11195 100 nmol·L -1 reversed this phenomenon significantly and decreased the level of tetrahydroprotetone (P <0.05). Conclusion YL-IPA08 significantly inhibits the apoptosis of BV-2 cells induced by corticosterone and shows a protective effect on microglial cells. This effect is closely related to increasing the expression of TSPO protein and increasing the level of tetrahydroprotensone.