發(fā)布時(shí)間：2018-05-05 05:22

  本文選題：參草通脈顆粒 + klotho�。� 參考：《遼寧中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：目的:通過(guò)建立慢性心衰(CHF)大鼠模型,觀察參草通脈顆粒對(duì)慢性心衰大鼠心肌及腎組織klotho的影響以及心肌組織結(jié)構(gòu)的改變。圍繞參草通脈顆粒對(duì)大鼠模型慢性心衰癥狀的作用機(jī)理和作用靶點(diǎn),分別從基因、蛋白和組織形態(tài)學(xué)等渠道發(fā)現(xiàn)和探索將中醫(yī)藥防治作為治療慢性心衰重要手段的科學(xué)依據(jù)。材料與方法:1.通過(guò)對(duì)實(shí)驗(yàn)大鼠采取結(jié)扎冠狀動(dòng)脈前降支造成局部心梗,再對(duì)大鼠進(jìn)行游泳及減食等方法造成慢性心衰模型,后觀察大鼠狀態(tài),且經(jīng)多普勒超聲心動(dòng)圖檢測(cè)射血分?jǐn)?shù)(EF)≤55%或CI≤180ml/min/kg,判定慢性心衰大鼠模型造模是否成功。2.將建立的慢性心衰大鼠模型分4組培養(yǎng)。首先,設(shè)立假手術(shù)組模型用于后期對(duì)比研究,該組模型不采取任何藥物干預(yù),僅用常規(guī)等容積蒸餾水灌胃。其次,保留一組心衰大鼠模型為模型組,該組采取同假手術(shù)組培養(yǎng)方式。第三,設(shè)立西藥組采取灌服賴(lài)諾普利干預(yù)。最后,設(shè)立中藥組模型,灌服參草通脈顆粒,中藥組按照9.2g生藥/Kg/d灌服參草通脈顆粒,賴(lài)諾普利西藥組1.5mg生藥/Kg/d。預(yù)計(jì)培養(yǎng)4周時(shí)間,隨后經(jīng)大鼠尾靜脈采血后,麻醉處死全部大鼠,取其大鼠心臟心肌組織及腎組織。3.將所取各組樣本分別通過(guò)ELISA法、蛋白免疫印跡技術(shù)和實(shí)時(shí)熒光定量PCR技術(shù)收集相關(guān)數(shù)據(jù),主要對(duì)比不同組模型間BNP濃度,心肌及腎組織klotho的變化情況及電鏡觀察各組間心肌細(xì)胞組織形態(tài)學(xué)變化情況。結(jié)果:1.慢性心衰大鼠BNP濃度與假手術(shù)組比較,治療組(中藥組,西藥組)及模型組均升高,p0.05。其中假手術(shù)組BNP濃度為450±37.03pg/ml,模型組BNP濃度1017.55±98.6pg/ml,中藥組與西藥組BNP濃度分別為681.08±75.31pg/ml,686.53±74.86pg/ml,p0.05。2.慢性心衰大鼠模型組心肌klotho mRNA表達(dá)明顯降低,為0.0003±0.00004(與假手術(shù)組比較P0.01);西藥組樣本心肌klotho mRNA具體數(shù)值為0.0020±0.0009相較于中藥組(具體數(shù)值為0.0012±0.0006)p0.05,組間比較治療組與模型組P0.01與假手術(shù)組P0.01。3.慢性心衰大鼠模型組心肌klotho蛋白表達(dá)明顯下降,為0.0291±0.0025(與假手術(shù)組比較P0.01);西藥組樣本心肌klotho蛋白具體數(shù)值為0.2475±0.1407,相較于中藥組(具體數(shù)值為0.215±0.077)p0.05,組間比較治療組與模型組P0.01與假手術(shù)組P0.01。4.慢性心衰模型組腎組織klotho mRNA表達(dá)明顯降低,為0.0022±0.0007,(與假手術(shù)組比較P0.05);西藥組樣本腎klotho mRNA具體數(shù)值為0.0067±0.00209,相較于中藥組(具體數(shù)值為0.0063±0.0022)p0.05,組間比較治療組與模型組P0.01與假手術(shù)組P0.01。5.慢性心衰模型組腎組織klotho蛋白表達(dá)明顯降低,為0.0639±0.0203(與假手術(shù)組比較P0.01);西藥組樣本腎klotho蛋白具體數(shù)值為0.2150±0.0303,相較于中藥組(數(shù)值為0.2057±0.0457)p0.05,組間比較治療組與模型組P0.01與假手術(shù)組P0.01。6.假手術(shù)組,可見(jiàn)完整的心肌細(xì)胞,肌纖維整齊有序,呈均勻清晰的Z線(xiàn)排列,肌節(jié)可見(jiàn)明顯的周期性,心肌組織間質(zhì)無(wú)水腫,大量橢圓形線(xiàn)粒體排列整齊于心肌肌纖維中。模型組,肌節(jié)結(jié)構(gòu)不清或消失,即肌原纖維斷裂、溶解,肌節(jié)紊亂,心肌組織間質(zhì)見(jiàn)明顯水腫,壞死區(qū)域面積大,壞死嚴(yán)重,線(xiàn)粒體形態(tài)不一,見(jiàn)腫脹、破裂或消失,空泡化嚴(yán)重。中藥組,肌節(jié)較清晰,仍有局部肌纖維斷裂、溶解,線(xiàn)粒體排列于肌絲之間,呈輕度水腫,尚有部分線(xiàn)粒體空化。西藥組,肌節(jié)較清晰,部分肌纖維斷裂、溶解,線(xiàn)粒體排列于肌絲之間,輕度腫脹,且空化線(xiàn)粒體較少。結(jié)論:1.通過(guò)對(duì)大鼠模型超聲心動(dòng)圖檢測(cè),并且輔以對(duì)比健康大鼠行為,證明利用結(jié)扎冠狀動(dòng)脈前降支配合減食與力竭式游泳可成功造成大鼠心衰模型。2.通過(guò)4周參草通脈顆粒的治療后利用ELISA法檢測(cè)BNP濃度下降,通過(guò)實(shí)時(shí)熒光定量PCR、Western blot檢測(cè)大鼠心肌組織,發(fā)現(xiàn)參草通脈顆粒對(duì)心肌組織klotho mRNA與蛋白有明顯升高其表達(dá)的作用;并且通過(guò)電鏡觀察心肌組織證明參草通脈顆�？善鸬揭种苹蚰孓D(zhuǎn)心室重構(gòu)的作用。3.通過(guò)4周參草通脈顆粒的治療后利用實(shí)時(shí)熒光定量PCR、Western blot檢測(cè)大鼠腎組織,發(fā)現(xiàn)參草通脈顆粒對(duì)腎組織klothomRNA與蛋白有明顯升高其表達(dá)的作用;參草通脈顆粒可起到抑制腎損傷的作用。
