本文選題：動脈粥樣硬化　切入點：清脂通脈顆粒　出處：《遼寧中醫(yī)藥大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過觀察清脂通脈顆粒對動脈粥樣硬化(AS)模型家兔的血脂代謝及膽固醇逆轉(zhuǎn)運受體SR-BI mRNA和蛋白表達(dá)方面的影響,深入研究在防治動脈粥樣硬化方面清脂通脈顆粒的作用機制。材料與方法:1.本實驗選取的實驗動物為健康雄性家兔,在實驗中的動物的分組均采取隨機數(shù)字表的方法,我們將22只家兔,隨機選取6只為空白組,剩余16只為造模組。使各組家兔每天都自由飲水,室溫保持在20?C-25?C,適應(yīng)性飼養(yǎng)一周后,對模型組進(jìn)行造模,造模方法即采用傳統(tǒng)的高脂飼料喂養(yǎng)加維生素D3注射法進(jìn)行動脈粥樣硬化模型的復(fù)制。模型組將維生素D3一次性注射于腹腔,注射劑量為(60萬IU/kg)�？瞻捉M給予正常飼料顆粒喂養(yǎng),并同樣適應(yīng)性飼養(yǎng)一周后,在實驗第一天在腹腔注射同等劑量的生理鹽水。高脂飼料喂養(yǎng),時間為4周,從空白組和模型組各隨機抽取1只家兔進(jìn)行頸動脈病理切片,采用HE染色法觀察頸動脈粥樣斑塊病理改變,評價造模成功與否。造模成功后,按照隨機數(shù)字表法將造模組15只家兔,按照每組5只隨機分為模型組、辛伐他汀組(西藥組)、清脂通脈顆粒組(中藥組)然后開始對辛伐他汀組(西藥組)、清脂通脈顆粒組(中藥組)進(jìn)行藥物治療,兩組分別每日一次經(jīng)口灌飼辛伐他汀、清脂通脈顆粒,同樣連續(xù)灌飼4周。2.至實驗第9周末禁食,對各組家兔進(jìn)行經(jīng)耳緣靜脈采集血液樣本,對血清進(jìn)行分離,目的是檢測各組血清中血脂四項指標(biāo),分別包括總膽固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)和高密度脂蛋白(HDL-C)的含量采用全自動生化儀,并檢測各組家兔氧化型低密度脂蛋白(OX-LDL)的含量通過酶聯(lián)免疫吸附法(ELISA)。至實驗9周末禁食經(jīng)耳緣靜脈麻醉后處死動物,剖開腹部,摘取肝臟尾狀葉,檢測各組動物肝臟組織中SR-BImRNA及SR-BI蛋白的表達(dá)水平分別通過RT-PCR方法和Western-blot方法。3.對四組實驗數(shù)據(jù)進(jìn)行統(tǒng)計分析,分析軟件采用SPSS17.0,其中各組的計量資料以均數(shù)±標(biāo)準(zhǔn)差((?)±S)表示,多個組間的數(shù)據(jù)進(jìn)行比較運用單因素方差分析,統(tǒng)計結(jié)果以P0.01有統(tǒng)計學(xué)意義,P0.05則無統(tǒng)計學(xué)意義。結(jié)果:1.血脂四項檢測結(jié)果得出:與空白組比較,模型組、西藥組、中藥組TG、TC、LDL-C含量均升高(P0.01);與模型組比較,西藥組,中藥組TG、TC、LDL-C含量均降低(P0.01);與西藥組比較,中藥組TG、TC、LDL-C含量均降低(P0.05);與空白組比較,模型組、西藥組、中藥組HDL-C含量均降低(P0.01);與模型組比較,西藥組、中藥組HDL-C含量均升高(P0.01);與西藥組比較,中藥組HDL-C含量降低(P0.05)。2.氧化型低密度脂蛋白檢測結(jié)果:與空白組比較,模型組、西藥組、中藥組OX-LDL含量均升高(P0.01);與模型組比較,西藥組、中藥組OX-LDL含量均降低(P0.01);與西藥組比較,中藥組OX-LDL含量升高(P0.05)。3.Western-blot結(jié)果顯示:SR-BI蛋白的目的條帶大約在36k Da處,統(tǒng)計積分光密度值的結(jié)果可以得出,與空白組比較,模型組、西藥組、中藥組SR-BI表達(dá)降低(p0.01)。與模型組比較,西藥組、中藥組SR-BI表達(dá)升高(p0.01)。與西藥組比較,無顯著差異(p0.05)。4.RT-PCR結(jié)果顯示:SR-BI mRNA的目的條帶大約在457bp處,統(tǒng)計積分光密度值的結(jié)果可以得出,與空白組比較,模型組、西藥組、中藥組SR-BImRNA表達(dá)降低(p0.01)。與模型組比較,西藥組、中藥組SR-BImRNA表達(dá)升高(p0.01)。與西藥組比較,無顯著差異(p0.05)。結(jié)論:1.通過高脂飼料喂養(yǎng)家兔,發(fā)現(xiàn)模型組中含量顯著升高的有TC、TG、LDL-C及OX-LDL,含量顯著降低的為HDL-C,說明高脂血癥動脈硬化兔模型造模成功。2.清脂通脈顆粒使動脈粥樣硬化模型家兔肝細(xì)胞組織中SR-BI mRNA的表達(dá)升高。3.清脂通脈顆粒使動脈粥樣硬化模型家兔肝細(xì)胞組織中SR-BI蛋白的表達(dá)升高。4.清脂通脈顆粒實現(xiàn)其對膽固醇的逆向轉(zhuǎn)運是通過上調(diào)肝細(xì)胞中SR-BI mRNA及蛋白的表達(dá)。5.肝細(xì)胞中SR-BI mRNA及蛋白的表達(dá)升高,清脂通脈顆粒防治動脈粥樣硬化的可能機制是通過促進(jìn)膽固醇逆轉(zhuǎn)運實現(xiàn)。
