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目的:观察木瓜三萜对吲哚美辛诱导RGM-1细胞凋亡的影响,并探讨其可能的机制。方法:用吲哚美辛诱导RGM-1细胞建立细胞损伤模型,MTT法检测不同浓度木瓜三萜对RGM-1细胞增殖、流式细胞术和Hoechst33258染色分析木瓜三萜对吲哚美辛诱导细胞凋亡的影响,JC-1染色法测定细胞线粒体跨膜电位水平,Caspase试剂盒检测RGM-1细胞中Caspase-9和Caspase-3的活性,Real-time PCR检测RGM-1细胞中TFF1、p53、Bcl-2及Bax mRNA表达,Western blot检测RGM-1细胞中TFF1和p53蛋白表达。结果:当木瓜三萜浓度低于100μg/m L时,木瓜三萜对RGM-1细胞的生长无明显的影响;当木瓜三萜浓度低于50μg/m L时,木瓜三萜与吲哚美辛联用对RGM-1细胞的生长有促进作用;木瓜三萜可显著增加吲哚美辛诱导RGM-1细胞损伤的活性(P<0.05,P<0.01),升高线粒体膜电位,抑制细胞凋亡和改善细胞形态,上调TFF1蛋白表达及TFF1、Bcl-2 mRNA表达和Bcl-2与Bax的比值,下调p53蛋白表达及p53和Bax mRNA表达,降低Caspase-9及Caspase-3活性(P<0.05,P<0.01)。结论:木瓜三萜可有效抑制吲哚美辛诱导RGM-1细胞凋亡,其作用机制可能与上调TFF1、Bcl-2,下调p53、Bax、Caspase-3、Caspase-9表达有关。
Objective: To observe the effect of papaya triterpenoids on indomethacin-induced apoptosis in RGM-1 cells and to explore its possible mechanism. Methods: Cell injury model was induced by indomethacin in RGM-1 cells. MTT assay was used to detect the proliferation of RGM-1 cells by different concentrations of papaya triterpenes. Flow cytometry and Hoechst33258 staining were used to analyze the effect of papaya triterpene on indomethacin-induced cells The Caspase kit assay Caspase-9 and Caspase-3 activity in RGM-1 cells, Real-time PCR detection of RGC-1 cells TFF1, p53 , Bcl-2 and Bax mRNA expression were detected by Western blot. The expression of TFF1 and p53 in RGM-1 cells was detected by Western blot. Results: When the papaya triterpenoids concentration was less than 100μg / m L, the papaya triterpenes had no significant effect on the growth of RGM-1 cells. When the papaya triterpenoids concentration was less than 50μg / m L, the triterpene and indomethacin Syndrom was effective on the growth of RGM-1 cells; papaya triterpenoid could significantly increase the indomethacin-induced RGM-1 cell injury activity (P <0.05, P <0.01), increased mitochondrial membrane potential and inhibited cells Apoptosis and cell morphology, TFF1 protein expression, TFF1 and Bcl-2 mRNA expression, Bcl-2 and Bax ratio, p53 protein and Bax mRNA expression and the activity of Caspase-3 <0.05, P <0.01). CONCLUSION: The papaya triterpene can effectively inhibit indomethacin-induced apoptosis of RGM-1 cells. The mechanism may be related to the up-regulation of TFF1 and Bcl-2, down-regulation of p53, Bax, Caspase-3 and Caspase-9 expression.