[Abstract]:Objective: by establishing a rat model of chronic heart failure (CHF), we observed the effect of Shen Cao Tongmai Granule on the Klotho of myocardium and kidney tissue in chronic heart failure rats and the changes of the structure of myocardium. The mechanism and target points of Shen Cao Tong Mai Granule on the symptoms of chronic heart failure in rat model were discussed from gene, protein and histomorphology. We found and explored the scientific basis of traditional Chinese medicine as an important means to treat chronic heart failure. Materials and methods: 1. by ligating the anterior descending branch of the coronary artery to cause local myocardial infarction in experimental rats, and then swimming and reducing food in rats, the model of chronic heart failure was created, and then the state of rats was observed, and the Doppler echocardiography was used to examine the rats. The score of ejection fraction (EF) was less than 55% or CI < 180ml/min/kg. To determine whether the model of chronic heart failure rat model was successful or not, the model of chronic heart failure in.2. would be divided into 4 groups. First, the model of sham operation group was set up for the later contrast study. The model group of heart failure rats was the model group, and the group adopted the same group as the sham operation group. Third, the western medicine group was set up to take the intervention of lisinopril. Finally, the Chinese medicine group was set up and filled with Shen Cao Tongmai granule. The Chinese medicine group was filled with Shen Cao Tong Mai Granule according to the 9.2g raw medicine /Kg/d, and the /Kg/d. of the 1.5mg medicine of lisinopixi drug group was expected to be cultivated for 4 weeks. After the rat tail vein was collected, all rats were killed, and the cardiac muscle tissue and.3. of the rats were taken to collect all the samples by ELISA, protein immunoblotting and real-time fluorescence quantitative PCR. The concentration of BNP and the changes of Klotho in the myocardium and kidney tissue were compared. Results: 1. the concentration of BNP in the chronic heart failure rats was higher than that in the sham group. The treatment group (Chinese medicine group, western medicine group) and the model group were all increased, and the concentration of BNP in the sham operation group was 450 + 37.03pg/ml, the concentration of BNP in the model group was 1017.55 + 98.6pg/ml, and the concentration of BNP in the Chinese medicine group and the western medicine group was respectively the concentration of BNP in the Chinese medicine group and the western medicine group. 681.08 + 75.31pg/ml, 686.53 + 74.86pg/ml, the expression of Klotho mRNA in the rat model group of p0.05.2. chronic heart failure was significantly reduced, 0.0003 + 0.00004 (compared with the sham operation group P0.01), and the specific value of Klotho mRNA in the western medicine group was 0.0020 + 0.0009 compared to the Chinese medicine group (with the body value of 0.0012 + 0.0006) P0.05, the comparison between the group and the model group was compared with the model group. The expression of Klotho protein in the myocardium of the model group P0.01 and the sham group P0.01.3. was 0.0291 + 0.0025 (compared with the sham operation group P0.01), and the specific value of Klotho protein in the western medicine group was 0.2475 + 0.1407, compared with the traditional Chinese medicine group (the specific value was 0.215 + 0.077) P0.05, and the comparative treatment group and the model group P0.01 were compared. The expression of