大豆异黄酮对阿特拉津损伤SH-SY5Y细胞的保护作用及其机制

Mechanism of isoflavones protection of SH-SY5Y cells from atrazine

目的:比较大豆异黄酮、槲皮素、茶多酚、姜黄素和白藜芦醇5种植物化合物对阿特拉津(ATR)损伤神经元的保护作用及分子机制。方法:体外培养SH-SY5Y细胞,分别加入浓度为5、10、20、50、100μmol/L的5种植物化合物,24 h后,加入250μmol/L ATR,继续培养24 h后,CCK-8法检测细胞活力,Annexin V-FTTC/PI双染检测细胞凋亡情况,DCFH-DA探针检测活性氧(ROS)水平,Western blot检测酪氨酸氢化酶(TH)蛋白表达量,并用SPSSS 20.0进行统计学分析。结果:5种植物化合物在低剂量范围内抗ATR损伤效果显著,剂量分别为大豆异黄酮5μmol/L、白藜芦醇5μmol/L、槲皮素50μmol/L、姜黄素5μmol/L、茶多酚20μmol/L,与其他4种植物化合物相比,大豆异黄酮抗ATR损伤SH-SY5Y细胞效果显著,尤其在5μmol/L,可显著降低ROS水平,增加TH蛋白的表达(P<0.05)。结论:大豆异黄酮可能通过降低SH-SY5Y细胞内ROS水平,增加TH蛋白的表达,增加细胞活力,抑制细胞凋亡。

OBJECTIVE： We tested the ability of five candidate compounds-isoflavones,resveratrol,quercetin,curcumin,and green tea polyphenols-to protect SH-SY5Y cells from atrazine （ATR） via in vitro experiments. METHODS：The cells were incubated for 24 h in the presence of various doses （5,10,20,50,and 100 μmol/L） of isoflavones,resveratrol,quercetin,curcumin,or green tea polyphenols. Cells were then cultured for 24 h with 250 μmol/L ATR. At the indicated time after treatment,the CCK-8 method was used to test for cell viability. Apoptosis was detected by using Annexin V-FITC and PI double dye,oxygen radical level was detected by using DCFH-DA method,and the protein expression of tyrosine hydroxylase （TH） by using Western blot. RESULTS：Five plant compounds showed significant anti-ATR effect at low doses,isoflavones 5 μmol/L,resveratrol 5 μmol/L,quercetin 50 μmol/L,curcumin 5 μmol/L and green tea polyphenols 20 μmol/L. Compared with other four plant compounds,the effect of isoflavones anti-ATR injury on SH-SY5Y was significant,especially in 5 μmol/L （P〈0.05）. And isoflavones （5 μmol/L） could significantly reduce the level of ROS and increase the expression of TH （P〈0.05）. CONCLUSION：Isoflavones could protect SH-SY5Y cells from ATR by reducing intracellular ROS levels,increasing the expression of TH and cell viability to inhibit cell apoptosis.