| Objective:On the basis of the early study of the relationship between pancreatic microcirculation morphological changes and pancreatic endocrine cell morphology and function expression, Further analyzing the changes of pancreatic microcirculation. Regulation of angiogenic factors as the research object, To find out related material base of the pancreas microcirculation, to find out the related material base of pancreatic microcirculation. From the perspective of angiogenesis to reveal the mechanism of "Ziyinqingre, Yiqihuayu" traditional Chinese medicine compound PRD’s improving T2DM rat’s pancreatic microvascular injury and the improvement of pancreatic microcirculation. And to provide evidence for the curing of T2DM microvascular complications though regulating angiogenic factors and improving pancreatic microcirculation.Methods:1. Animal model replicationMale Wistar rats and the rat are administered intragastrically with fat emulsion everyday for8weeks. Separate serum, examine INS and FPG then inject STZ via tail vein.7days later, examine FPG, and pancreas blood.Then model rats which were successfully replicated were randomized into model group, PRD group (1.77g·kg-1),JiangTangTongMaiPian group (0.42g·kg-1)and Metformin group(0.14g·kg-1) and selected24normal rats as control group.2.The PRD extract preparationThe PRD is composed of six crude drug materials as bellows:lyophilized balsam pear powder(0.6kg), ginseng (1kg), Salvia miltiorrhiza (3kg), Pinellia tuber (1kg), Prepared Radix polygoni multiflori (1kg), Kudzu root(1kg), leech(0.3kg).All crude drugs were purchase from Tong Ren Tang Traditional Chinese Medicine Co., Ltd.(Beijing,China). Momordica charantia L.(Cucurbitaceae) was lyophilized to powder; Panax ginseng C.A. Meyer and Salvia miltiorrhiza Bge were grinded to small granular of approximately5mm and extracted with10times the amount of distilled water three times for2h,1.5h and1h, respectively; Pueraria peduncularis (Grah. ex Benth.) Benth.(Leguminosae) and Polygonum multiflorum Thunb. were extracted with10times the amount of70%ethanol twice for2h and1.5h, respectively, Hirudo medicinalis was extracted with10times the amount of90%ethanol twice for2h and1.5h, respectively. All the above extraction were combined and filtered, and then the filtrate was evaporated under reduced pressure and dried out by spray-drying method. The PRD extract of proper concentration was prepared in distilled water.3. Regulation effects of the Chinese Pu-Ren-Dan Recipe on Ang-1, Ang-2and Tie-2in T2DM Rats with Pancreatic Microcirculation DisturbanceUsing western blot assay to analyze the protein expression changes of Ang-1and Ang-2and phosphorylation levels of Tie-2in each group.RESULTS:PRD reduced T2DM rats’body weight and blood glucose level, resisted the lipid metabolism disturbance, and ameliorated the insulin resistance and β-cell function. In addition, the histological and morphological studies proved that PRD could maintain the normal distribution of endocrine cell in islet and normal ultrastructural of β cell. An increased pancreatic blood flow was observed after PRD treatment. In the investigation of pancreatic angiogenic factors, PRD inhibited the decreased expression of VEGF and Ang-1, and reversed.the reduction of VEGFR2and Tie2phosphorylation in T2DM rats; the Ang-2and TGFβ expression were up-regulated by PRD while PKC was activated; endostain and Angiostatin were down-regulated by PRD.CONCLUSIONS:All these results suggested that increasing VEGF expression, regulating VEGF/VEGFR2signaling, stimulating Ang-1/Tie-2signaling pathway and inhibiting PKC-TGFβ signaling and anti-angiogenic factors might be the underling mechanism of PRD’s anti-diabetic effect, which is contributed to improve the pancreatic vascular system. It could provide a new insight into the prevention and treatment of diabetes. |
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