Klotho mRNA in renal tissue of P0.01.4. chronic heart failure model group was significantly lower than that in sham group, which was 0.0022 + 0.0007 (compared with sham operation group P0.05), and the specific value of Klotho mRNA in the sample kidney of Western medicine group was 0.0067 + 0.00209, compared with the traditional Chinese Medicine group (the specific value was 0.0063 + 0.0022) P0.05, and the comparison group was compared with the model group P0.01 and the sham operation group P. In 0.01.5. chronic heart failure model group, the expression of Klotho protein in renal tissue decreased significantly, which was 0.0639 + 0.0203 (compared with sham operation group P0.01). The specific value of Klotho protein in the western medicine group was 0.2150 + 0.0303, compared with the traditional Chinese medicine group (0.2057 + 0.0457) P0.05, and the comparison treatment group and the model group P0.01 and the sham operation group P0.01.6. sham operation group, The complete myocardial cells were seen, the muscle fibers were orderly and orderly, the Z lines were arranged in a uniform and clear line, the myofibrils were obvious periodicity, the interstitial tissue of the myocardium was not edema, and a large number of oval mitochondria were arranged neatly in the muscle fibers. The model group, the myofibril structure was not clear or disappeared, that is, the myofibril rupture, dissolving, the myofibril disorder, and the interstitial tissue of the myocardium. Edema, necrotic area large area, necrosis serious, mitochondria form different, see swelling, rupture or disappearance, vacuolization serious. The Chinese medicine group, the myojoints are clear, there are still local muscle fiber fracture, dissolve, the mitochondria are arranged between the myes, with a mild edema, the western medicine group, the myojoints are clearer, some muscle fibers break and dissolve. The mitochondria were arranged between the muscle fibers, slightly swelling and the cavitation mitochondria were less. Conclusion: 1. through the echocardiographic examination of the rat model and the behavior of the healthy rats, it is proved that the model.2. of the rat heart failure model can be successfully made through the ligature of the anterior descending branch of the coronary artery and the exhaustive swimming of the rats through 4 weeks. After treatment, ELISA method was used to detect the decrease of BNP concentration. The myocardial tissue of rats was detected by real-time fluorescence quantitative PCR and Western blot. The effect of Shen Cao Tong Mai Granule on the expression of Klotho mRNA and protein was found in the myocardium, and the myocardial tissue was observed by electron microscope to prove that the ginseng Tongmai granule could inhibit or reverse the ventricular remodeling. The effect of.3. on renal tissue of rats was detected by real-time fluorescence quantitative PCR and Western blot after 4 weeks of ginseng Tongmai Granules. The effect of Shen Cao Tong Mai Granule on the expression of klothomRNA and protein in renal tissue was found, and the effect of Shen Cao Tongmai Granules could inhibit the renal injury.

