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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 study the induction of Ilex Genin A, a ursane-type triterpene in human green tea, on the apoptosis of human cervical carcinoma HeLa cells and its related mechanisms. Methods The human cervical cancer HeLa cells were cultured in vitro. Cell viability was detected by CCK-8 assay. LDH was measured by MTT assay. Cell proliferation was detected by immunochromatography. Morphological changes of apoptotic cells were observed by DAPI staining. Caspase- 3/7 and NF-κB protein expression levels. Results HeLa cells treated with IA showed an increase in LDH release in a dose and time dependent manner. In CCK-8 and BrdU assay, IAA was more active than standard drugs in anti-HeLa activity. With the increase of concentration and duration of action, the activity of IAA decreased continuously and the inhibitory effect on HeLa proliferation was obvious. DAPI staining observed with IA-treated HeLa cells concentrated significantly, nuclear dispersion, chromatin condensation. Western blot results showed that the activity of Caspase-3/7 in HeLa cells treated with IA significantly increased and the expression of NF-κB decreased. Conclusion Ilexgenin A can induce the apoptosis of HeLa cells. The mechanism may be related to the activation of Caspase-3/7 and the inhibition of NF-κB protein expression.