目的 研究夏枯草提取物(EPV)对人乳头瘤病毒(HPV)阳性人宫颈癌细胞的凋亡作用。方法 培养人宫颈癌细胞SiHa、HeLa(HPV阳性)及C-33-A(HPV阴性),与20、40、80 μg/mL的EPV共孵育24 h,对照组加入等体积的DMSO,采用噻唑蓝(MTT)法检测细胞存活率;流式细胞术Annexin V/PI双染法检测细胞凋亡及细胞周期。取对数生长期的SiHa、HeLa细胞,与40 μg/mL EPV共孵育24 h,Western blotting法检测凋亡相关蛋白caspase 3、caspase 9、Bcl-2、Bax的表达情况。结果 EPV剂量相关地抑制HPV阳性人宫颈癌细胞SiHa、HeLa活力(P<0.05、0.01、0.001);诱导SiHa、HeLa细胞凋亡(P<0.05、0.01),使SiHa、HeLa细胞周期阻滞在G₀/G₁期,抑制其增殖;对C-33-A细胞无显著作用。显著上调SiHa、HeLa细胞内Cle caspase 3、Cle caspase 9、Bax表达,下调Bcl-2表达(P<0.05、0.01、0.001)。结论 EPV可诱导HPV阳性人宫颈癌SiHa、HeLa细胞凋亡,对HPV阴性C-33-A细胞无显著作用,为夏枯草及其制剂在临床上治疗宫颈癌提供依据。

Objective To study the potential apoptotic effect of extract from Prunella vulgaris (EPV), a traditional Chinese medicine, on human papilloma virus (HPV) positive and negative human cervical cancer cells. Methods Human cervical carcinoma cell lines SiHa, Hela (HPV positive), and C-33-A (HPV negative) were cultured, and treated with 20, 40, and 80 μg/mL of EPV for 24 h. Control group were treated with equal volume of DMSO. The survival rate of cells was detected by methyl thiazolyl tetrazolium (MTT) method. Flow cytometry Annexin V/PI double staining method was used to detect cell apoptosis and cell cycle. SiHa and HeLa cells were incubated with 40 μg/mL EPV for 24 h, and Western blotting was used to detect the expression of apoptosis related protein caspase 3, caspase 9, Bcl-2, and Bax. Results EPV reduced the viability of SiHa and HeLa cells dose-dependently (P<0.05, 0.01 and 0.001). Moreover, EPV induced apoptosis in SiHa and HeLa cells (P<0.05 and 0.01), inhibited the proliferation of SiHa and HeLa cells through arresting their cycle in G₀/G₁ phase. However, EPV did not show significantly effect on C-33-A cells. Western blotting results showed that Cle caspase 3, Cle caspase 9, Bax, and Bcl-2 proteins were involved in EPV induced cell apoptosis (P<0.05, 0.01, and 0.001). Conclusion EPV could induce apoptosis in HPV-positive cervical cancer cells, which provides novel information for clinical application of P. vulgaris and its preparations for the HPV-positive cervical cancer patients.