【學(xué)位授予單位】：遼寧中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R285.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 方鵬飛;常麗霞;宋淵;;三七總皂苷臨床應(yīng)用研究進(jìn)展[J];中醫(yī)藥學(xué)報(bào);2016年03期

2 馬金;張艷;;慢性心力衰竭中醫(yī)病機(jī)“氣虛-血瘀-水停”與“心室重構(gòu)”的相關(guān)性探討[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2016年17期

3 謝澤娟;鐘曉珊;陳嘉迪;;運(yùn)動(dòng)康復(fù)護(hù)理對(duì)老年慢性心衰患者的影響[J];中國(guó)實(shí)用醫(yī)藥;2016年03期

4 張新鋒;楊慶坤;;他汀類(lèi)藥物治療慢性心力衰竭的利弊探討[J];醫(yī)學(xué)理論與實(shí)踐;2015年22期

5 洪華山;;老年慢性心力衰竭的藥物和非藥物治療現(xiàn)狀[J];中華老年心腦血管病雜志;2015年08期

6 胡茜;黃小春;;淺議益腎活血法治療心力衰竭之理論依據(jù)[J];江蘇中醫(yī)藥;2015年08期

7 巫朝倫;;淺談洋地黃類(lèi)藥物中毒[J];世界最新醫(yī)學(xué)信息文摘;2015年55期

8 張軍;代文靜;周敬群;張家俊;曹志剛;;抗衰老Klotho蛋白減輕大鼠乳鼠心肌細(xì)胞缺氧/復(fù)氧損傷[J];中國(guó)病理生理雜志;2015年06期

9 寶源然;羅紅英;;慢性心功能不全患者利尿劑抵抗的原因分析[J];中西醫(yī)結(jié)合心血管病電子雜志;2015年15期

10 馬燕;巴·巴音斯勒瑪;郭李平;王娟;呂忠英;韓素霞;;BNP監(jiān)測(cè)指導(dǎo)慢性心衰患者早期應(yīng)用β-受體阻滯劑的效果觀察[J];中西醫(yī)結(jié)合心血管病電子雜志;2015年11期

相關(guān)博士學(xué)位論文 前2條

1 金鑫;血管緊張素Ⅱ通過(guò)AT1R/Rho/ROCK/JNK通路上調(diào)肥大心肌細(xì)胞分泌型卷曲相關(guān)蛋白5的表達(dá)[D];河北醫(yī)科大學(xué);2015年

2 唐榮;Klotho基因在高血壓腎損害腎小管上皮細(xì)胞凋亡中的作用及冬蟲(chóng)夏草對(duì)其的影響[D];中南大學(xué);2009年

相關(guān)碩士學(xué)位論文 前3條

1 賈政;Klotho基因與大鼠心力衰竭心肌重構(gòu)的相關(guān)性研究[D];桂林醫(yī)學(xué)院;2015年

2 張洋;益氣活血復(fù)方對(duì)慢性心衰大鼠心�、瘛ⅱ笮湍z原表達(dá)影響的實(shí)驗(yàn)研究[D];遼寧中醫(yī)藥大學(xué);2011年

3 禮海;益氣活血復(fù)方對(duì)慢性心衰大鼠心肌MMP-1、TIMP-1影響相關(guān)性及抗心室重構(gòu)機(jī)制的研究[D];遼寧中醫(yī)藥大學(xué);2010年

，

本文編號(hào)：1846394