目的 研究積雪草酸（AA）對(duì)前列腺癌PC-3細(xì)胞作用及分子機(jī)制。方法 采用CCK8法檢測積雪草酸（5、10、20、40、80、160μmol·L^-1）對(duì)PC-3細(xì)胞活力的影響；采用Hochest法和Annexin V/PI法檢測積雪草酸（20、30、40μmol·L^-1）對(duì)PC-3細(xì)胞凋亡的影響；采用JC-1法檢測積雪草酸對(duì)PC-3細(xì)胞線粒體膜電位的影響；采用Western blotting法檢測積雪草酸對(duì)PC-3細(xì)胞線粒體凋亡途徑和Janus激酶2（JAK2）/信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄活化因子3（STAT3）信號(hào)通路相關(guān)蛋白表達(dá)水平的影響。采用Annexin V/PI法、JC-1法、Western blotting法檢測JAK2激動(dòng)劑coumermycin A1（C-A1）對(duì)積雪草酸誘導(dǎo)PC-3細(xì)胞線粒體凋亡及相關(guān)蛋白表達(dá)的影響。結(jié)果 與對(duì)照組比較，積雪草酸可濃度及時(shí)間相關(guān)性地抑制PC-3細(xì)胞增殖，20～160μmol·L^-1組差異顯著（P＜0.01），作用24、48、72h后的半數(shù)抑制濃度（IC₅₀）值分別為29.98、23.04、13.81μmol·L^-1；20、30、40μmol·L^-1積雪草酸顯著促進(jìn)PC-3細(xì)胞凋亡（P＜0.05、0.01）；可明顯導(dǎo)致PC-3細(xì)胞線粒體膜電位下降；顯著上調(diào)PC-3細(xì)胞線粒體凋亡相關(guān)Bax、cleaved Caspase-3的蛋白表達(dá)（P＜0.05、0.01），顯著下調(diào)Bcl-2的蛋白表達(dá)（P＜0.05、0.01），顯著抑制JAK2/STAT3信號(hào)通路JAK2和STAT3的磷酸化水平（P＜0.05、0.01）。與積雪草酸組比較，積雪草酸＋C-A1顯著抑制PC-3細(xì)胞的凋亡水平（P＜0.01），明顯抑制PC-3細(xì)胞線粒體膜電位下降，顯著下調(diào)Bax、cleaved Caspase-3蛋白表達(dá)（P＜0.01），顯著上調(diào)Bcl-2蛋白表達(dá)（P＜0.01），顯著提升JAK2和STAT3的磷酸化水平（P＜0.01）。結(jié)論 積雪草酸可通過抑制JAK2/STAT3信號(hào)通路誘導(dǎo)前列腺癌細(xì)胞發(fā)生線粒體凋亡，進(jìn)而發(fā)揮抗前列腺癌作用。

Objective To study the effects and molecular mechanism of asiatic acid (AA) on prostate cancer PC-3 cells. Method CCK8 assay was used to detect the effect of AA (5, 10, 20, 40, 80, and 160 μmol·L^-1) on PC-3 cell proliferation. Hochest assay and Annexin V/PI assay were used to detect the effect of AA (20, 30, and 40 μmol·L^-1) on PC-3 cell apoptosis. JC-1 method was used to detect the effect of AA on mitochondrial membrane potential of PC-3 cells. Western blotting was used to detect the effect of AA on the expression level of mitochondrial apoptosis pathway and JAK2/STAT3 signaling pathway related proteins in PC-3 cells. Annexin V/PI method, JC-1 method and Western blotting method were used to detect the effects of JAK2 agonist Coumermycin A1 (C-A1) on mitochondrial apoptosis and expression of related proteins in PC-3 cells induced by AA. Results Compared with control group, AA significantly inhibited the proliferation of PC-3 cells in a concentration-dependent and time-dependent manner, the difference of 20—160 μmol·L^-1 group was significant (P<0.01). and the IC₅₀ value was 29.98, 23.04 and 13.81 μmol·L^-1 after 24, 48 and 72 h. AA significantly induced the apoptosis of PC-3 cells compared with control group (P<0.05 and 0.01). AA also changed the MMP in PC-3 cells. AA significantly up-regulated mitochondrial apoptosis-related Bax and cleaved Caspase 3 expression compared with control group (P<0.05 and 0.01), and significantly down-regulated mitochondrial apoptosis-related Bcl-2 expression (P<0.05 and 0.01). The phosphorylation levels of JAK2 and STAT3 in JAK2/STAT3 signaling pathway were also significantly inhibited by AA (P<0.05 and 0.01) compared with control group. Compared with AA group, AA+C-A1 significantly inhibited the apoptosis of PC-3 cells (P<0.01), inhibited the MMP in PC-3 cells significantly, significantly down-regulated Bax and cleaved Caspase 3 expression in PC-3 cells (P<0.01), and significantly up-regulated Bcl-2 expression (P<0.01), as well as significantly increased the phosphorylation levels of JAK2 and STAT3 (P<0.01). Conclusion AA could induce mitochondrial apoptosis in prostate cancer cells by inhibiting JAK2/STAT3 signaling pathway, and finally play an anti-prostate cancer role.

R285.5

江蘇省自然科學(xué)基金項(xiàng)目（BK20191498）；國家中醫(yī)管理局名醫(yī)驗(yàn)方評(píng)價(jià)與轉(zhuǎn)化重點(diǎn)研究室開放課題（NZYJDMF-2020001）；江蘇省衛(wèi)生健康委醫(yī)學(xué)科研項(xiàng)目重點(diǎn)項(xiàng)目（ZDB2020020）