复方益糖康对糖尿病大鼠Nav1.7蛋白及mRNA表达的影响

Effect of Chinese Herbal Compound Yitangkang on Nav1.7 Protein and mRNA Expression in Diabetic Rats

目的:探讨中药复方益糖康对链脲佐菌素(STZ)致糖尿病模型大鼠背根神经节(DRG)中电压门控性钠离子通道亚型1.7(Nav1.7)蛋白及mRNA表达的影响。方法:将健康雄性Wistar大鼠40只,体重200～250 g,随机分为空白对照组、模型组和益糖康组,其中,模型组和益糖康组采用STZ 55 mg.kg-1ip复制糖尿病大鼠模型,然后分别给予安慰剂和益糖康(4g.kg-1)5 mL.d-1ig。每周测血糖1次,随时剔除血糖低于16.7 mmol.L-1的大鼠。6周后,采集大鼠背根神经节(DRG)标本,用免疫组化和RT-PCR方法分别检测其钠离子通道Nav1.7蛋白和mRNA表达水平。结果:动物给药6周后处死前血糖:空白对照组(6.73±0.9)mmol.L-1,模型组(20.12±1.3)mmol.L-1,益糖康组(19.34±1.2)mmol.L-1,模型组与空白对照组比较有显著差异(P<0.01),益糖康组与模型组比较无显著差异。Nav1.7蛋白表达的灰度值:空白对照组149.41±5.71,模型组104.53±9.02,益糖康组132.57±6.13,模型组与空白对照组比较其表达水平显著上调(P<0.01),益糖康组与模型组比较其表达水平显著降低(P<0.01)。Nav1.7 mRNA表达的积分吸光度比值:空白对照组0.114±0.018,模型组0.215±0.043,益糖康组0.128±0.025,模型组与空白对照组比较其表达水平显著上调(P<0.01),益糖康组与模型组比较其表达水平显著降低(P<0.01)。结论:中药复方益糖康可能对Nav1.7通道有阻断作用,是其改善糖尿病痛性神经病症状的机制之一。

Objective： To explore the effect of Chinese herbal eompound Yitangkang on Navl. 7 protein and mRNA expression in diabetic rats. Method： Forty health male Wistar rats, weighting 200-250 g, were randomly divided into control group, model group and Yitangkang group. The model group and Yitangkang group were given STZ 55 mg.kg-1 ip to induce diabetes model, and respectively given comfort agent and Yitangkang （4 g.kg-1） 5 mL.d-1 ig for 6 weeks, dorsal root ganglia （DRG） specimens were eollected to detect Navl. 7 protein and mRNA expression level with immunohistochemistry and RT-PCR method respectively. Result： The blood glucose level in control group was （6.73±0.9） mmol.L-1, in model group being （20. 12 ± 1.3） mmol. L-1, in Yitangkang group being （19.34 ±1.2 ） mmol.L-1 , which showed a significant differences in model group compared with the control group （P 〈 0.01 ） , no significant difference in Yitangkang group was found compared with the model group. Grayscale values of the Navl. 7 protein expression in blank control group was 149.41 ± 5.71, in model groups being 104.53 ± 9.02, in Yitangkang group being 132.57 ± 6.13. Compared with the control group, the Navl. 7 protein expression in model group was signifieantly increased （P 〈 0.01 ）, Yitangkang group expression levels significantly lower compared with the model group （P 〈 0.01 ）. Absorbance ratio Nay1.7 mRNA expression of integralin control groups was 0. 114 ± 0. 018, in model groups being 0. 215 ± 0. 043, in Yitangkang group being 0. 128 ± 0. 025. Compared with the control group, the level of expression in model group was significantly increased （ P 〈 0.01 ）. The expression levels in Yitangkang group was significantly lowered compared with the model group （P 〈 0.01 ）. Conclusion ： Chinese herbal compound Yitangkang may be block the Navl. 7 channel, which may be one of the mechanisms.