当归四逆汤对血瘀证糖尿病周围神经病变大鼠水通道蛋白1活性的影响

Effects of Danggui Sini Decoction on aquaporin1 activity in diabetic peripheral neuropathy rats with blood stasis syndrome

目的:观察当归四逆汤对慢性血瘀证糖尿病周围神经病变(diabetic peripheral neuropathies,DPN)大鼠坐骨神经组织水通道蛋白1(aquaporin1,AQP1)活性的影响,探讨其作用机制。方法:雄性Wistar大鼠40只,分为正常组、模型组、甲钴胺组和当归四逆汤(中药)组,每组10只。除正常组外,均通过喂饲高脂饲料加链脲佐菌素(STZ)腹腔注射造成2型糖尿病模型后,再注射泼尼松龙和肾上腺素制备成2型糖尿病中医血瘀证大鼠模型。当归四逆汤组灌胃给药当归四逆汤(10 g·kg-1·d-1),甲钴胺组灌胃给药甲钴胺(6.75μg·kg-1·d-1),模型组及正常组予等容量0.9%Na Cl溶液,连续8周后处死大鼠取双侧坐骨神经组织。采用干湿重法检测坐骨神经含水量,Western blot方法检测AQP1表达水平。结果:与正常组比较,模型组大鼠活动减少,毛色无光泽,舌质瘀斑紫暗,空腹血糖升高,体质量明显降低(P<0.05);与模型组比较,中药组与甲钴胺组大鼠空腹血糖下降,体质量明显增加(P<0.05)。与正常组比较,模型组大鼠坐骨神经含水量明显升高(P<0.05);与模型组比较,中药组与甲钴胺组大鼠坐骨神经含水量明显下降(P<0.05)。与正常组比较,模型组大鼠AQP1表达水平明显升高(P<0.01);与模型组比较,当归四逆汤组与甲钴胺组大鼠AQP1表达水平明显降低(P<0.05)。结论:血瘀证DPN大鼠AQP1过度表达与坐骨神经组织水停密切相关,当归四逆汤可能通过抑制AQP1的表达干预血瘀水停。

Objective: To observe the effects of Danggui Sini Decoction on the activity of aquaporin1( AQP1) in sciatic nerve tissue of diabetic peripheral neuropathy( DPN) rats with chronic blood stasis syndrome,and discuss its mechanism. Methods: 40 male Wistar rats were divided into the normal group,model group,mecobalamin group and Danggui Sini Decoction group( Chinese medicine group),10 rats in each group. Except the normal group,the model of type 2 diabetes mellitus( T2 DM) was established by high fat diet feeding and streptozotocin( STZ) with intraperitoneal injection,and then the syndrome model of blood stasis was established in the T2 DM rats by prednisolone and epinephrine. The Chinese medicine group was treated with Danggui Sini Decoction at dose of 10 g·kg-1·d-1 by intragastric administration,the mecobalamin group was treated with mecobalamin at dose of 6.75 μg·kg-1·d-1 by intragastric administration,and the model group and normal group were treated with 0.9% Na Cl solution at equivalent volume. After administration for continuous 8 weeks,the rats were sacrificed and the bilateral sciatic nerve tissue were collected. The water content of sciatic nerve was detected by dry/wet weight method. The AQP1 expression level was detected by Western blot. Results: Compared with the normal group,the rats in the model group showed the reduced activity,lusterless hair and dark purple tongue with ecchymosis,and the fasting blood glucose level was increased and the body weight was obviously decreased( P<0.05). Compared with the model group,the fasting blood glucose level was decreased and the body weight was obviously increased in the Chinese medicine group and mecobalamin group( P<0.05). Compared with the normal group,the water content of sciatic nerve was obviously increased in the model group( P<0.05). Compared with the model group,the water content of sciatic nerve was obviously decreased in the Chinese medicine group and mecobalamin group( P<0.05). Compared with the normal group,the AQP1 expression level was obviously increased in the model group( P < 0.01). Compared with the model group,the AQP1 expression level was obviously decreased in the Chinese medicine group and mecobalamin group( P<0.05). Conclusion: The overexpression of AQP1 is closely related to the water content of sciatic nerve tissue in DPN rats with blood stasis syndrome. Danggui Sini Decoction may inhibit the AQP1 expression and improve the blood stasis and fluid retention.