高糖环境下糖络宁对大鼠背根神经节神经元细胞凋亡水平及MAPKs信号通路的影响

Effects of Tangluo Ning on apoptosis and MAPKs signaling pathway of rat dorsal root ganglion neurons in high glucose environment

目的研究糖络宁对高糖环境下大鼠背根神经节神经元(DRGn)细胞凋亡及丝裂原激活的蛋白激酶(MAPK)信号通路的影响。方法6~8周体重(215±15)g清洁级雄性SD大鼠60只,按照随机数字表法分为高、中、低浓度糖络宁组和正常组,每组15只。各组分别以糖络宁组方生药5、2.5、1.25 g/(kg·d)和等量蒸馏水灌胃,制备含药血清和正常对照血清。新生SD胎鼠制备DRGn细胞,分为正常组(10%正常大鼠血清+Neurobasal培养基),高糖组(正常组基础上+50 mmol/L葡萄糖),糖络宁各浓度组(高糖组基础上+10%低、中、高浓度糖络宁含药血清),培养24 h后检测细胞凋亡及JNK1、p-JNK1、c-Jun、p-c-Jun、Map3k8、p-Map3k8蛋白表达。结果高糖组细胞凋亡率、p-JNK1、p-c-Jun、Map3k8、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值高于正常组,差异有统计学意义(P<0.05或P<0.01);糖络宁各浓度组细胞凋亡率、p-JNK1、p-c-Jun、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值明显低于高糖组,差异有高度统计学意义(P<0.01);中浓度糖络宁组细胞凋亡率、p-JNK1、p-c-Jun、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值明显低于高、低浓度糖络宁组,差异有统计学意义(P<0.01或P<0.05);中、低浓度糖络宁组c-Jun表达低于高糖组,差异有统计学意义(P<0.05),糖络宁各浓度组JNK1、Map3k8及c-Jun表达比较,差异无统计学意义(P>0.05)。结论糖络宁可能通过抑制MAPKs信号通路降低细胞凋亡水平。

Objective To study the effect of Tangluo Ning on apoptosis and mitogen-activated protein kinases(MAPKs)signaling pathway of rat dorsal root ganglion neurons(DRGn)in high glucose environment.Methods Sixty clean grade male SD rats with body weight of(215±15)g at 6-8 weeks were divided into high,medium,and low concentration glucolonin groups and normal group according to random number table method,with 15 rats in each group.Each group was given 5,2.5,1.25 g/(kg·d)crude medicine of Tangluo Ning prescription and equal amount of distilled water intragastric administration to prepare drug-containing serum and normal control serum.DRGn cells were prepared from newborn SD fetal rats and were divided into normal group(10%normal rat serum+Neurobasal medium),high glucose group(based on normal group+50 mmol/L glucose),and Tangluo Ning concentration groups(based on high glucose group+10%low,medium,and high concentration Tangluo Ning drug-containing serum).Cell apoptosis and protein expression of JNK1,p-JNK1,c-Jun,p-c-Jun,Map3k8,and p-Map3k8 were detected after 24 h culture.Results The apoptosis rate,expression of p-JNK1,p-c-Jun,Map3k8,and p-Map3k8 and ratio of p-JNK1/JNK1,p-c-Jun/c-Jun,and p-Map3k8/Map3k8 in high glucose group were higher than those in normal group,the differences were statistically significant(P<0.05 or P<0.01).The apoptosis rate,expression of p-JNK1,p-c-Jun,and p-Map3k8 and ratio of p-JNK1/JNK1,p-c-Jun/c-Jun,and p-Map3k8/Map3k8 in Tangluo Ning concentration groups were significantly lower than those in high glucose group,the differences were highly statistically significant(P<0.01).Apoptosis rate,expression of p-JNK1,p-c-Jun,and p-Map3k8 and ratio of p-JNK1/JNK1,p-c-Jun/c-Jun,and p-Map3k8/Map3k8 in middle concentration Tangluo Ning group were significantly lower than those in high and low concentration Tangluo Ning group,the differences were statistically significant(P<0.01 or P<0.05).The expression of c-Jun in the medium and low concentration Tangluo Ning groups was lower than that in the high concentration Tangluo Ning group,and the differences were statistically significant(P<0.05).There was no significant difference in the expression of JNK1,Map3k8 and c-Jun in the Tangluo Ning concentration groups(P>0.05).Conclusion Tangluo Ning can reduce apoptosis by inhibiting MAPKs signaling pathway.