本文關(guān)鍵詞： IgA腎病 RAS系統(tǒng) 血管緊張素Ⅱ（AngⅡ） 血管緊張素ⅡⅠ型受體（AT1） Ⅳ型膠原 腎小球硬化　出處：《河北醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：IgA腎病（IgAN）是世界范圍內(nèi)常見的原發(fā)性腎小球疾病，近年來研究發(fā)現(xiàn)15%～20%的患者在確診后10年內(nèi)、30%～35%的患者在確診后20年內(nèi)進(jìn)入終末期腎衰竭,嚴(yán)重威脅著患者的身心健康。IgAN在我國(guó)發(fā)病率較高，其臨床表現(xiàn)多樣，以反復(fù)發(fā)作性血尿?yàn)橹饕攸c(diǎn)，可伴有蛋白尿、高血壓等。IgAN的診斷目前主要是依靠病理檢查并結(jié)合血尿、蛋白尿等臨床表現(xiàn)，其病理特征主要包括腎小球系膜區(qū)多聚IgA1分子伴少量C3、IgM、IgG沉積及系膜細(xì)胞增殖、系膜基質(zhì)的增多。關(guān)于其發(fā)病機(jī)制仍然知之甚少，許多學(xué)者認(rèn)為其是多種因素綜合作用的結(jié)果。近年來，細(xì)胞因子的作用引起人們的關(guān)注，研究發(fā)現(xiàn)腎臟局部的RAS系統(tǒng)激活，可以刺激系膜細(xì)胞增殖、系膜基質(zhì)增寬及細(xì)胞外基質(zhì)的合成，在IgAN的進(jìn)展過程中發(fā)揮重要作用。目前IgAN的治療僅限于對(duì)癥支持及免疫抑制等治療，因此發(fā)現(xiàn)有效的特異性治療方案成為研究熱點(diǎn)。因此我們提出“益氣養(yǎng)陰法”治療IgAN，應(yīng)用于臨床上后取得了不錯(cuò)的治療效果，但其具體的治療作用機(jī)制仍然不是很清楚。 本文首先建立IgAN動(dòng)物模型，應(yīng)用益氣養(yǎng)陰中藥按照相應(yīng)的藥物劑量進(jìn)行干預(yù)治療。通過觀察大鼠24小時(shí)尿蛋白定量、尿紅細(xì)胞計(jì)數(shù)、腎功能等，同時(shí)檢測(cè)血清中AngⅡ的水平及腎組織AT1、Ⅳ型膠原的表達(dá)情況，旨在探討益氣養(yǎng)陰中藥治療IgA腎病的作用機(jī)制及藥物作用靶點(diǎn)。 方法：40只雄性清潔級(jí)SD大鼠被選入實(shí)驗(yàn)。我們將這40只大鼠隨機(jī)分成正常對(duì)照組10只、模型組10只、貝那普利組10只、中藥組10只。除正常對(duì)照組外的后三組均運(yùn)用BSA、脂多糖、蓖麻油、四氯化碳復(fù)制IgAN大鼠動(dòng)物模型。具體方法如下：將免疫原BSA用蒸餾水配制成100g/L濃度，隔天灌胃，劑量為400mg/kg，持續(xù)8周；每周一次皮下注射四氯化碳0.1ml、蓖麻油0.3ml，連續(xù)9周；并于第六周尾靜脈注射脂多糖0.05mg。正常對(duì)照組給予相同劑量的蒸餾水隔日灌胃8周；每周一次皮下注射生理鹽水0.4ml；第六周尾靜脈注射生理鹽水0.2ml。然后從第10周起，貝那普利組、中藥組分別予以鹽酸貝那普利、中藥灌胃，，正常對(duì)照組、模型組予以等量生理鹽水灌胃，持續(xù)8周。實(shí)驗(yàn)過程中，我們將大鼠置于代謝籠內(nèi)分別于實(shí)驗(yàn)的各個(gè)時(shí)間點(diǎn)留取尿液標(biāo)本，測(cè)定尿紅細(xì)胞計(jì)數(shù)及24小時(shí)尿蛋白定量。實(shí)驗(yàn)第17周末，麻醉動(dòng)物，留取血液標(biāo)本，迅速采集大鼠腎臟標(biāo)本。血液標(biāo)本離心后用半自動(dòng)生化儀測(cè)定大鼠腎功能及放射免疫法測(cè)定血清中AngⅡ的水平。最后，于光鏡、電鏡下觀察大鼠腎臟組織病理形態(tài)學(xué)改變，免疫熒光法檢測(cè)大鼠腎臟IgA免疫復(fù)合物沉積，免疫組織化學(xué)技術(shù)檢測(cè)Ⅳ型膠原的表達(dá)情況，RT-PCR技術(shù)檢測(cè)AT1的基因表達(dá)水平。 結(jié)果: 模型組、貝那普利組、中藥組大鼠ATI基因表達(dá)較正常組明顯升高（P0.01），貝那普利組、中藥組大鼠這個(gè)指標(biāo)的表達(dá)較模型組有所降低（P0.01），貝那普利組、中藥組間無明顯區(qū)別。 正常組大鼠Ⅳ型膠原主要在腎小球及腎小管基底膜表達(dá)，系膜區(qū)、腎間質(zhì)幾乎不表達(dá)；模型組、貝那普利組、中藥組大鼠較正常組Ⅳ型膠原的表達(dá)明顯增多，可見其在系膜區(qū)表達(dá)明顯升高；貝那普利組、中藥組較模型組表達(dá)較模型組降低，兩者之間無明顯差異。 