筋脉通含药血清对高糖培养的Schwann细胞诱导型一氧化氮合酶和NADPH氧化酶p22-phox亚基表达的影响

Effects of Chinese herbal medicine Jinmaitong-medicated serum on inducible nitric oxide synthase and NADPH oxidase p22-phox subunit of rat Schwann cells cultured in high-glucose medium

目的:探讨筋脉通含药血清对高糖培养的Schwann细胞诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)蛋白表达及烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotidephosphate,NADPH)氧化酶p22-phox亚基mRNA表达的影响。方法:雄性Wistar大鼠随机分为3组,分别灌胃筋脉通、维生素C或蒸馏水制备含药血清和对照血清。取新出生大鼠的双侧坐骨神经用于制备Schwann细胞。将体外培养的Schwann细胞分为高糖组、筋脉通组(加入筋脉通含药血清)、维生素C组(加入维生素C含药血清)及正常对照组。培养48h后,采用免疫荧光法检测Schwann细胞内i NOS蛋白的表达,实时荧光定量聚合酶链式反应法检测NADPH氧化酶p22-phox亚基mRNA的表达。结果:高糖培养的Schwann细胞内i NOS蛋白表达及NADPH氧化酶p22-phox亚基mRNA表达均较正常对照组明显升高(P<0.01);与高糖组比较,筋脉通组i NOS蛋白表达及NADPH氧化酶p22-phox亚基mRNA表达明显降低(P<0.01),且明显优于维生素C组(P<0.01)。结论:筋脉通含药血清可以下调高糖培养的Schwann细胞i NOS蛋白及NADPH氧化酶p22-phox亚基mRNA的表达。

Objective：To investigate the effects of medicated serum prepared by administration of Jinmaitong（JMT）,a compound Chinese herbal medicine,on nicotinamide adenine dinucleotide phosphate（NADPH）oxidase p22-phox subunit and inducible nitric oxide synthase（iNOS）of rat Schwann cells cultured in high-glucose medium.Methods：Wistar rats were divided into normal control group（distilled water）,JMT（JMT at dose of 1.31 g/（kg·d））group and vitamin C（vitamin C at dose of 0.08 g/（kg·d））group to prepare medicated serum.Bilateral sciatic nerves of new born Wistar rats were used to separate Schwann cells.Schwann cells cultured in high-glucose medium were divided into high glucose group（cultured with 50 mmol/L glucose medium）,JMT group（cultured with JMT-medicated serum）and vitamin C（VC）group（cultured with VC-medicated serum）.Schwann cells cultured in DMEM were used as the normal control.After 48 h culturing,the expression of iNOS was detected by immunofluorescence method and p22-phox mRNA was tested by real-time fluorescent quantitative polymerase chain reaction.Results：The expression levels of iNOS and p22-phox mRNA in the high glucose group were higher than those in the normal control group（P0.01）.Compared with the high glucose group,expressions of iNOS protein and p22-phox mRNA in JMT group were significantly decreased（P0.01）and JMT-medicated serum had better effect than VC（P0.01）.Conclusion：JMT-medicated serum can down-regulate the expressions of iNOS protein and NADPH oxidase p22-phox subunit mRNA of Schwann cells cultured in high-glucose medium.