托里消毒散精简方对糖尿病皮肤溃疡大鼠创面愈合的影响及其机制

Effects of Tuoli Xiaodu Powder on healing of wound in diabetic rats with skin ulcer and its mechanism

目的探讨托里消毒散精简方全方及其拆方影响糖尿病皮肤溃疡大鼠创面愈合的作用机制。方法 200只雄性SD大鼠随机分为对照组、模型组、托里消毒散精简方全方(黄芪、当归、白芷、皂角刺)组、托法(黄芪、当归)组、透法(白芷、皂角刺)组,采用尾iv链脲佐菌素(STZ)50 mg/kg制备糖尿病模型,同时背部打孔法制备糖尿病皮肤溃疡大鼠模型。各组分别ig给予托里消毒散精简方全方及其拆方的浸膏,对照组和模型组ig等量生理盐水,连续给药14 d,分别于创伤后5、8、11、14 d评价创面溃疡愈合情况。采用HE染色观察创面肉芽组织形态学改变;免疫组织化学法检测创面组织血管生成素受体2(Tie-2)、血管生成素1(Ang-1)、血管生成素2(Ang-2)在肉芽组织中的定位表达;Western blotting法检测创面肉芽组织中Tie-2、Ang-1、Ang-2的蛋白表达水平。结果创伤后5、8、11、14 d,各药物组大鼠创面愈合率均高于模型组(P<0.05);创伤后5 d,透法组愈合率高于托法组和全方组(P<0.05);创伤后8、11、14 d,全方组愈合率高于托法组和透法组(P<0.05);全方组各时间点肉芽组织中Ang-1、Tie-2蛋白表达水平均高于模型组(P<0.05),Ang-2蛋白表达水平均低于模型组(P<0.05);创伤后5、8 d,透法组Ang-2蛋白表达水平明显下调(P<0.05);创伤后11、14 d,托法组Ang-1、Tie-2蛋白表达水平上调(P<0.05)。结论托里消毒散精简方全方可能通过上调创面肉芽组织中Ang-1、Tie-2的蛋白表达水平,下调Ang-2的蛋白表达水平促进创面愈合。

Objective To study the possible mechanism of Tuoli Xiaodu Powder and disassembled prescriptions for the healing of wound in diabetic rats with skin ulcer.Methods Totally 200 male SD rats were randomly divided into control group,model group,tuo disassembled prescription group,tou disassembled prescription group,and whole prescription group.The diabetes rat model was established using caudal vein injection of Streptozotocin(STZ) and the skin ulcer model was prepared by 'back drilling' method.The rats in each group were given different prescriptions and corresponding physiological saline.The healing conditions of ulcer were observed on days 5,8,11,and 14 after treatment.The structural changes of granulation tissue were observed by HE staining.Expression of Tie-2,Ang-1,and Ang-2 in the granulation tissues of the wound were detected by Western blotting and immunohistochemistry.Results The healing rate of each treatment group was higher than that of the DM group at all the time(P < 0.05); The healing rate of the tou group was higher than that of the tuo group and whole group on day 5 after treatment(P < 0.05); The healing rate of the whole group was higher than that of the tuo and tou group(P < 0.05).The expression of Ang-1 and Tie-2 was increased and that of Ang-2 was decreased in the whole group compared with the DM group(P < 0.05).Expression of Ang-2 in the tou group was significantly decreased on days 5 and 8 after treatment(P < 0.05).Expression of Ang-1 and Tie-2 in the tuo group was increased on days 11 and 14 after treatment(P < 0.05).Conclusion The increased expression of Ang-1 and Tie-2 in the granulation tissue and the decreased expression of Ang-2 maybe the mechanism through which the Tuoli Xiaodu Powder promoting wound healing.