本文關(guān)鍵詞：阿托伐他汀對糖尿病下肢動脈硬化PTA術(shù)后再狹窄影響的實驗研究　出處：《山東大學(xué)》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 阿托伐他汀 再狹窄 經(jīng)皮腔內(nèi)血管成形術(shù) 下肢血管病變 糖尿病 阿托伐他汀 再狹窄 經(jīng)皮腔內(nèi)血管成形術(shù) 下肢血管病變 糖尿病

【摘要】：目的糖尿病患者動脈粥樣硬化(atherosclerosis,AS)的患病率較高,而且容易發(fā)生在主動脈,冠狀動脈,肢體外周動脈等部位。冠狀動脈目前多采用血管內(nèi)支架植入術(shù),但是肢體外周AS常以下肢動脈病變?yōu)橹?尤其是膝以下動脈及其分支。由于內(nèi)徑細(xì)、壓力低,膝下動脈的治療一直是個難題。新型的球囊導(dǎo)管由于剖面低、壓力低、長段、順應(yīng)性較好的優(yōu)點,大大增加了膝下動脈進(jìn)行介入治療的可行性。因此經(jīng)皮腔內(nèi)血管成形術(shù)(percutaneous transluminal angioplasty,PTA)成為目前治療糖尿病下肢動脈特別是膝下AS的重要手段。但由于PTA術(shù)后再狹窄發(fā)生率高,平均約為30%,嚴(yán)重影響了 PTA的遠(yuǎn)期效果。如何防治PTA術(shù)后再狹窄成為影響糖尿病下肢AS治療的關(guān)鍵問題。他汀類藥物,是羥甲基戊二酰輔酶A(hydroxy-methylglutaryl-CoA,HMG-CoA)還原酶抑制劑。實驗研究表明,除了降低膽固醇的作用外,它們還有非調(diào)脂的抗AS作用:抑制血管內(nèi)皮的炎癥反應(yīng),穩(wěn)定粥樣斑塊,改善血管內(nèi)皮功能,抑制動脈內(nèi)膜增生等�；谏鲜鲈�,他汀類藥物在臨床上用于AS患者PTA術(shù)后再狹窄的預(yù)防。但是PTA術(shù)后再狹窄與AS的產(chǎn)生機制不盡相同,血管平滑肌細(xì)胞(vascular smooth muscle cells,VSMCs)的增殖與遷移在再狹窄的進(jìn)程中起了非常關(guān)鍵的作用。盡管部分體外實驗觀察到他汀類藥物能夠抑制VSMCs的增殖、遷移、凋亡,但是有臨床試驗研究發(fā)現(xiàn)他汀類藥物抑制再狹窄的臨床療效欠佳。他汀類藥物是否可以抑制再狹窄?特別是糖尿病下肢AS的PTA術(shù)后再狹窄?這是尚未明確的問題。臨床獲得血管標(biāo)本具有局限性,使得我們需要動物體內(nèi)實驗提供更直觀和確切的證據(jù)。目前很多研究采用單次球囊拉傷術(shù)建立再狹窄動物模型。這種模型的缺點在于:直接在正常的血管上而不是發(fā)生AS的血管上建立再狹窄模型,與人體血管內(nèi)再狹窄發(fā)生的過程有較大差異。為了明確上述問題,我們的研究目的總結(jié)如下:首先,我們的實驗先通過一次球囊拉傷術(shù)建立糖尿病兔髂動脈AS模型,然后在此基礎(chǔ)上進(jìn)行PTA手術(shù),更好的模擬了糖尿病患者下肢血管硬化PTA術(shù)后再狹窄的過程,應(yīng)用雙次球囊拉傷術(shù)/擴張術(shù)為再狹窄的研究建立更為準(zhǔn)確的動物模型。其次,通過觀察PTA術(shù)后不同時間點的狹窄率、內(nèi)膜增生情況、VSMCs在血管內(nèi)的分布情況等以明確PTA術(shù)后再狹窄的進(jìn)程和特點。第三,通過應(yīng)用他汀類藥物,觀察其對血管平滑肌細(xì)胞的遷移、增殖以及再狹窄程度的影響以明確其是否可以抑制PTA術(shù)后再狹窄的發(fā)生。方法一、建模與分組選取雄性新西蘭白兔(體重約1.7kg),適應(yīng)性喂養(yǎng)1周后,給予高脂飼料喂養(yǎng)至實驗終止。1、雙次球囊拉傷術(shù)/擴張術(shù)再狹窄模型建立(1)糖尿病模型建立:耳緣靜脈注射四氧嘧啶(劑量為80mg/kg),1周后測量血糖確認(rèn)糖尿病模型的建立。(2)AS模型建立:糖尿病模型建立后1周對髂動脈行球囊拉傷術(shù),拉傷術(shù)后4周通過超聲檢查確認(rèn)髂動脈AS模型的建立。(3)再狹窄模型建立:在AS形成的血管部位行PTA手術(shù)以建立再狹窄模型。2、實驗分組與處理將造模成功的再狹窄模型動物隨機分為四組:7天再狹窄組,14天再狹窄組,28天再狹窄組,28天阿托伐他汀組,另設(shè)對照組。各組處理方法(1)7天再狹窄組:給予相同體積生理鹽水灌胃,PTA術(shù)后第7天處死。(2)14天再狹窄組:給予相同體積生理鹽水灌胃,PTA術(shù)后第14天處死(3)28天再狹窄組:給予相同體積生理鹽水灌胃,PTA術(shù)后第28天處死。(4)28天阿托伐他汀組:給予常規(guī)劑量阿托伐他汀(2.5mg/kg/d)灌胃28天后處死。(5)對照組:進(jìn)行和再狹窄模型組動物相同的外科手術(shù),但不進(jìn)行球囊擴張,相同體積生理鹽水灌胃。