本文選題：芪芍復(fù)脈湯 + 血管新生��； 參考：《山西省中醫(yī)藥研究院》2017年碩士論文

【摘要】：目的:通過檢測芪芍復(fù)脈湯對下肢缺血大鼠的血清酶學(xué)指標(biāo)、CD34+、缺血骨骼肌毛細(xì)血管密度(MVD)、血管內(nèi)皮細(xì)胞生長因子(VEGF),以及組織病理學(xué)檢測,探討芪芍復(fù)脈湯對大鼠下肢缺血模型血管新生的影響及其可能機(jī)制,為臨床下肢缺血性疾病的治療提供實驗依據(jù)。方法:90只SD大鼠左下肢行股動脈結(jié)扎、分支剔除術(shù),制作下肢缺血模型。隨機(jī)分為模型組,芪芍復(fù)脈湯高、中、低劑量組和粒細(xì)胞集落刺激因子(rhG-CSF)組,每組18只,并設(shè)立假手術(shù)組18只,假手術(shù)組僅作皮膚切開縫合,不結(jié)扎股動脈。術(shù)后第2天開始給藥,芪芍復(fù)脈湯高、中、低劑量組分別按7.6 g·kg-1、3.8 g·kg-1、1.9 g·kg-1灌胃給藥,每天灌胃1次,連續(xù)14天;rhG-CSF組給予rhG-CSF皮下注射0.21ml/100g,每日1次,連續(xù)5天;假手術(shù)組和模型組不給藥,僅予等體積蒸餾水灌胃。分別于術(shù)后3天、7天、14天,每組大鼠分別取6只,用全自動生化儀對大鼠外周血肌酸激酶(CK)、乳酸脫氫酶(LDH)、谷草轉(zhuǎn)氨酶(AST)進(jìn)行檢測;用流式細(xì)胞儀檢測血清中CD34陽性細(xì)胞比例;取術(shù)側(cè)內(nèi)收肌,用免疫組化方法檢測VEGF觀密度值和毛細(xì)血管密度(MVD);用HE病理染色法對術(shù)測腓腸肌病理改變進(jìn)行觀察,比較各組差異。結(jié)果:1.MVD檢測結(jié)果:術(shù)后14天,芪芍復(fù)脈湯高、中劑量組MVD值顯著高于模型組和低劑量組(P0.01);芪芍復(fù)脈湯高劑量組MVD值顯著高于中劑量組(P0.05)。2.酶學(xué)檢測結(jié)果:術(shù)后3天,各組LDH、CK、AST值均較假手術(shù)組顯著升高(P0.05或P0.01);術(shù)后7天,各組(除假手術(shù)組)LDH、CK、AST值均較3天時顯著下降,其中高、中劑量組和rhG-CSF組下降最為明顯(P0.01);術(shù)后14天,各組酶學(xué)LDH、CK、AST指標(biāo)較7天時下降緩慢。3.CD34+陽性細(xì)胞比例檢測結(jié)果:術(shù)后各時間點,芪芍復(fù)脈湯高劑量組CD34+比例均高于模型組(P0.01);術(shù)后7天和14天,芪芍復(fù)脈湯高劑量組CD34+比例明顯高于中劑量組和低劑量組(P0.01)。術(shù)后7天,芪芍復(fù)脈湯高、中劑量組及rhG-CSF組CD34+比例較3天時明顯上升(P0.01),術(shù)后14天,各組(除假手術(shù)組)CD34+比例均下降(P0.05或P0.01)。4.VEGF蛋白表達(dá):術(shù)后各時間點,芪芍復(fù)脈湯高劑量組VEGF觀密度值較模型組顯著上升(P0.01)。術(shù)后7天,除假手術(shù)組外,各組表達(dá)均上升;術(shù)后14天,除假手術(shù)組外,各組表達(dá)均降低。5.病理結(jié)果:芪芍復(fù)脈湯高劑量組肌細(xì)胞萎縮程度較輕,中劑量組次之,高劑量組肌纖維間血管數(shù)目較模型組顯著增多。結(jié)論:芪芍復(fù)脈湯能夠增加缺血骨骼肌毛細(xì)血管密度,促進(jìn)血管新生,其作用機(jī)制可能與上調(diào)VEGF,動員骨髓中的內(nèi)皮祖細(xì)胞向缺血組織遷移、歸巢以促進(jìn)內(nèi)皮細(xì)胞的增生有關(guān)。
[Abstract]:Objective: to detect the serum enzyme index (CD34), capillary density of ischemic skeletal muscle (MVDX), vascular endothelial growth factor (VEGF) and histopathology in rats with lower extremity ischemia treated by Qishaifumai decoction. To explore the effect of Qishaifumai decoction on angiogenesis and its possible mechanism in rat model of lower limb ischemia, and to provide experimental evidence for the treatment of ischemic diseases of lower extremity. Methods 90 Sprague-Dawley rats were treated with femoral artery ligation and branch removal. The model group was randomly divided into three groups: Qishao Fumai decoction group (high, middle and low dose) and granulocyte colony stimulating factor rhG-CSF (rhG-CSF) group (18 rats in each group), and the sham-operation group (18 rats) was used for skin incision and suture without ligation of femoral artery. On the second day after operation, Qishaifumai decoction was given intragastrically at the dose of 7.6g kg ~ (-1) ~ 3.8g ~ (-1) g ~ (-1) ~ (-1) kg-1, respectively. RhG-CSF was given subcutaneously to 0.21ml / 100g, once a day for 5 days for 14 days in the rhG-CSF group. Sham operation group and model group were not given drugs, only equal volume of distilled water intragastric perfusion. Six rats in each group were taken from each group at 3 days and 7 days and 14 days after operation. The levels of creatine kinase (CK), lactate dehydrogenase (LDH) and glutamic oxaloacetic transaminase (AST) in peripheral blood of rats were