黄芪甲苷Ⅳ抑制多巴胺诱导的帕金森病SH-SY5Y细胞凋亡的相关机制

Mechanism underlying apoptosis of SH-SY5Y cells in PD induced by astragalosideⅣ-inhibited dopamine

目的探讨黄芪甲苷Ⅳ对神经毒素多巴胺诱导的帕金森病(PD)模型SH-SY5Y细胞凋亡的影响及相关机制。方法取SH-SY5Y细胞株进行培养至对数生长期分为6组:对照组(正常培养)、模型组(多巴胺诱导)、低剂量组(多巴胺+10 mg/ml黄芪甲苷Ⅳ)、中剂量组(多巴胺+30 mg/ml黄芪甲苷Ⅳ)、高剂量组(多巴胺+50 mg/ml黄芪甲苷Ⅳ)与酪氨酸蛋白激酶(JAK)抑制组(多巴胺+30 mg/ml黄芪甲苷Ⅳ+30μmol/L AG490),比较各组细胞存活率、细胞凋亡率、活性氧、JAK2、信号转导子和转录激活子3(STAT3)mRNA、磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)蛋白表达。结果与对照组比较,模型组、低、中剂量组及JAK抑制组细胞存活率明显降低(P<0.05),其他5组细胞凋亡率和活性氧水平更高,JAK2和STAT3 mRNA、p-JAK2和p-STAT3蛋白表达更低(P<0.05);与模型组比较,中、高剂量组细胞存活率、JAK2和STAT3 mRNA、p-JAK2及p-STAT3蛋白表达更高,低、中、高剂量组细胞凋亡率和活性氧水平更低(P<0.05)。低、中、高剂量组JAK2和STAT3 mRNA、p-JAK2及p-STAT3蛋白表达逐渐升高(0.50±0.03 vs 0.66±0.02 vs 0.81±0.03,0.33±0.02 vs 0.56±0.03 vs 0.77±0.05,0.24±0.03 vs 0.42±0.02 vs 0.55±0.04,0.36±0.03 vs 0.56±0.03 vs 0.73±0.04,P<0.05);JAK抑制组JAK2和STAT3 mRNA、p-JAK2及p-STAT3蛋白表达明显低于模型组(P<0.05)。结论黄芪甲苷Ⅳ能抑制多巴胺诱导的PD模型SH-SY5Y细胞凋亡及氧化应激水平,可能与JAK2/STAT3信号通路上调有关。

Objective To study the effect of astragalosideⅣon apoptosis of SH-SY5Y cells in PD model induced by dopamine and its mechanism.Methods SH-SY5Y cells were divided into control group(cultured in normal culture),model group(induced by dopamine),low dose group(cultured in dopamine+10 mg/ml astragalosideⅣ),medium dose group(cultured in dopamine+30 mg/ml astragalosideⅣ),high dose group(cultured in dopamine+50 mg/ml astragalosideⅣ)and JAK inhibition group(cultured in dopamine+30 mg/ml astragalosideⅣ+30μmol/L AG490).The survival rate,apoptosis rate and active oxygen level of SH-SY5Y cells,and expressions of JAK2 and STAT3 mRNA,p-JAK2 and p-STAT3 protein in different groups were compared.Results The survival rate of SH-SY5Y cells was significantly lower in model group,low dose group,medium dose group and JAK inhibition group than in control group(P<0.05)while the apoptosis rate and active oxygen level of SH-SY5Y cells were significantly higher and the expression levels of JAK2 and STAT3 mRNA,p-JAK2 and p-STAT3 protein were significantly lower in model group,low dose group,medium dose group,high dose group and JAK inhibition group than in control group(P<0.05).The survival rate of SH-SY5Y cells and expression levels of JAK2 and STAT3 mRNA,p-JAK2 and p-STAT3 protein were significantly higher in medium dose group and high dose group than in model group while the survival rate and active oxygen level of SH-SY5Y cells were significantly lower in low dose group,medium dose group and high dose group than in model group(P<0.05).The expression levels of JAK2 and STAT3 mRNA,p-JAK2 and p-STAT3 protein increased gradually in low dose group,medium dose group and high dose group(0.50±0.03 vs 0.66±0.02 vs 0.81±0.03,0.33±0.02 vs 0.56±0.03 vs 0.77±0.05,0.24±0.03 vs 0.42±0.02 vs 0.55±0.04,0.36±0.03 vs 0.56±0.03 vs 0.73±0.04,P<0.05)and were significantly lower in JAK inhibition group than in model group(P<0.05).Conclusion Astragaloside IV can inhibit dopamine-induced apoptosis and oxidative stress level of SH-SY5Y cells in PD model by upregulating the JAK/STAT3 signaling pathway.