二、標(biāo)本留取及指標(biāo)檢測1、根據(jù)實驗分組,分別在不同時間點(PTA術(shù)后第7天,14天,28天)處死動物,留取血液樣本及髂動脈標(biāo)本。2、血脂指標(biāo)的檢測:(1)血清總膽固醇(total cholesterol,TC);(2)甘油三酯(Triglyceride,TG);(3)低密度脂蛋白(low density lipoprotein cholesterin,LDL-C);(4)高密度脂蛋白(high density lipoprotein cholesterol,HDL-C)。3、血管形態(tài)學(xué)檢測:髂動脈段固定在4%多聚甲醛中,脫水后包埋于石蠟中切片,進(jìn)行以下檢測(1)通過HE染色觀察血管的一般形態(tài)(2)通過Masson染色觀察血管平滑肌的形態(tài)及膠原蛋白(3)通過EVG染色觀察彈力纖維形態(tài)。(4)應(yīng)用Image-Pro Plus 6.0圖像分析軟件測量血管斑塊位置的血管腔面積,內(nèi)膜、中膜面積等指標(biāo)以計算血管狹窄率和內(nèi)膜/中膜比值。4、免疫組化分析:通過增殖細(xì)胞核抗原(proliferating cell nuclear antigen,PCNA)的免疫組化分析測定血管再狹窄位置內(nèi)膜和中膜增殖細(xì)胞的數(shù)目和去分化情況。PCNA指數(shù)根據(jù)以下公式計算:PCNA陽性細(xì)胞數(shù)/總細(xì)胞計數(shù)×100%5、免疫熒光染色:通過αa-平滑肌肌動蛋白(alpha smooth muscle actin,α-SMA)的免疫熒光染色觀察VSMCs從中膜向內(nèi)膜遷移和分化的情況。二、統(tǒng)計學(xué)分析所有的數(shù)據(jù)應(yīng)用SPSS20.0統(tǒng)計軟件進(jìn)行處理。組間的比較采用了單因素方差分析,所有的數(shù)據(jù)均以均數(shù)±標(biāo)準(zhǔn)差(x±s)的方式表示,P0.05被認(rèn)為組間差異有統(tǒng)計學(xué)意義。結(jié)果一、動物成模及血糖情況45只新西蘭白兔在糖尿病模型建立過程中,5只死亡,10只血糖水平未達(dá)標(biāo),AS及再狹窄模型建立過程中全部成功。各組實驗動物的血糖在整個實驗過程中始終保持在300mg/dl(16.7mmol/L)的水平之上。二、血脂指標(biāo)檢測結(jié)果.與28天再狹窄組以及對照組相比,28天阿托伐他汀組的TG、TC和LDL-C水平出現(xiàn)下降(P0.05)。而未用藥的各再狹窄組(7天再狹窄組、14天再狹窄組、28天再狹窄組)與對照組之間TG、TC和LDL-C水平未見顯著統(tǒng)計學(xué)差異(P0.05)。上述所有實驗組之間的HDL-C水平均未見顯著統(tǒng)計學(xué)差異(P0.05)。三、不同時間點再狹窄進(jìn)展情況1、血管形態(tài)學(xué)檢測:HE染色:與對照組相比,各再狹窄組管腔明顯變窄,內(nèi)膜增厚。Masson染色:增厚的內(nèi)膜主要是平滑肌細(xì)胞及基質(zhì)。EVG染色:增厚的內(nèi)膜主要是彈力纖維,內(nèi)彈力板出現(xiàn)斷裂,中膜比例縮小以及中膜彈力纖維斷裂。隨著時間的推移,從PTA術(shù)后第7天到第28天,上述病理改變更加典型,程度逐漸加深。2、血管狹窄率和內(nèi)膜/中膜比值計算顯示:與對照組相比,各再狹窄組的狹窄率明顯增加,差別具有統(tǒng)計學(xué)意義(P0.05)。各再狹窄組的狹窄率,從第7天到14天到28天逐漸增加,差別具有統(tǒng)計學(xué)意義(P0.05),而且從第7天到第14天增加速度更顯著。各再狹窄組的內(nèi)膜厚度以及內(nèi)膜/中膜比的增加趨勢與狹窄率一致,差別同樣具有統(tǒng)計學(xué)意義(P0.05)。結(jié)果表明再狹窄的發(fā)生是逐漸進(jìn)展,而且最初的進(jìn)展速度更快。3、α-SMA的免疫熒光染色顯示:對照組的動脈中膜中a-SMA強陽性表達(dá)而各再狹窄組的動脈中膜中α-SMA的陽性表達(dá)不連續(xù)。對照組動脈內(nèi)膜中未見a-SMA陽性細(xì)胞,各再狹窄組的動脈內(nèi)膜中可見a-SMA陽性細(xì)胞。從PTA術(shù)后第7天、第14天到第28天,α-SMA陽性細(xì)胞逐漸增多,VSMCs的增加趨勢與狹窄率一致。4、PCNA免疫組化分析顯示:與上述指標(biāo)不同,PCNA指數(shù)在7天再狹窄組最大。14天再狹窄組與7天再狹窄組相比PCNA指數(shù)出現(xiàn)下降,14天再狹窄組與28天再狹窄組相比未見顯著統(tǒng)計學(xué)差異(P0.05)。表明內(nèi)膜細(xì)胞增殖在第7天得到最大程度的誘導(dǎo),隨后下降并穩(wěn)定在一定水平。雖然繼續(xù)保持增殖的趨勢,但增殖的速度在第7天后有所下降并逐漸保持穩(wěn)定。四、常規(guī)劑量阿托伐他汀對再狹窄的影響1、血管狹窄率和內(nèi)膜/中膜比值計算顯示:與28天再狹窄組相比,28天阿托伐他汀組的狹窄率和內(nèi)膜/中膜比沒有明顯的下降,未見顯著統(tǒng)計差異(P0.05)。