降糖通络汤对DPN大鼠NGF mRNA表达的影响及对雪旺细胞抗氧化、抗凋亡作用机制研究

Mechanism study of Jiangtang Tongluo Decoction on NGF mRNA expression in DPN rats and anti-oxidation,anti-apoptotic of rat schwann cells

目的:观察降糖通络汤对实验性糖尿病大鼠周围神经病变(DPN)治疗作用,并通过促进NGF、抗氧化、抗凋亡的环节对其作用机制予以研究。方法:采用静脉注射链脲佐菌素60mg/kg制备DPN模型,按血糖随机分为模型对照组、降糖通络汤36、18、9g生药/kg组、盐酸二甲双胍+甲钴胺分散片对照组(细胞实验为金芪降糖片),给药8周,1次/d。观察给药后2、8周空腹血糖及光热辐射痛阈变化,免疫组织化学测定坐骨神经的神经生长因子(NGF)蛋白表达、RT-PCR法测定NGF m RNA表达。IF测定高糖培养SC细胞中诱导型一氧化氮合成酶(i NOS)表达、RT-PCR法测定高糖培养雪旺细胞还原型辅酶Ⅱ(NADPH)氧化酶p22-phox亚基表达。RT-PCR法测定细胞Casepase-3 m RNA、Bcl-2 m RNA表达,TUNEL法测定雪旺细胞凋亡。结果:降糖通络汤36、18g生药/kg组可明显降低空腹血糖(P<0.01),缩短对热光源刺激的躲避潜伏期(P<0.01)。促进坐骨神经NGF m RNA表达(P<0.01),抑制细胞核i NOS、NADPH p22-phox m RNA表达(P<0.01)、抑制Casepase-3 m RNA表达(P<0.01)、促进Bcl-2 m RNA表达(P<0.01),抑制凋亡率。结论:降糖通络汤通过促进NGF、抑制氧化应激、控制凋亡发挥对周围神经病变的治疗作用。

Objective：To observe the effects of Jiangtang Tongluo Decoction（JTTL） on DPN rats and to study its mechanism through studies of NGF expression,anti-oxidation,anti-apoptotic effects.Methods：DPN rats were made by streptozotocin60mg/kg intravenous.Rats were randomized to 5 groups：DPN model group,JTTL 36,18,9g/kg groups,metformin ＋ mecobalamin tablet（Jin Qi Melbine in cell experiments）.Rats were gavage once a day for 8 weeks.Plasma glucose and pain threshold were determined after 2,8 weeks.Sciatic NGF mRNA expression was tested by RT-PCR,iNOS expression was tested by IF,SC p22-phox,Casepase-3,Bcl-2 mRNA expression was tested by RT-PCR and SC apoptosis was tested by TUNEL.Results：Plasma glucose of 36,18g/kg were significantly decreased（P〈0.01）,pain threshold were significantly shortened（P〈0.01）,NGF mRNA,Bcl-2 mRNA were significantly increased（P〈0.01）,and iNOS,p22-phox,Casepase-3 mRNA were significantly decreased（P〈0.01）,compared to the DPN model rats.Conclusion：JTTL can promote nerve growth factor,suppress oxidative stress and apoptosis,which were pharmacological mechanism to DPN.