detected by automatic biochemical instrument, and the percentage of CD34 positive cells in serum was detected by flow cytometry. The adductor muscle was removed and the VEGF and capillary density were detected by immunohistochemical method, and the pathological changes of gastrocnemius muscle were observed by HE pathological staining. Results 1. After 14 days, Qishaifumai decoction was high, the MVD value in the middle dose group was significantly higher than that in the model group and the low dose group, and the MVD value in the high dose group was significantly higher than that in the middle dose group. The results of enzymatic examination: on the 3rd day after operation, the values of LDHK CK AST in each group were significantly higher than those in the sham operation group (P 0.05 or P 0.01), and on the 7th day after operation, the AST values in all groups (except sham-operation group) were significantly lower than those in 3 days after operation, especially in the middle dose group and rhG-CSF group (P 0.01); on the 14th day after operation, there was a significant decrease in the AST value in all groups (except sham-operation group). The results showed that the proportion of CD34 in high dose group of Qishoufumai decoction was higher than that of model group at 7 days after operation, and on the 7th and 14th day after operation, the proportion of CD34 in high dose group was higher than that in model group, and the ratio of CD34 positive cells in each group was higher than that in model group at 7 and 14 days after operation. The proportion of CD34 in high dose group of Qishaifumai decoction was significantly higher than that of medium dose group and low dose group (P 0.01). On the 7th day after operation, Qishaifumai decoction was high, the CD34 ratio in the middle dose group and rhG-CSF group was significantly higher than that in the 3rd day. On the 14th day after operation, the proportion of CD34 in each group (except sham-operation group decreased P0.05 or P0.01).4.VEGF protein expression: at each time point after operation), The apparent density of VEGF in the high dose group of Qishaifumai decoction was significantly higher than that in the model group (P 0.01). On the 7th day after operation, the expression of protein increased in all groups except sham-operation group, and decreased in all groups except sham-operation group on the 14th day after operation. Pathological results: the atrophy degree of muscle cells in the high dose group of Qishaifumai decoction was less than that in the middle dose group, and the number of intermuscular vessels in the high dose group was significantly higher than that in the model group. Conclusion: Qishaifumai decoction can increase capillary density and promote angiogenesis of ischemic skeletal muscle, and its mechanism may be related to up-regulation of VEGF, mobilization of endothelial progenitor cells from bone marrow to migration of ischemic tissue, homing to homing to promote proliferation of endothelial cells.
【學(xué)位授予單位】：山西省中醫(yī)藥研究院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5;R-332

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