[Abstract]:Objective: through the observation of Qingzhi Tongmai Granule on atherosclerosis (AS) metabolism and cholesterol transport reversal rabbit model of receptor SR-BI mRNA and protein expression of the influence of in-depth study of the mechanism of Qing fat Tongmai granule in treating atherosclerosis. Materials and methods: the experimental animal experiment for the selected 1. healthy male rabbits. The packet in the experiment animal adopt the method of random number table, we will be 22 rabbits, 6 rabbits were randomly selected as control group, the remaining 16 rats were model group. The rabbits of each group every day free drinking water, room temperature maintained at 20? C-25? C, afteradaptivebreedingforoneweek, the model group was made die, mold method that using the traditional model of atherosclerosis in high fat diet with vitamin D3 injection in the model group. The replication of vitamin D3 injection in abdominal cavity injection (600 thousand IU/kg) blank. Group with normal feed particle feeding, and the same afteradaptivebreedingforoneweek, saline in the first day in the same dose intraperitoneal injection. High fat diet for 4 weeks, from the blank group and the model group were randomly selected 1 Rabbits Carotid artery pathological sections, the pathological changes of carotid atherosclerotic plaque HE to observe the staining, the evaluation model is successful or not. After the success of the model, according to the random number table method model of 15 rabbits, with 5 rats in each group were randomly divided into model group, simvastatin group (Western medicine group), Qingzhi Tongmai granule group (Chinese medicine group) and simvastatin group (Western medicine group) on Qingzhi, Tongmai granule group (Chinese medicine group) for drug treatment, two groups were once daily oral intragastric administration of simvastatin, Qingzhi Tongmai Granules, the same continuous feeding of 4 weeks.2. to the ninth weekend fasting by ear vein blood samples were collected and analyzed in each group, Separation of serum, the aim is to detect four indexes of blood lipid in serum, including total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) content by automatic biochemical analyzer, and were observed by oxidation of low density lipoprotein (the content of OX-LDL) by enzyme linked immunosorbent assay (ELISA). To the 9 weekend of fasting by ear vein anesthesia after the death of animal, open abdomen, liver caudate lobe, the expression level of SR-BImRNA and SR-BI protein in liver tissue were detected in animal respectively by RT-PCR method and Western-blot method of.3. of four groups of experimental data statistics the analysis, using SPSS17.0 analysis software, the measurement data were the mean standard deviation ((?) + S) said that many groups of data were compared using single factor analysis of variance, statistical results in P0.01 have statistical significance, P0. 05鍒欐棤緇熻瀛︽剰涔，

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