Study Of Pilose Antler Promote Angiogenesis In Rats Hind Limb Ischemia Model By Mobilizing EPCs

ObjectiveTo observe the impacts of pilose antler on the number and the proliferative ability of EPCs, the contents of NO and the expression of protain VEGF in the rats hind limb ischemia models so as to explore the mechanism of pilose antler promotes the treatments of angiogenesis by mobilizing the EPCs. MethodThe model were established the left leg branch of femoral artery ligation and stripping out, due to hind limb vascular ischemia. The modeled rats were randomized into a model group, pilose antler high, middle, low dose groups, compound salvia table group. Among the normal rats of same batch, only not ligation femoral artery and its branches, were selected randomly to set up a normal control group. 15 rats in each group.The gavage was given once a day in the rats of each group two days later after the operation, in accordance with the 0.572g·kg-1, 0.286 g·kg-1, 0.143 g·kg-1for postoperative lavage. The dosage of compound salvia table group is 0.286g·kg-1. The stilled water of same dose was used in the normal control group and the model group. 5 rats were selected randomly from each group respectively on the 3rd day, the 7th day, the 14 th day after operation groups. Automatic biochemical analyzer was used to detect the expression level of aspartate aminotransferase(AST), creatine kinase(CK), lactate dehydrogenase(LDH) in the rat serum. Left thigh gastrocnemius muscle were analyzed by immunohistochemistry method. To observe the recovery of the capillaries in the ischemic limb, and its capillary density was calculated. Hematoxylin eosin(he) staining was used to observe the pathological change of the left gastrocnemius muscle. One week later, the isolation and culture of EPCs were performed. Flow cytometry instrument was used to identify CD34/CD133 phenotype. Cell counting kit-8 was applied to check the number and proliferation ability of EPCs. Protain VEGF was checked by ELISA kit, and the content of NO was measured by nitric acid deoxidize enzyme method. Results1. The positive expression of CD34/CD133 was identified by flow cytometry, which showed that endothelial progenitor cells were found in each group.2. Pilose antler could promote the the number and proliferation ability of EPCs, so as to increase the content of NO, and it can also enhance the expression of VEGF.3. Pilose antler high-dose group were able to significantly lower the levels of AST, CK and LDH in limb ischemia rats serum on the 3rd day, the 7th day, the 14 th day after operation, which coincide with the previous experimental results.4. Pilose antler high dose group significantly improved the value of MVD in the rats with hind limb ischemia on the 7th day and the 14 th day after operation, but on the third day after operation, the effect is not obvious.5. Pilose antler can improve muscle cell atrophy in the rats with hind limb ischemia, alleviate interstitial hyperplasia pathological changes and increase muscle fiber between vessels. ConclusionPilose antler could mobilize endothelial progenitor cells to repair endothelial function, promote angiogenesis, and abate limb ischemia condition. In order to provide some basis for the treatment of lower limb ischemia diseases, which has a far-reaching significance.