結(jié)論: 1益氣養(yǎng)陰中藥可以降低IgA腎病大鼠尿蛋白及尿紅細(xì)胞計(jì)數(shù)，從而保護(hù)腎功能； 2益氣養(yǎng)陰中藥可以顯著改善IgA腎病大鼠腎小球上皮細(xì)胞足突的融合、系膜細(xì)胞的增生及系膜基質(zhì)的增寬等病理損害； 3模型組大鼠血清中AngⅡ水平、腎臟局部AT1mRNA的表達(dá)明顯增加，提示腎臟局部RAS系統(tǒng)的激活在IgAN的進(jìn)展中起重要作用。益氣養(yǎng)陰中藥可能是通過阻斷這一途徑而達(dá)到對(duì)IgA腎病大鼠治療作用的； 4IgA腎病大鼠腎組織AT1mRNA的表達(dá)與Ⅳ型膠原的合成呈正相關(guān)，提示RAS系統(tǒng)的激活可以促進(jìn)細(xì)胞外基質(zhì)的合成。益氣養(yǎng)陰中藥可以顯著抑制這一作用，延緩疾病的進(jìn)展。
[Abstract]:Objective: IgA nephropathy (IgAN) is a worldwide common primary glomerular disease, recent studies have found that 15% to 20% of patients in the 10 years after diagnosis, 30% ~ 35% of the patients in the 20 years after diagnosis in end-stage renal failure, a serious threat to physical and mental health of patients with.IgAN incidence in our country high rate of clinical manifestations with recurrent hematuria as the main character, may be associated with proteinuria, hypertension and other.IgAN diagnosis mainly depends on pathological examination combined with hematuria, proteinuria, the major pathological features including mesangial poly IgA1 molecules with a small amount of C3, IgM, IgG, proliferation deposition and mesangial cells, increased mesangial matrix. The pathogenesis is still poorly understood, many scholars believe that it is the result of many factors. In recent years, the role of cytokines to cause the attention of people, the study found that kidney Bureau The activation of RAS system, can stimulate the proliferation of mesangial cells and mesangial matrix synthesis and broadening of the extracellular matrix, play an important role in the progression of IgAN. The treatment of IgAN is limited to symptomatic and immunosuppressive therapy, so that specific therapy effective scheme so we become a research hotspot. Put forward the "supplementing Qi and nourishing Yin" in the treatment of IgAN, the application has achieved good curative effect in clinic, but the treatment mechanism is still not very clear.