常規(guī)劑量的阿托伐他汀治療沒有明顯抑制再狹窄的發(fā)生。2、α-SMA的免疫熒光染色顯示:與28天再狹窄組相比,28天阿托伐他汀組的內(nèi)膜中仍然觀察到一定數(shù)量的VSMCs,兩組之間未見明顯差別。3、PCNA免疫組化分析顯示:與28天再狹窄組相比,28天阿托伐他汀組的PCNA指數(shù)未見顯著統(tǒng)計學(xué)差異(P0.05)。上述結(jié)果顯示:常規(guī)劑量的阿托伐他汀在實驗動物體內(nèi)沒有明顯抑制VSMCs向內(nèi)膜的遷移和增殖,進(jìn)一步驗證了常規(guī)劑量阿托伐他汀的治療沒有明顯抑制PTA術(shù)后再狹窄的發(fā)生。結(jié)論一、本研究通過球囊拉傷術(shù)先建立糖尿病動物AS模型,然后在AS基礎(chǔ)上進(jìn)行PTA手術(shù),進(jìn)而建立再狹窄動物模型,更好的模擬了糖尿病下肢AS患者PTA術(shù)后再狹窄的發(fā)生過程,通過雙次球囊拉傷術(shù)/擴張術(shù)成功建立了理想的PTA術(shù)后再狹窄動物模型。二、在動物模型中,PTA術(shù)后再狹窄的發(fā)生發(fā)展呈現(xiàn)時間相關(guān)性。從第7天到第14天,再狹窄程度進(jìn)展迅速;從第14天到第28天,再狹窄的進(jìn)展速度有所減緩,但狹窄程度已經(jīng)非常嚴(yán)重。內(nèi)膜中VSMCs的增殖在第7天得到最大程度的誘導(dǎo),隨后增殖速度有所減緩,但第28天仍有一定程度的增殖。三、他汀類藥物雖然能夠防治AS,而且部分體外和體內(nèi)實驗觀察到他汀類藥物可以抑制VSMCs增殖、遷移,但是在我們的實驗動物體內(nèi),常規(guī)劑量的阿托伐他汀對VSMCs向內(nèi)膜的遷移和增殖以及再狹窄的發(fā)生發(fā)展無顯著影響。目的:我們第一部分的實驗研究結(jié)果表明:常規(guī)劑量的阿托伐他汀對VSMCs在動物模型體內(nèi)的遷移、增殖以及再狹窄的發(fā)生發(fā)展均無顯著影響。在第一部分實驗的討論中,我們對出現(xiàn)陰性結(jié)果的可能原因進(jìn)行了總結(jié)。其中血藥濃度不足可能是導(dǎo)致陰性結(jié)果的重要原因之一。血藥濃度不足分為絕對不足和相對不足。血藥濃度絕對不足是由于他汀類藥物作用的劑量依賴性。劑量依賴性是指可根據(jù)藥物劑量調(diào)整在一定范圍內(nèi)提高療效。有研究結(jié)果表明,他汀類藥物對VSMCs的增殖、遷移,抑制冠狀動脈PCI術(shù)后再狹窄,抑制再狹窄的部位炎癥反應(yīng),保護(hù)損傷內(nèi)皮等作用均呈劑量依賴性。血藥濃度相對不足與不同動物模型相關(guān)。很多再狹窄的研究采用單次球囊拉傷模型,但這種模型是損傷正常血管造成的再狹窄,程度較輕,不夠貼近人體內(nèi)再狹窄的過程。而我們的雙次損傷模型是在已經(jīng)發(fā)生AS的血管上造成的再狹窄,程度較重,更貼近人體內(nèi)再狹窄的過程。血藥濃度可能因為嚴(yán)重的狹窄而相對不足,因此需要進(jìn)行強化劑量的阿托伐他汀治療才有可能獲得理想的效果。論文第二部分的研究目的可以概括為:通過應(yīng)用不同劑量的阿托伐他汀,觀察強化劑量阿托伐他汀是否會對血管平滑肌細(xì)胞的遷移、增殖以及再狹窄程度產(chǎn)生與常規(guī)劑量阿托伐他汀不一樣的效果,阿托伐他汀對再狹窄的影響是否呈現(xiàn)劑量依賴性。方法一、建模與分組選取雄性新西蘭白兔(體重約1.7kg),適應(yīng)性喂養(yǎng)1周后,給予高脂飼料喂養(yǎng)至實驗終止。1、再狹窄模型建立(1)糖尿病模型建立:耳緣靜脈注射四氧嘧啶(劑量為80mg/kg),1周后測量血糖確認(rèn)糖尿病模型的建立。(2)AS模型建立:糖尿病模型建立后1周對髂動脈行球囊拉傷術(shù),拉傷術(shù)后4周通過超聲檢查確認(rèn)髂動脈AS模型的建立。(3)再狹窄模型建立:在AS形成的血管部位行PTA手術(shù)以建立再狹窄模型。2、實驗分組與處理將造模成功的再狹窄模型動物隨機分為三組:(1)空白對照組(簡稱對照組)PTA手術(shù)當(dāng)天開始給予與常規(guī)組相同體積的生理鹽水灌胃至PTA術(shù)后第28天。(2)常規(guī)劑量阿托伐他汀組(簡稱常規(guī)組):PTA手術(shù)當(dāng)天開始給予常規(guī)劑量阿托伐他汀(2.5mg/kg/d)灌胃至PTA術(shù)后第28天。(3)強化劑量阿托伐他汀組(簡稱強化組):PTA手術(shù)當(dāng)天開始給予強化劑量阿托伐他汀(l0mg/kg/d)灌胃至PTA術(shù)后第28天。二、標(biāo)本留取及指標(biāo)檢測1、PTA術(shù)后第28天處死動物,留取血液樣本及髂動脈標(biāo)本。2、血液指標(biāo)檢測:TC,TG,LDL-C和HDL-C。3、血管形態(tài)學(xué)檢測:髂動脈段固定在4%多聚甲醛中,脫水后包埋于石蠟中切片,進(jìn)行以下檢測(1)通過Masson染色觀察血管平滑肌的形態(tài)及膠原蛋白(2)通過EVG染色觀察彈力纖維形態(tài)。