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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 therapeutic effect of Jiangtangtongluo decoction on peripheral neuropathy (DPN) in experimental diabetic rats and its mechanism of action by promoting NGF, anti-oxidation and anti-apoptosis. Methods: DPN model was established by intravenous injection of streptozotocin (60mg / kg). The rats were randomly divided into model control group, shangxiao tongluo decoction 36,18,9g crude drug / kg group, metformin hydrochloride mecobalamin dispersible tablets Group (cell experiment Jinqijing hypoglycemic tablets) for 8 weeks, 1 time / d. The changes of fasting blood glucose and photothermal radiation pain threshold were observed 2 and 8 weeks after administration. The expression of nerve growth factor (NGF) protein in sciatic nerve was detected by immunohistochemistry. The expression of NGF mRNA was detected by RT-PCR. IF was used to measure the expression of inducible nitric oxide synthase (iNOS) in high glucose-cultured SC cells and the expression of p22-phox subunit of NADPH oxidase in high glucose-cultured Schwann cells by RT-PCR. The expression of Casepase-3 mRNA and Bcl-2 mRNA was detected by RT-PCR and the apoptosis of Schwann cells was detected by TUNEL method. Results: Jiangtong Tongluo Decoction 36,18 g crude drug / kg group can significantly reduce fasting blood glucose (P <0.01), shorten the incubation period of heat source stimulation (P <0.01). (P <0.01), inhibit the expressions of iNOS and p22-phox m RNA in NADPH (P <0.01), and inhibit the expression of Casepase-3 mRNA in the sciatic nerve (P <0.01) (P <0.01), inhibit the apoptosis rate. Conclusion: Jiangtang Tongluo Decoction can improve peripheral nerve lesion through promoting NGF, inhibiting oxidative stress and controlling apoptosis.