目的 研究香葉木苷對(duì)人肝癌HepG2細(xì)胞增殖抑制及誘導(dǎo)凋亡的作用，并探討其相關(guān)分子機(jī)制。方法 將HepG2細(xì)胞分為對(duì)照組和香葉木苷組，香葉木苷組分別加入終質(zhì)量濃度為1、5、25 μg/mL的香葉木苷DMSO溶液，對(duì)照組加入等體積的DMSO。藥物處理24 h后，臺(tái)盼藍(lán)染色及Live/Dead染色檢測(cè)香葉木苷對(duì)HepG2細(xì)胞活性的影響；CCK-8及EdU染色檢測(cè)香葉木苷對(duì)HepG2細(xì)胞增殖的影響；TUNEL染色檢測(cè)香葉木苷對(duì)HepG2細(xì)胞凋亡的影響；實(shí)時(shí)熒光定量PCR（qRTPCR）及Western blotting法檢測(cè)香葉木苷對(duì)Bcl-2、Bax mRNA和蛋白水平的影響。結(jié)果 與對(duì)照組比較，香葉木苷降低HepG2細(xì)胞活力、抑制細(xì)胞增殖，5、25 μg/mL組差異顯著（P<0.05、0.01、0.001）；促進(jìn)HepG2細(xì)胞凋亡，各濃度組均差異顯著（P<0.01、0.001），且均呈劑量相關(guān)性。經(jīng)5 μg/mL香葉木苷處理后，HepG2細(xì)胞中抗凋亡蛋白Bcl-2的mRNA水平顯著降低（P<0.001），而促凋亡蛋白Bax的mRNA水平顯著升高（P<0.001），Bcl-2/Bax蛋白水平顯著降低（P<0.001）。結(jié)論 香葉木苷抑制HepG2細(xì)胞活力和增殖、促進(jìn)凋亡，作用機(jī)制與調(diào)控Bcl-2/Bax表達(dá)有關(guān)。

Objective To study the effects of diosmin on proliferation inhibition and apoptosis of human hepatoma HepG2 cells and to explore its molecular mechanism. Methods HepG2 cells were divided into control group and diosmin group. Diosmin group was added DMSO solution with the final concentration of 1, 5 and 25 μg/mL, and control group was added DMSO with the same volume. After treatment for 24 h,Trypan blue staining and Live/Dead staining were performed to test the effect of diosmin on the cell viability. CCK-8 assay and EdU staining were applied to test the influence of diosmin on the proliferation of HepG2. TUNEL staining was performed to detectthe effect of diosmin on apoptosis of HepG2. QRT-PCR and western blotting were applied to test the expressions of apoptotic-related proteins in each group. Results Compared with the control group, diosmin decreased the cell viability and inhibited the proliferation of HepG2 cells, and there were significant differences between 5 and 25 μg/mL groups (P<0.05, 0.01 and 0.001), and promoted the apoptosis of HepG2 cells in all concentration groups (P<0.01 and 0.001), all were dose related. After 5 μg/mL diosmin treatment, the expression of anti-apoptotic protein Bcl-2 mRNA in HepG2 cells decreased significantly (P<0.001), while the expression of pro-apoptotic protein Bax mRNA increased significantly (P<0.001). The level of Bcl-2/Bax protein decreased significantly (P<0.001). Conclusion Diosmin could decrease cell viability and proliferation, as well as promote apoptosis of HepG2 by regulating Bcl-2/Bax pathway. These findings may represent a new idea for drug therapy of liver cancer.

黑龍江省自然科學(xué)基金項(xiàng)目（H2015044，H2015045）；國(guó)家自然科學(xué)基金項(xiàng)目（81274034）