(3)應(yīng)用Image-Pro Plus 6.0圖像分析軟件測量血管斑塊位置的血管腔面積,內(nèi)膜、中膜面積等指標(biāo)以計算血管狹窄率和內(nèi)膜/中膜比值。4、PCNA免疫組化分析:測定血管再狹窄位置內(nèi)膜和中膜增殖細(xì)胞的數(shù)目和去分化情況。PCNA指數(shù)根據(jù)以下公式計算:PCNA陽性細(xì)胞數(shù)/總細(xì)胞計數(shù)×100%5、a-SMA免疫組化染色:觀察VSMCs從中膜向內(nèi)膜遷移和分化的情況。三、統(tǒng)計學(xué)分析所有的數(shù)據(jù)應(yīng)用SPSS 20.0統(tǒng)計軟件進(jìn)行處理。組間的比較采用了單因素方差分析,所有的數(shù)據(jù)均以均數(shù)土標(biāo)準(zhǔn)差(x±s)的方式表示,P0.05被認(rèn)為組間差異有統(tǒng)計學(xué)意義。結(jié)果一、動物成模及血糖情況30只新西蘭白兔在糖尿病模型建立過程中:3只死亡,9只血糖水平未達(dá)標(biāo),18只成為糖尿病兔。18只糖尿病兔建立再狹窄模型過程中有3只死亡,15只造模成功。各組實驗動物的血糖在整個實驗過程中始終保持在300mg/dl(16.7mmol/L)的水平之上。二、血脂檢測結(jié)果與對照組相比,常規(guī)組和強化組的TG、TC和LDL-C水平出現(xiàn)下降(P0.05)。常規(guī)組與對照組之間的HDL-C水平未見顯著統(tǒng)計學(xué)差異(P0.05),與前兩組相比,強化組HDL-C水平升高,有統(tǒng)計學(xué)差異(P0.05)。三、不同劑量阿托伐他汀對再狹窄的影響1、血管狹窄率和內(nèi)膜/中膜比值計算顯示:與對照組相比,常規(guī)組和強化組的狹窄率和內(nèi)膜/中膜比均無明顯下降,未見顯著統(tǒng)計差異(P0.05)。常規(guī)劑量和強化劑量阿托伐他汀的治療都沒能明顯抑制再狹窄的發(fā)生。2、α-SMA的免疫組化染色顯示:與對照組相比,常規(guī)組和強化組內(nèi)膜中的α-SMA陽性細(xì)胞均未見明顯減少的趨勢。常規(guī)劑量和強化劑量阿托伐他汀的治療均沒能明顯減少內(nèi)膜中分化型VSMCs的數(shù)量,即沒能抑制抑制VSMCs向內(nèi)膜的遷移與增殖。3、PCNA免疫組化分析顯示:與對照組相比,常規(guī)組和強化組的PCNA指數(shù)均無明顯下降,未見顯著統(tǒng)計學(xué)差異(P0.05)。常規(guī)劑量和強化劑量阿托伐他汀的治療沒能明顯降低再狹窄部位增殖細(xì)胞的數(shù)量,即使強化治療也沒能抑制再狹窄部位細(xì)胞的增殖與去分化。結(jié)論強化劑量的阿托伐他汀與常規(guī)劑量的阿托伐他汀相似,對VSMCs在體內(nèi)的遷移、增殖、去分化以及再狹窄發(fā)生發(fā)展的影響均無顯著效果。在我們的糖尿病下肢動脈硬化PTA術(shù)后再狹窄的模型中,阿托伐他汀對再狹窄的影響未呈現(xiàn)劑量依賴性。
[Abstract]:Objective the prevalence of atherosclerosis (atherosclerosis, AS) is high in diabetic patients, and is prone to occur in aorta, coronary artery, peripheral artery and other parts. The coronary artery stent implantation is often used at present, but the peripheral AS of the extremities is often dominated by lower extremity artery disease, especially the subgenu artery and its branches. The treatment of the inferior genicular artery has been a difficult problem because of the low internal diameter and low pressure. The new balloon catheter has the advantages of low profile, low pressure, long length, and good compliance, which greatly increases the feasibility of the interventional treatment of the inferior genicular artery. Therefore, percutaneous transluminal angioplasty (percutaneous transluminal angioplasty, PTA) has