冬凌草甲素通过调控UBC9表达对肝星状细胞增殖、凋亡的影响

Effects of Oridonin on proliferation and apoptosis in hepatic stellate cells by regulating UBC9

目的探讨冬凌草甲素对肝星状细胞增殖、凋亡的影响和分子机制。方法采用不同浓度的冬凌草甲素干预肝星状细胞(HSC-T6)48 h。分别采用噻唑蓝(MTT)实验、流式细胞术检测HSC-T6细胞增殖和凋亡变化。采用实时荧光定量PCR(RT-qPCR)和蛋白质印记(Western blot)检测泛素结合酶9(UBC9)表达变化。转染UBC9小干扰RNA(si-UBC9)下调HSC-T6细胞中UBC9表达水平,采用上述方法检测细胞增殖、凋亡变化。结果冬凌草甲素干预后,HSC-T6细胞增殖抑制率、凋亡率显著升高,UBC9表达显著降低,并具有显著的剂量依赖关系(P<0.05)。干扰UBC9表达后HSC-T6细胞增殖抑制率、凋亡率显著升高,UBC9表达显著降低(P<0.05)。过表达UBC9能够逆转冬凌草甲素对HSC-T6细胞的增殖、凋亡的影响(P<0.05)。结论冬凌草甲素能够抑制肝星状细胞增殖,诱导细胞凋亡,其机制与下调UBC9表达有关。

Objective To explore the effects of Oridonin on the proliferation and apoptosis of hepatic stellate cells and their molecular mechanism. Methods Different concentrations of Oridonin were used to intervene in hepatic stellate cells( HSC-T6) for 48 h. The methyl thiazolyl tetrazolium( MTT) assay and flow cytometry were used to detect proliferation and apoptosis in HSC-T6 cells. Real-time quantitative polymerase chain reaction( RT-qPCR) and Western blot were used to detect the expression changes of ubiquitin-conjugating enzyme 9( UBC9). The transfection of UBC9 small interfering RNA( si-UBC9) downregulated the expression level of UBC9 in HSC-T6 cells. Changes in cell proliferation and apoptosis were also evaluated using the aforementioned method. Results After Oridonin intervention,the proliferation inhibition rate and apoptosis rate of HSC-T6 cells were significantly increased,the expression of UBC9 was significantly decreased and there was a significant dose-dependent relationship( P<0. 05). After downregulating UBC9 expression,the proliferation inhibition rate and apoptosis rate of HSC-T6 cells were significantly increased and UBC9 expression was significantly decreased( P < 0. 05). The overexpression of UBC9 was able to reverse the effects of Oridonin on the proliferation and apoptosis of HSC-T6 cells( P < 0. 05). Conclusions Oridonin can inhibit proliferation and induce apoptosis in hepatic stellate cells. The underlying mechanism is related to the downregulation of UBC9 expression.