目的建立钙蛋白酶抑制剂MDL-28170对丙烯酰胺(ACR)诱发神经干细胞(NSCs)毒性的干预模型。方法取Wistar新生大鼠的脊髓背根神经节进行体外细胞培养,采用细胞免疫化学染色检测细胞的巢蛋白(nestin)、神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)以进行NSCs鉴定。将处于对数生长期的NSCs分为对照(培养液)组和ACR单独染毒(140、350、560、700μmol/L)组及MDL-28170干预(700μmol/LACR+65、130、195、260μmol/L MDL-28170)组,暴露48h后,采用MTT法测定细胞存活率。结果细胞的nestin、NSE及GFAP鉴定结果均呈阳性。与对照组相比,仅700μmol/LACR染毒组NSCs的存活率较低,各剂量MDL-28170干预组NSCs的存活率均较低,差异有统计学意义(P〈0.05);且随着ACR染毒剂量的升高,NSCs的存活率呈下降趋势。与700μmol/L ACR染毒组相比,65、130μmol/L MDL-28170干预组NSCs的存活率均较高,而260μmol/L MDL-28170干预组NSCs的存活率较低,差异有统计学意义(P〈0.05)。结论成功建立钙蛋白酶抑制剂对ACR诱发NSCs毒性的干预模型。
Objective To establish the model of intervention effects of calpain inhibitor (MDL-28170) on neural stem cells (NSCs) injury induced by acrylamide. Methods The dorsal root ganglion of postnatal Wistar rats were collected for primary cellular culture. Specific protein nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) in cells were identified by immunocytochemistry staining. The NSCs in logarithmic phase were divided into control group(culture), ACR-infected groups (140,350,560,700 p, mol/L) and intervention groups (700 μmol/L ACR+65,130,195,260 μmol/L MDL-28170). The cells were cultured for 48 h in 5% C02 and at 37 ℃. MTT method was used to determine the cell survival rate. Results Compared with the control group,the cell survival rate in 700 μmol/L ACR group and intervention group were significantly lower (P〈0.05). With the increase of ACR exposure doses,the survival rate of NSCs showed a declining curve. Compared with 700 μmol/L ACR group, the cell survival rates of 65,130 μmol/L intervention groups were significantly higher(P〈0.05), and the cell survival rate of 260 μmol/L intervention group was significantly lower (P〈0.05). Conclusion The model of the intervention effects of calpain inhibitor on NSCs injury induced by acrylamide is established successfully in the study.