become an important way to treat diabetic lower extremity artery, especially below knee AS. However, the incidence of restenosis after PTA is high, with an average of about 30%, which seriously affects the long-term effect of PTA. How to prevent and control restenosis after PTA is the key problem that affects the AS treatment of diabetic lower extremity. The statins are the hydroxyl methylamyl two acyl coenzyme A (hydroxy-methylglutaryl-CoA, HMG-CoA) reductase inhibitor. Experimental studies show that besides lowering cholesterol, they also have non lipid lowering anti AS effects: inhibiting inflammatory reaction of vascular endothelium, stabilizing atherosclerotic plaques, improving vascular endothelial function and inhibiting intimal hyperplasia. For the above reasons, statins are clinically used for the prevention of restenosis after PTA in AS patients. However, the mechanism of restenosis after PTA is different from that of AS. The proliferation and migration of vascular smooth muscle cells (VSMCs) play a key role in the process of restenosis. Although some statins in vitro inhibit the proliferation, migration and apoptosis of VSMCs, clinical trials have found that statins inhibit the restenosis. Does statins inhibit restenosis, especially in diabetic lower extremity AS after PTA restenosis? This is a problem that is not yet clear. The limitations of clinical acquisition of vascular specimens make it necessary for us to provide more intuitive and accurate evidence in animal experiments. In many studies, a single balloon injury is used to establish an animal model of restenosis. The disadvantage of this model is that the establishment of restenosis models directly on normal blood vessels rather than AS vessels is quite different from that in human vascular restenosis. In order to solve this problem, the aim of our study is summarized as follows: firstly, we experiment through the first balloon injury of rabbit iliac artery to establish diabetic AS model, and then based on the PTA operation, better simulate the process of restenosis in diabetic patients with lower extremity arteriosclerosis after PTA surgery, double balloon injury / surgery application for the expansion of