筋脉通含药血清对高糖培养SD大鼠背根神经元NADPH氧化酶p47^(phox)亚基及NT表达的影响

Effects of Jinmaitong medicated serum on the expression of NADPH oxidase p47^(phox) subunit and nitrotyrosine in dorsal root ganglion neurons cultured in high glucose medium

目的:探讨筋脉通含药血清对高糖培养大鼠背根神经节神经元(DRGn)NADPH氧化酶p47^(phox)亚基及NT表达的影响。方法:雄性SD大鼠随机分为3组,即筋脉通组、牛磺酸组和对照组,分别灌胃筋脉通、牛磺酸、蒸馏水制备含药血清和对照血清。从孕15d的SD胎鼠背根神经节取材制备细胞悬液。将体外培养的DRGn分为高糖组、筋脉通组(加入筋脉通含药血清)、牛磺酸组(加入牛磺酸含药血清)及对照组,培养24h后,CCK-8法检测细胞活性。采用免疫荧光和Western blot检测NADPH氧化酶p47^(phox)亚基及NT的蛋白表达。结果:与对照组比较,高糖组DRGn细胞活性显著降低,NADPH氧化酶p47^(phox)亚基及NT蛋白表达水平显著升高(P<0.01);与高糖组比较,筋脉通组和牛磺酸组DRGn细胞活性显著升高,NADPH氧化酶p47^(phox)亚基及NT蛋白表达水平显著降低(P<0.01)。结论:筋脉通可降低高糖培养DRGn NADPH氧化酶p47^(phox)亚基及NT的表达,减轻氧化应激反应,其作用与牛磺酸含药血清相当。

Objective:To investigate the effects of Jinmaitong(JMT)on the expressions of NADPH oxidase p47phox subunit and nitrotyrosine(NT)in dorsal root ganglion neurons(DRGn)cultured in high glucose medium.Methods:Male SpragueDawley(SD)rats were randomly divided into control group(distilled water),JMT group(JMT at dose of 1.31 g·kg-1·d-1)and Taurine group(Taurine at dose of 1.2 g·kg-1·d-1)to prepare medicated serum.DRGn were harvested from embryonic 15 days SD rats to prepare cell suspension.DRGn were divided into control group,high glucose(HG)group(cultured with 45 mmol/L glucose medium),JMT group(cultured with JMT containing serum of 10%)and Taurine group(cultured with Taurine containing serum of 10%)were plated for 24 h.Cell viability was detected by CCK-8 colorimetric assay to determine the time point of detection.The protein expressions of NADPH of oxidase p47phox subunit and NT were detected by fluorescence-immunocytochemistry and Western blot.Results:Cell viability decreased significantly and the protein levels of NADPH oxidase p47phox subunit and NT in HG group were much higher than control group(P<0.01).Compared with HG group,cell viability increased significantly and the protein levels of p47phox subunit and NT in both treatment groups remarkably decreased(P<0.01).Conclusion:JMT treatment significantly ameliorated the alteration in expressions of NADPH oxidase p47phox subunit and NT of DRGn cultured in high glucose medium,and its effects is equivalent to taurine.