目的 探討京尼平及其衍生物對硝普鈉誘導(dǎo)的PC12細(xì)胞損傷的保護作用及其作用機制。方法 通過62.5~1 000 μmol/L硝普鈉處理PC12細(xì)胞24 h建立細(xì)胞損傷模型，造模前2 h給予京尼平衍生物預(yù)處理，采用噻唑藍(lán)比色法（MTT）考察京尼平衍生物對PC12細(xì)胞存活率的影響。Hoechst染色后觀察細(xì)胞形態(tài)，以DCFH-DA探針和丙二醛（MDA）檢測試劑盒分別檢測PC12細(xì)胞內(nèi)ROS、MDA水平，RT-PCR法檢測PC12細(xì)胞抗氧化應(yīng)激基因水平。結(jié)果 與對照組比較，硝普鈉能劑量相關(guān)性地降低PC12細(xì)胞的存活，在750 μmol/L時具有統(tǒng)計學(xué)差異，而10 μmol/L 1R-異丙基-6，7-二氫京尼平（化合物4）、1S-異丙基-6，7-二氫京尼平（化合物5）能夠明顯增加硝普鈉誘導(dǎo)PC12細(xì)胞損傷后的細(xì)胞存活率（P<0.05、0.01）。Hoechst染色形態(tài)學(xué)證實化合物4、5能夠明顯減少PC12細(xì)胞的凋亡。硝普鈉誘導(dǎo)PC12細(xì)胞內(nèi)氧化應(yīng)激水平，化合物4、5能有效地降低ROS、MDA水平，并促進抗氧化酶基因GCLC、GPX、CAT mRNA表達，但對HO-1、MnSOD基因表達水平無明顯影響。結(jié)論 京尼平衍生物可以抑制硝普鈉誘導(dǎo)的PC12細(xì)胞損傷和細(xì)胞內(nèi)氧化應(yīng)激水平，其作用機制可能是通過影響抗氧化酶GCLC、CAT、GPX mRNA表達水平而實現(xiàn)。

Objective To study the protective effect of genipin derivatives against sodium nitroprusside-induced PC12 cells damage and its mechanisms. Methods PC12 cells were treated with 62.5-1 000 μmol/L sodium nitroprusside for 24 h to evaluate damage, then pre-treated by genipin derivatives for 2 h. Effect of genipin derivatives on cell survival rate of sodium nitroprusside-induced PC12 cells was evaluated by MTT assay. Morphological changes of PC12 cells were observed by Hoechst 33258 nuclear and DCFH-DA staining. ROS and MDA production levels were measured using commercial assay kits. Meantime, the expression of anti-oxidant enzyme gene was determined by RT-PCR. Results Compared with the control group, cell survival rate of sodium nitroprusside-induced PC12 cells decreased in a dose-dependent manner at the dose of 750 μmol/L. Compounds 4 and 5 with concentration of 10 μmol/L significantly decreased the cell apoptosis rate induced by sodium nitroprusside (P<0.05 and 0.01). Hoechst staining showed that compounds 4 and 5 could significantly decrease the apoptosis of PC12 cells. The level of oxidative stress in PC12 cells were induced by sodium nitroprusside. Compounds 4 and 5 could effectively reduce ROS and MDA levels, and promote the mRNA expression of anti-oxidant enzyme genes GCLC, GPX, and CAT, but it showed no significant effect on mRNA levels of HO-1 and MnSOD. Conclusion Genipin derivatives can inhibit level of oxidative stress damage in PC12 cells induced by sodium nitroprusside, which may be related to the change of mRNA expression of anti-oxidant enzyme GCLC, CAT, GPX gene.

國家自然科學(xué)基金資助項目（81560662）；江西省衛(wèi)生計生委中醫(yī)藥科研課題（2015A050）江西中醫(yī)藥大學(xué)博士啟動基金項目（2014BS012）