restenosis on the establishment of animal model is more accurate. Next, we observed the progress and characteristics of restenosis after PTA after observing the stenosis rate, intimal hyperplasia and distribution of VSMCs in different time points after PTA. Third, we observed the effects of statins on the migration, proliferation and restenosis degree of vascular smooth muscle cells, so as to determine whether they could inhibit the occurrence of restenosis after PTA. Methods 1. Model and group selected male New Zealand white rabbits (weight about 1.7kg). After 1 weeks of adaptive feeding, high fat diet was given to the experiment to terminate. 1, double balloon injury / dilation and restenosis model was established. (1) diabetes model was established: four yuan (four 80mg/kg) was injected into the ear vein, and blood glucose was measured after 1 weeks to confirm the establishment of the diabetes model. (2) AS model: 1 weeks after Duiqia artery balloon injury was established by diabetes model, strain 4 weeks after surgery by ultrasound examination confirmed the iliac artery AS model. (3) restenosis model was established: a restenosis model was established by PTA operation at the vascular site of AS. 2. Animal models of restenosis were randomly divided into four groups: 7 day restenosis group, 14 day restenosis group, 28 day restenosis group, 28 days atorvastatin group, and another control group. The treatment of each group (1) 7 days restenosis group: the same volume of saline was given to the stomach and was executed seventh days after PTA. (2) 14 day restenosis group: the same volume of normal saline was given to the stomach, and fourteenth days after PTA was executed (3) for 28 days. The restenosis group was given the same volume of normal saline, and PTA was executed twenty-eighth days later. (4) the 28 day atorvastatin group: the routine dose of atorvastatin (2.5mg/kg/d) was given to the stomach for 28 days. (5) the control group: the same surgical operation was performed in the animals of the restenosis model group, but the balloon dilation was not carried out, and the same volume of physiological saline was given to the stomach. Two. Specimens were collected and tested for