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目的观察姜黄素干预对肝星状细胞(HSC)中髓样分化因子88(My D88)蛋白表达及HSC凋亡的影响。方法将常规传代培养的HSC分为对照组、My D88高表达组、My D88阻断剂组和姜黄素组。My D88高表达组将构建的重组原核表达质粒p ET-32a-p My D88转染至HSC;转染72 h后,My D88阻断剂组加入氨基噻唑类My D88特异性抑制剂继续作用24 h,姜黄素组加入姜黄素继续作用24 h。收集各组细胞,采用流式细胞术检测My D88蛋白的相对表达量[以荧光指数(FI)表示],采用CCK-8法检测各组细胞凋亡率。结果流式细胞术结果显示,My D88表达水平(FI值)在对照组、My D88高表达组、My D88阻断剂组和姜黄素组组间比较差异有统计学意义(6.10±0.37,14.10±0.23,7.50±0.65,8.60±0.53,P<0.05),其中My D88高表达组、My D88阻断剂组和姜黄素组相对表达量明显高于对照组,My D88阻断剂组和姜黄素组的相对表达量低于My D88高表达组。CCK-8法结果显示,对照组、My D88高表达组、My D88阻断剂组、姜黄素组细胞凋亡率组间比较差异有统计学意义[(23.6±1.3)%、(56.8±2.9)%、(33.4±1.1)%、(35.8±2.2)%,P<0.01],其中My D88高表达组、My D88阻断剂组和姜黄素组HSC细胞的凋亡率明显高于对照组,My D88阻断剂组和姜黄素组的HSC细胞的凋亡率低于My D88高表达组。结论姜黄素和氨基噻唑类My D88特异性抑制剂均可显著下调HSC细胞My D88蛋白表达,并降低HSC细胞的凋亡率。
Objective To observe the effects of curcumin on the expression of myeloid differentiation factor 88 (My D88) and the apoptosis of HSC in hepatic stellate cells (HSC). Methods HSCs in routine subculture were divided into control group, My D88 high expression group, My D88 blocker group and curcumin group. My D88 high expression group will be constructed recombinant prokaryotic expression plasmid pET-32a-p My D88 transfected to HSC; 72 h after transfection, My D88 blocker group with aminothiazole My D88 specific inhibitor continued role 24 h, curcumin group added curcumin continued role 24 h. The cells in each group were collected and the relative expression of My D88 protein was detected by flow cytometry [expressed as fluorescence index (FI)]. The apoptosis rate of each group was detected by CCK-8. Results The results of flow cytometry showed that the expression level of My D88 was significantly different between control group, My D88 high expression group, My D88 blocker group and curcumin group (6.10 ± 0.37,14.10 ± 0.23,7.50 ± 0.65,8.60 ± 0.53, P <0.05). The relative expression level of My D88 high-expression group, My D88 blocker group and curcumin group was significantly higher than that of control group, My D88 blocker group and curcumin The relative expression level of the hormone group was lower than that of the My D88 high expression group. The results of CCK-8 assay showed that there was significant difference between the control group, My D88 high expression group, My D88 blocker group and curcumin group [(23.6 ± 1.3)%, (56.8 ± 2.9) ), (33.4 ± 1.1)%, (35.8 ± 2.2)%, P <0.01]. The apoptosis rates of HSC cells in My D88 overexpression group, My D88 blocker group and curcumin group were significantly higher than those in control group , The apoptosis rate of HSC cells in My D88 blocker and curcumin groups was lower than that in My D88 high expression group. Conclusion Both curcumin and aminothiazole-specific inhibitors of My D88 can significantly down-regulate the expression of My D88 protein in HSC cells and decrease the apoptosis rate of HSC cells.