| Objective1. To investigate the effect of Simiaoyongan recipe on angiogenesis for diabetic ratsof hindlimb ischemia;2. To observe the impact of Simiaoyongan recipe on the protein of Akt andp38MAPK in PI3K/Akt and p38MAPK signal pathways;3. To establish the hindlimb ischemic model on Wistar rat with diabetes mellitus.Methods1. Diabetic rat models were induced with Streptozotocin, and a hindlimb ischemicmodels were prepared in40Wister rats by ligation and excision of left femoralartery and its branches. All rats were randomly divided into4groups: controlgroup, control therapeutic group, diabetic group, diabetic therapeutic group.Simiaoyongan recipe were given intragastric administration on controltherapeutic group and diabetic therapeutic group, while distilled water were givenon control group and diabetic group.2. Blood flow(Perfusion,PU) were performed with doppler scan before and afteroperation, and at1,3,7,14days after operation. The pathological changes of newvascular were observed by CD31immunohistochemistry and the expressions ofAkt and p38MAPK were measured by Western-Blot on15days after operation.Results1. Control group, control therapeutic group, diabetic group and diabetic therapeutic group blood glucose is(mmol/L)[Day3(6.24±0.45),(6.10±0.37),(24.87±4.47),(26.09±3.95);Day7:(6.60±0.87),(6.17±0.39),(26.77±3.60),(28.10±4.15)].In contr-ast to control and control therapeutic group, blood glucose of diabetic group anddiabetic therapeutic group are significantly increased at three days afterintragastric administration of streptozotocin, which can reach a plateau at7daysafter intragastric administration and maintain more than21days. And we took anoperation for hindlimb ischemic model, which is obviously effective. Comparedwith preoperation PU ratio is a marked reduction after operation in control group,control therapeutic group, diabetic group and diabetic therapeutic group, butthere is no remarkable difference between the four groups.2. Control group, control therapeutic group and diabetic group PU ratio is(%)[Day7:(45±4),(49±3),(36±3);Day14:(63±6),(66±4),(45±4)]. Compared with controland control therapeutic group, the PU ratio and the number of new vascular ofdiabetic group were significantly decreased, as well as angiogenesis ability defect,and the expression of p-Akt were obviously decreased, but p38MAPK wasobviously increased(P<0.05).3. While the improvement of angiogenesis was obvious on diabetic therapeuticgroup, the PU ratio [diabetic group(%): Day7:(36±3), Day14:(45±4); diabetictherapeutic group: Day7:(48±6), Day14:(61±4),P<0.05],the number of newvascular of were significantly higher than diabetic group[diabetic group:(9.1±2.0),diabetic therapeutic group:(16.4±2.9),P<0.05], as well as ratio (operated side vsun-operated side)[diabetic group:(0.62±0.15), diabetic therapeutic group:(0.86±0.18),P<0.05],the expression of p-Akt were obviously increased[diabetic therap-eutic group:(0.28±0.56) vs diabetic group:(0.20±0.07),P<0.05],and p38MAPKwas obviously decreased[diabetic therapeutic group:(0.99±0.04) vs diabeticgroup:(1.18±0.23),P<0.05]. ConclusionWe successfully established an diabetic rat model and hindlimb ischemic model,the method can lead rats more drink, more urine, and high blood glucose which istypical characteristic for type2diabetes, we took operation to make hind limbsischemic model, ischemic effect is obvious. It is a effective, convenient methodto establish a diabetic rats of limbs ischemic model combined with both methodsabove. Simiaoyongan recipe could significantly improve and accelerateangiogenesis process on diabetic rats of limb ischemia, which possible relates toup-reguation of Akt signal pathway, and down-regulation of p38MAPK signalpathway. |
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