indicators 1. According to the experimental grouping, animals were killed at different time points (seventh days, 14 days, 28 days after PTA), and blood samples and iliac artery specimens were left. 2, detection of serum lipids: (1) serum total cholesterol (TC); (2) triglyceride (Triglyceride, TG); (3) low density lipoprotein (low density lipoprotein cholesterin; LDL-C); (4) high density lipoprotein (high, TG). 3, detection of vascular morphology: iliac artery segment was fixed in 4% paraformaldehyde, dehydrated after embedding in paraffin sections, the following test (1) the general morphological observation of blood vessels stained by HE (2) to observe the morphology and collagen vascular smooth muscle stained by Masson (3) to observe the elastic fiber morphology by EVG staining. (4) using Image-Pro Plus 6 image analysis software to measure vascular plaque area, intima and middle membrane area to calculate vessel stenosis rate and intima / middle membrane ratio. 4. Immunohistochemical analysis: the number and differentiation of intimal and mesangial proliferating cells were measured by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA).
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R587.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 Jing ZHAO;Hui-min YAN;Ya LI;Jia WANG;Lu HAN;Zhi-hao WANG;Meng-xiong TANG;Wei ZHANG;Yun ZHANG;Ming ZHONG;;匹伐他汀鈣對高膽固醇血癥患者動脈粥樣硬化的影響（英文）[J];Journal of Zhejiang University-Science B(Biomedicine & Biotechnology);2015年05期

2 童保文;林志鴻;謝良地;許昌聲;;兔頸動脈球囊損傷后血小板聚集和活化及阿托伐他汀的作用[J];南方醫(yī)科大學(xué)學(xué)報;2014年08期

3 Ya-fei SHI;Ju-fang CHI;Wei-liang TANG;Fu-kang XU;Long-bin LIU;Zheng JI;Hai-tao LV;Hang-yuan GUO;;Effects of rosuvastatin on the production and activation of matrix metalloproteinase-2 and migration of cultured rat vascular smooth muscle cells induced by homocysteine[J];Journal of Zhejiang University-Science B(Biomedicine & Biotechnology);2013年08期

4 潘軼斌;包麗芳;陳杭;金烈;張向陽;華崇俊;;不同劑量瑞舒伐他汀對急性冠脈綜合征患者血NF-κB,sICAM-1表達(dá)的影響[J];中國現(xiàn)代應(yīng)用藥學(xué);2011年06期

5 徐戈;周勝華;蔣路平;孫智山;;阿托伐他汀對IL-1β誘導(dǎo)的大鼠VSMC中Cat S和NF-κB表達(dá)的影響[J];廣西醫(yī)科大學(xué)學(xué)報;2010年01期

6 任天舒;李林鵬;姜遠(yuǎn)英;;高膽固醇血癥大鼠自由基代謝紊亂及阿托伐他汀的干預(yù)作用[J];藥學(xué)實踐雜志;2007年03期

，

本文編號：1341909