In this paper, we establish the IgAN animal model, application of Tonifying Qi and nourishing Yin were treated according to the drug dosage accordingly. Through the observation of the rat 24 hour urinary protein, urine red blood cell count, renal function, and serum levels of Ang II and renal tissue to detect AT1 expression of type IV collagen, mechanism and medicine the target to explore Yiqiyangyin medicine for the treatment of IgA nephropathy.
Methods: 40 male SD rats were selected in the experiment. The 40 rats were randomly divided into normal control group 10 rats, 10 rats in model group, Benner Pury group 10, Chinese medicine group 10. Except the normal control group after three groups using BSA, lipopolysaccharide, castor oil, preparation IgAN rat animal model of complex carbon tetrachloride. The method is as follows: the immunogenicity of BSA diluted in distilled water 100g/L concentration, the intragastric dose of 400mg/kg for 8 weeks; weekly subcutaneous injection of carbon tetrachloride 0.1ml, castor oil, 0.3ml, for 9 weeks; and the control group was given the same dose of distilled water by gavage every day 8 weeks of normal at sixth weeks of intravenous injection of lipopolysaccharide 0.05mg.; weekly subcutaneous injection of saline 0.4ml; sixth weeks of intravenous injection of saline 0.2ml. and then from the tenth week, Benner Pury group, Chinese medicine group were given intragastric administration of Benner Pury hydrochloride, Chinese medicine, normal control Group and model group were given normal saline, lasted 8 weeks. During the experiment, we the rats were placed in metabolism cages at the each time point of urine samples, determination of urine red blood cell count and urine protein in 24 hours. At the end of the seventeenth week experiment, anesthesia animal blood samples rapidly collecting samples of rat kidney. The level of blood serum Ang determination of renal function and radioimmunoassay for rat samples after centrifugation by semi-automatic biochemical analyzer. Finally, in addition, the pathological changes of kidney tissue were observed under electron microscope, detection of renal IgA in rats immune complex deposition immunofluorescence, expression of IV type collagen immunohistochemistry, detect the expression of AT1 RT-PCR gene.
Result:
The expression of ATI gene in model group, Benner Pury group and Chinese medicine group was significantly higher than that in the normal group (P0.01). The expression of this index in the Benner Pury group and the traditional Chinese medicine group was lower than that in the model group (P0.01), and there was no significant difference between the Benner Pury group and the Chinese medicine group.
The rats in the normal group of type IV collagen is mainly expressed in renal tubular and glomerular basement membrane and mesangial region, renal interstitial almost no expression; model group, Benner Pury group, the expression of type IV collagen than normal group rats increased significantly, the expression was significantly increased in the mesangium; benazepril group. The Chinese medicine group than in the model group expressed lower than model group, no significant difference between the two.
Conclusion:
1 supplementing qi and nourishing Yin can reduce the urine protein and urine red blood cell count of IgA nephropathy rats, and protect the renal function.
2, Chinese herbs for supplementing qi and nourishing Yin can significantly improve the fusion of glomerular epithelial cells, foot processes, proliferation of mesangial cells and widening of mesangial matrix in IgA nephropathy rats.
3, the level of Ang II in serum and the expression of AT1mRNA in kidney of rats in model group increased significantly, suggesting that activation of RAS system in kidney plays an important role in the progress of IgAN. Yiqi Yangyin herbs may play a role in the treatment of IgA nephropathy by blocking this pathway.
The expression of AT1mRNA in renal tissue of 4IgA nephropathy rats is positively correlated with the synthesis of collagen type IV, suggesting that activation of RAS system can promote extracellular matrix synthesis. Yiqi Yangyin herbs can significantly inhibit this effect and delay the progression of disease.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692.3

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 肖娟;冬腎丸治療IgA腎病氣陰兩虛證的臨床療效觀察[D];湖南中醫(yī)藥大學(xué);2016年

本文編號(hào)：1545493