本文選題：糖尿病性勃起功能障礙　切入點(diǎn)：平滑肌細(xì)胞表型轉(zhuǎn)化　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過(guò)觀察平滑肌細(xì)胞表型轉(zhuǎn)化標(biāo)志蛋白在大鼠陰莖海綿體表達(dá)明確糖尿病勃起功能障礙大鼠(Diabetes-inducederection dysfunction,DI ED)陰莖海綿體是否存在平滑肌細(xì)胞表型轉(zhuǎn)化及表型轉(zhuǎn)化對(duì)勃起的影響;應(yīng)用藥物干預(yù)后,探索五味子醇甲對(duì)DIED大鼠勃起功能的影響及五味子醇甲與平滑肌細(xì)胞表型轉(zhuǎn)化蛋白之間的聯(lián)系。方法:1篩選DIED大鼠:健康雄性SD大鼠100只(體質(zhì)量200±20g),適應(yīng)性喂養(yǎng)1周后,隨機(jī)取88只腹腔注射鏈脲佐菌素(STZ)溶液60mg/kg,余12只為正常對(duì)照組(NC)。72h后測(cè)定隨機(jī)血糖,若血糖16.7mmol/L視為糖尿病(DM)大鼠;繼續(xù)飼養(yǎng)8周,DM大鼠皮下注射阿撲嗎啡(APO),30min內(nèi)未誘導(dǎo)陰莖勃起,視為糖尿病勃起功能障礙(DIED)大鼠;若出現(xiàn)勃起,視為單純DM組大鼠。2模型分組及藥物干預(yù):將DIED大鼠隨機(jī)分為DIED組;五味子醇甲低劑量組(5mg/kg/d);五味子醇甲中劑量組(10mg/kg/d);五味子醇甲高劑量組(20mg/kg/d);另有NC組和DM組共6組;NC組、DM組、DIED組分別予以生理鹽水2ml/d灌胃8周,3個(gè)劑量藥物組予以相應(yīng)劑量五味子醇甲灌胃8周,每日1次。3灌胃8周后麻醉大鼠,電刺激大鼠陰莖海綿體神經(jīng)檢測(cè)陰莖海綿體內(nèi)壓(ICP)、頸動(dòng)脈內(nèi)壓(MAP)。4取血測(cè)定血清谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)、堿性磷酸酶(ALP)、血肌酐(Scr)水平。5斷頭處死,取陰莖海綿體組織,一部分用免疫組化法觀察陰莖海綿體內(nèi)平滑肌肌動(dòng)蛋白-α(SMA-α)水平;另一部分用Western Blot測(cè)定陰莖海綿體內(nèi)SMA-α,骨橋蛋白(OPN)蛋白表達(dá)水平。6數(shù)據(jù)分析:所得數(shù)據(jù)應(yīng)用SPSS 23.0軟件分析,數(shù)據(jù)描述用均數(shù)±標(biāo)準(zhǔn)差((?)±s)表示。對(duì)正態(tài)分布的計(jì)量資料組間比較采用方差分析,多個(gè)樣本均數(shù)間的多重比較采用LSD、SNK-q檢驗(yàn)。以P0.05認(rèn)為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1各組大鼠體質(zhì)量及血糖變化:與NC組相比,灌胃前DM組、DIED組,低、中、高劑量藥物組大鼠體質(zhì)量均明顯下降,血糖顯著增高(P0.05);除NC組外,上述各組間兩兩相比,大鼠體質(zhì)量及血糖無(wú)顯著差異(P0.05)。灌胃8周后,DM組、DIED組,低、中、高劑量藥物組大鼠體質(zhì)量均明顯下降,血糖顯著升高(P0.05),除NC組外,上述各組間兩兩相比,大鼠體質(zhì)量及血糖無(wú)顯著差異(P0.05)。2電刺激前,各組間的基礎(chǔ)ICP無(wú)統(tǒng)計(jì)學(xué)差異(P0.05);電刺激時(shí),各組間MAP值無(wú)統(tǒng)計(jì)學(xué)差異(P0.05);電刺激時(shí),與NC組相比,DM、DIED組大鼠ICP及ICP/MAP依次遞減(P0.05);與DIED組相比,低劑量藥物組ICP及ICP/MAP無(wú)明顯改變(P0.05),中、高劑量藥物組ICP及ICP/MAP依次遞增(P0.05),高劑量藥物組上升幅度最大。3各組大鼠間ALT、AST、ALP及Scr水平均無(wú)明顯差異(P0.05)4免疫組化結(jié)果如圖所示,與NC組比較,SMA-α在DM組、DIED組表達(dá)依次遞減(P0.05);與DIED組比較,SMA-α在低、中、高劑量藥物組表達(dá)依次遞增(P0.05),且不同劑量藥物組SMA-α表達(dá)均低于NC組(P0.05)。5 Western Blot結(jié)果SMA-α蛋白表達(dá):與NC組相比,DM組和DIED組陰莖海綿體中SMA-α蛋白含量減少(P0.05),其中DIED組SMA-α蛋白含量最少(P0.05);與DIED組相比,低、中、高劑量藥物組SMA-α蛋白含量均高于DIED組(P0.05),其中高劑量藥物組SMA-α含量最高,中劑量組次之,再者為低劑量組;不同劑量藥物組蛋白含量均低于NC組(P0.05)。OPN蛋白表達(dá):與NC組相比,DM組和DIED組陰莖海綿體中OPN蛋白含量增多(P0.05),其中DIED組OPN蛋白含量最多(P0.05);與DIED組相比,低、中、高劑量藥物組OPN蛋白含量均低于DIED組(P0.05),但不同劑量藥物組間兩兩比較OPN蛋白表達(dá)無(wú)明顯差異(P0.05);DM組陰莖海綿體中OPN蛋白含量是除NC組外最低者(P0.05)。結(jié)論:1 DIED大鼠陰莖海綿體可能存在平滑肌細(xì)胞表型轉(zhuǎn)化;隨表型轉(zhuǎn)化程度不同可能造成勃起功能障礙。2五味子醇甲干預(yù)后可以改善DIED大鼠勃起功能;五味子醇甲提高勃起功能的作用呈劑量依賴性,高劑量藥物組改善勃起作用最強(qiáng)。3五味子醇甲未對(duì)DIED大鼠肝腎功能相關(guān)指標(biāo)造成影響。4五味子醇甲可能影響了陰莖海綿體中平滑肌細(xì)胞表型轉(zhuǎn)化蛋白的表達(dá),但無(wú)劑量依賴性。
[Abstract]:Objective: to make diabetic erectile dysfunction rat protein in rat corpus cavernosum smooth muscle cell phenotype by observing the signs (Diabetes-inducederection dysfunction, DI ED) whether there is influence on the penis erectile cell phenotype and phenotypic transformation of smooth muscle; application of drug intervention, to explore the effects of schisandrin DIED on erectile function in rats and schisandrin and smooth muscle cell phenotype protein between them. Methods: 1 DIED rats were selected: 100 healthy male SD rats (body weight 200 + 20g), after 1 weeks of feeding, 88 rats were randomly selected by intraperitoneal injection of streptozotocin (STZ) solution 60mg/kg, more than 12 were normal the control group (NC) were determined by random blood glucose.72h, if the blood glucose 16.7mmol/L as diabetes mellitus (DM) rats; to continue for 8 weeks, DM rats by subcutaneous injection of apomorphine (APO), 30min did not induce penile erection, as 涓虹硸灝跨梾鍕冭搗鍔熻兘闅滅(DIED)澶ч紶;鑻ュ嚭鐜板媰璧，

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