本文選題：腎素-血管緊張素系統(tǒng) + 非肽類腎素抑制劑��； 參考：《河北聯(lián)合大學(xué)》2014年碩士論文

【摘要】：目的腎素-血管緊張素系統(tǒng)（Renin-angiotensin system，RAS）失調(diào)在高血壓及高血壓腎臟損傷的發(fā)生發(fā)展中起重要作用。腎素（renin）是啟動(dòng)RAS第一步的限速酶，對(duì)RAS的活化有至關(guān)重要的作用。本研究通過觀察阿利克侖（Aliskiren）對(duì)人血管緊張素原-腎素（Angiotensinogen-renin，AGT-REN）雙轉(zhuǎn)基因高血壓小鼠血壓、腎損傷指標(biāo)以及循環(huán)和腎臟局部RAS成分表達(dá)的變化，探討非肽類腎素抑制劑阿利克侖對(duì)高血壓及腎臟損傷的影響及其作用機(jī)制。 方法1動(dòng)物分組及給藥方法：12只10月齡雄性AGT-REN雙轉(zhuǎn)基因高血壓小鼠隨機(jī)分為高血壓對(duì)照組（HT組）和阿利克侖治療組（HT+Aliskiren組），每組6只，另選擇背景相同的6只野生型C57B6雄性小鼠作為正常對(duì)照組（WT）。給藥方法：單次給藥采用灌胃經(jīng)口途徑，分別給予5mg/kg、10mg/kg、20mg/kg阿利克侖，監(jiān)測(cè)給藥后24h內(nèi)血壓；持續(xù)給藥通過皮下埋置微量滲透泵，以20mg/kg/d對(duì)HT+Aliskiren組小鼠給予阿利克侖處理28d，其余兩組給予生理鹽水作為空白對(duì)照，期間監(jiān)測(cè)血壓，28d后取材，進(jìn)行各項(xiàng)指標(biāo)檢測(cè)。2動(dòng)脈血壓監(jiān)測(cè)：采用無創(chuàng)尾套法測(cè)量各組小鼠在清醒與安靜狀態(tài)下給藥前后的平均動(dòng)脈壓（MAP），觀察阿利克侖單次給藥后24h內(nèi)血壓以及持續(xù)給藥28d內(nèi)隔日血壓的變化。3腎功能損傷指標(biāo)檢測(cè)：①考馬斯亮藍(lán)（CBB）結(jié)合法測(cè)定24h尿蛋白含量；②全自動(dòng)生化分析儀測(cè)定血清肌酐、尿素；③光鏡（HE、Masson染色）和電鏡觀察腎臟組織病理形態(tài)改變。4氧化應(yīng)激反應(yīng)指標(biāo)檢測(cè)：化學(xué)發(fā)光法測(cè)定腎組織勻漿超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量以及蛋白免疫印跡法（Western blot, WB）檢測(cè)腎組織p47phox蛋白表達(dá)變化。5RAS成分表達(dá)變化檢測(cè)：ELISA法測(cè)定血清和腎組織勻漿AngⅡ和Ang(1-7)含量；WB檢測(cè)腎組織ACE、ACE2蛋白表達(dá)。 結(jié)果1阿利克侖的降壓效果：與WT小鼠比較，HT組小鼠MAP明顯升高；單次和持續(xù)給藥實(shí)驗(yàn)均顯示，HT+Aliskiren組小鼠MAP均較HT組明顯降低。2HT小鼠24h尿蛋白、血清尿素、肌酐均較WT小鼠明顯升高；在HT+Aliskiren組小鼠上述腎功能損傷指標(biāo)較HT組明顯降低。3病理形態(tài)學(xué)結(jié)果顯示：光鏡下HT組小鼠腎臟實(shí)質(zhì)厚度較WT組明顯變薄，HT+Aliskiren組上述病理損傷得到改善。與WT小鼠比較，HT組小鼠腎小球硬化面積評(píng)分和腎間質(zhì)損傷評(píng)分明顯增加，HT+Aliskiren組評(píng)分結(jié)果較HT組降低。電鏡下，HT小鼠腎小球基底膜增厚，足突融合甚至消失，腎小管上皮細(xì)胞胞漿內(nèi)線粒體水腫、空泡變性，間質(zhì)可見大量膠原纖維沉積，；HT+Aliskiren組小鼠腎臟上述病變較HT組明顯減輕。4與WT小鼠比較，HT小鼠腎臟組織SOD活性降低，MDA生成增加，p47phox蛋白表達(dá)明顯增多；與HT組小鼠比較，HT+Aliskiren組小鼠腎臟組織SOD活性增加，MDA的生成減少，p47phox蛋白表達(dá)明顯減少。5與WT組比較，HT組小鼠ACE蛋白表達(dá)增加，ACE2蛋白表達(dá)下降，AngⅡ/Ang(1-7)比值增加；與HT組小鼠相比，阿利克侖可下調(diào)HT小鼠腎組織ACE蛋白表達(dá)，上調(diào)ACE2表達(dá)，降低血漿和腎組織AngⅡ/Ang(1-7)比值。 結(jié)論1阿利克侖具有良好的降壓效果及腎臟保護(hù)作用。2阿利克侖通過抑制AngⅡ的生成，降低腎臟局部AngⅡ/Ang(1-7)比值，維持腎臟局部RAS穩(wěn)態(tài)平衡。3腎素抑制劑從源頭上抑制RAS激活，但對(duì)ACE2-Ang(1-7)無明顯抑制作用，且可上調(diào)腎臟ACE2蛋白表達(dá)，這一作用可能與減少了AngⅡ生成有關(guān)。4阿利克侖可通過抑制NADPH氧化酶表達(dá)，，降低氧化應(yīng)激水平而發(fā)揮腎臟保護(hù)作用。
[Abstract]:Objective Renin - angiotensin system ( RAS ) imbalance plays an important role in the development of hypertension and hypertensive kidney injury . Renin is a limiting enzyme for the first step of RAS , which plays an important role in the activation of RAS .

Method 1 Animal grouping and administration were divided into two groups : hypertension control group ( HT group ) and aliskiren group ( HT + Aliskiren group ) .
The mean arterial pressure ( MAP ) before and after administration of 5 mg / kg / d to HT + Aliskiren group was monitored by subcutaneous implantation of micro osmotic pump . The average arterial pressure ( MAP ) before and after administration of the rats in conscious and quiet state was measured by noninvasive method .
( 2 ) measuring serum creatinine and urea by a full - automatic biochemical analyzer ;
The activity of superoxide dismutase ( SOD ) , the content of malondialdehyde ( MDA ) and the expression of p47phox protein were detected by Western blot and WB .
The expression of ACE and ACE2 protein in renal tissue was detected by WB .

Results Compared with WT mice , MAP in HT group increased significantly compared with WT mice .
Both single and continuous administration showed that the MAP in HT + Aliskiren group was significantly lower than that in HT group . 24h urinary protein , serum urea and creatinine of 2HT mice were significantly higher than those in WT mice .
The results showed that the renal parenchyma thickness of HT group mice was significantly lower than that in WT group and HT + Aliskiren group was significantly higher than that in HT group .
Compared with WT mice , the activity of SOD in renal tissue of HT mice decreased , MDA formation increased , and the expression of p47phox protein increased significantly .
Compared with the HT group , the activity of SOD in kidney of HT + Aliskiren group increased , the expression of MDA was decreased , the expression of p47phox protein decreased significantly . Compared with WT group , the expression of ACE protein in HT group increased , the expression of ACE2 protein decreased , and the ratio of Ang 鈪

本文編號(hào)：1974430