筋脉通对高糖培养雪旺细胞炎性因子及神经营养因子的影响

Effects of Jinmaitong on the inflammatory cytokines and neurotrophic factors in Schwann cells cultured in high-glucose medium

目的探讨中药筋脉通(JMT)对高糖条件下雪旺细胞(SCs)炎性因子及神经营养因子蛋白表达的影响。方法将雄性SD大鼠随机分为3组,分别以筋脉通胶囊、神经妥乐平(Ntp)及等体积蒸馏水灌胃制备含药血清和正常大鼠血清;原代培养SCs,分为正常组、高糖组、JMT组和Ntp组,48 h后用CCK-8法检测各组SCs细胞增殖,免疫印迹法检测各组SCs细胞IL-1β、IL-6、IL-10、TNF-α、NGF及NT-3蛋白的表达,夹心法酶联免疫吸附法测定各组SCs上清液中IL-6、TNF-α、NGF的表达。结果与正常组比较,高糖组SCs增殖活性减低(P<0.01),TNF-α蛋白表达上调、分泌量增加(P<0.05),IL-1β、L-10蛋白表达上调(P<0.05,P<0.01),NGF的蛋白表达下调、分泌量减少(P<0.05,P<0.01),NT-3蛋白表达下调(P<0.05)。与高糖组比较,JMT组细胞增殖活性提高(P<0.01),TNF-α蛋白表达量及分泌量减少(P<0.05),IL-1β、L-10蛋白表达量减少(P<0.01),NGF蛋白表达量、分泌量及NT-3蛋白表达增加(P<0.05);JMT组SCs细胞增殖活性提高,SCs中TNF-α表达及分泌受到抑制,IL-1β表达量减少,NGF蛋白表达增加,其作用优于Ntp组(P>0.05,P<0.01)。IL-6蛋白表达和分泌在各组间差异无统计学意义(P>0.05)。结论 JMT含药血清能够提高高糖培养SCs的增殖活性,降低TNF-α、IL-1β、L-10等炎性因子表达,抑制TNF-α分泌,提高NT-3表达,促进SCs分泌NGF,从而起到防治DPN的作用。

Objective To investigate the effects of Chinese herbal medicine Jinmaitong（ JMT） on the expression of inflammatory cytokines and neurotrophic factors in Schwann cells（ SCs） exposed to high-glucose medium. Methods Male SD rats were randomly divided into three groups respectively treated by JMT capsule,Neurotropin（ Ntp） and distilled water of equal volume by gavage in order to prepare the medicated serum and normal rat serum. Primary culture SCs was divided into normal group,high glucose group,JMT group and Ntp group. After 48 h culturing, the SCs proliferation of different groups was determined by CCK-8 assay,the expression of IL-1β,IL-6,IL-10,TNF-α,NGF and NT-3 of SCs of all groups was detected by immunoblotting and the expression of IL-6,TNF-α,NGF in the supernatant of SCs of all groups by enzyme-linked immunosorbent assay（ ELISA）. Results Compared with the normal group,the cell proliferation viability of high-glucose group was reduced significantly（ P〈0. 01）,and the expression and secretion of TNF-α（ P〈0. 05）,the expression of IL-1β,IL-10（ P〈0. 05,P〈0. 01） were markedly increased,and the expression and secretion of NGF（ P〈0. 05,P〈0. 01）,and the expression of NT-3（ P〈0. 05） were down-regulated obviously（ P〈0. 05）.Compared with the high-glucose group,the cell proliferation viability of JMT group was increased（ P〈0. 01）,the expression and secretion of TNF-α were reduced markedly（ P〈0. 05）,the expression of IL-1β and L-10（ P〈0. 01） were decreased obviously,the expression and secretion of NGF and the expression of NT-3 were all increased remarkably（ P〈0. 05）. The cell proliferation viability of SCs of JMT group was significantly increased,the inhibition of the expression of TNF-α in SCs cells,the secretion of TNF-α,and the expression of IL-1β and the increase of the expression of NGF in JMT group were all better than that of Ntp group 改 为 the expression and secretion of TNF-α were inhibited,the expression of IL-β was decreased,the expression of NGF of SCs was increased,the actions were better than that of Ntp group（ P〈0. 05,P〈0. 01）. Conclusion The medicated serum of JMT can strengthen the proliferation viability of SCs cells,and lower the expression of inflammatory cytokines,like TNF-α,IL-1β and L-10,inhibit the secretion of TNF-α,increase the expression of NT-3,and promote the secretion of NGF in SCs cells,thus preventing and treating diabetic peripheral neuropathy（ DPN）.