中药筋脉通含药血清对高糖培养雪旺细胞NF-κB表达的影响

Effects of Serum Containing Jinmaitong Capsule on NF-κB of Schwann Cells Cultured with High Glucose Medium

目的观察中药筋脉通含药血清对高糖培养雪旺细胞(SCs)中NF-κB蛋白及其mR-NA表达的影响。方法原代培养并纯化W istar乳鼠坐骨神经SCs,蛋白S-100免疫组织化学法进行鉴定。取第3代SCs,加入不同培养基,分为正常组、高糖组、筋脉通组和神经妥乐平组进行培养,48h后采用激光扫描共聚焦显微技术检测NF-κB蛋白的表达,实时荧光定量PCR技术检测NF-κBmRNA的表达。结果正常组NF-κB绿色荧光强度较弱,主要表达于胞浆区;高糖组荧光强度于胞浆区和胞核区表达都较正常组增强;筋脉通组和神经妥乐平组表达都较高糖组减弱,主要表达于胞浆区。高糖组SCs中NF-κB的蛋白及其mRNA表达较正常组明显增强(P<0.01);与高糖组比较,筋脉通组和神经妥乐平组NF-κB的蛋白及其mRNA的表达显著下降(P<0.01)。结论筋脉通含药血清可抑制高糖培养SCs中NF-κB蛋白及其mRNA的表达。

Objective To investigate the effects of serum containing jinmaitong capsule on nuclear factor kappa B（NF-κB）of Schwann cells（SCs）cultured with high glucose medium as well as the impacts on its mRNA expression.Methods SCs were primarily-cultured and purified from the sciatic nerves of newborn Wistar rats.S-100 protein immunohistochemical method was adopted for the identification of them.SCs of the 3rd passage were collected and cultured with different mediums.They were divided into a normal group,a high-glucose group,a jinmaitong group and a neurotropin group.48h later,confocal laser scanning microscopy was adopted to detect the expression of NF-κB;and real-time fluorescence quantitative PCR was used to detect the expression of NF-κB mRNA.Results In normal group,the green fluorescence intensity of NF-κB was weakly expressed in cytoplasm.The expressions of the green fluorescence intensity in cytoplasm and nuclei were stronger in high-glucose group than that in normal group.The expression in jinmaitong group and neurotropin group were weakened as compared with high-glucose group,mainly in cytoplasm.The expressions of NF-κB protein and its mRNA in high-glucose group were enhanced remarkably as compared with normal group（P0.01）.Compared with high-glucose group,the expressions of NF-κB protein and its mRNA in Jinmaitong group and neurotropin group decreased significantly（P0.01）.Conclusion Serum containing jinmaitong capsule inhibits the expressions of NF-κB and its mRNA cultured with high glucose medium.