山绿茶中Ilexgenin A诱导宫颈癌HeLa细胞凋亡及其相关机制研究

The study on apoptosis of human cervical cancer HeLa cell induced by Ilexgenin A from Ilex hainanensis Merr and its mechanism

目的研究山绿茶中的乌苏烷型三萜化合物Ilexgenin A对人宫颈癌HeLa细胞凋亡的诱导作用及其相关机制。方法建立人宫颈癌HeLa细胞体外培养体系,采用CCK-8法检测细胞活力,MTT法测定LDH,免疫比色法检测细胞增殖情况,DAPI荧光染色观察细胞凋亡形态学变化,Western blot检测Caspase-3/7和NF-κB蛋白表达水平。结果用IA处理过的HeLa细胞,LDH释放增加,呈剂量和时间依赖性。在CCK-8和BrdU测定过程中,IA比标准药物抗HeLa活性更强,随浓度和作用时间的增加,细胞的活性不断下降,对HeLa增殖的抑制作用明显。DAPI染色观察用IA处理过的HeLa细胞浓缩明显,细胞核分散,染色质凝聚。Western blot结果显示用IA处理过的HeLa细胞Caspase-3/7的活性显著增加,NF-κB蛋白表达水平下降。结论 Ilexgenin A具有诱导HeLa细胞凋亡的作用,其机制可能与激活Caspase-3/7和抑制NF-κB蛋白表达有关。

Objective To investigate the apoptosis of human cervical cancer HeLa cell induced by Ilexgenin A from llex hain- anensis Merr and its mechanism. Methods The viability of HeLa cells was detected by the method CCK - 8. The activity of LDH was measured by MTF assay. Immune colorimetry was used to test the proliferation of Hela cells. The change of morphology of ap- optosis HeLa cells was observed by DAPI fluorescence staining. Western blot was used to detect the expressions of Caspase - 3/7 and NF - κB proteins of HeLa cells. Results After HeLa cells were treated by IA, the leakage of LDH increased in a dosage and time dependent manner. In the process of determination of BrdU and CCK - 8, the inhibitory effect of IA to HeLa cell was stronger than standard drugs, and showed clear inhibitory effect on the HeLa cell proliferation with the increasing of concentration and the acting time. By using the DAPI fluorescence dyes, the HeLa cell cytoplasm concentrating, nucleolus dispersion and chromatin ag- glomeration were observed. The Western blot analysis showed that the expression of Caspase - 3/7 was significantly enhanced and the expression of NF - xB proteins was down - regulated. Conclusion Ilexgenin A could induce the apoptosis of HeLa cells and its mechanism related to activating the activity of Caspase - 3/7 and down - regulating the expression of NF - κB protein.