本文選題：新生鼠 + 腎積水��； 參考：《遵義醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：通過(guò)制作腰大肌壓迫輸尿管，建立新生鼠腎積水動(dòng)物模型。應(yīng)用黃芪、冬蟲夏草提取液對(duì)新生鼠腎積水動(dòng)物模型進(jìn)行皮下注射，檢測(cè)動(dòng)物模型中梗阻建立后病腎組織轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）、胰島素樣生長(zhǎng)因子-1（IGF-1）的表達(dá)變化，以及檢測(cè)病腎尿TGF-β1、IGF-1的水平。以探討黃芪、冬蟲夏草對(duì)新生鼠腎積水病腎的損害有否保護(hù)作用，從而為臨床更好地治療先天性腎積水提供一種新的思路。 方法： 1、動(dòng)物模型的建立選用生后48h內(nèi)的Wistar新生鼠96只，隨機(jī)分為8組，每組12只。隨機(jī)選擇1組行假手術(shù)，作為對(duì)照組；余7組采用腰大肌壓迫部分左輸尿管制作新生鼠腎積水動(dòng)物模型，任選其中1組作為模型組，余各組再隨機(jī)分為干預(yù)1組~干預(yù)6組： ①、組1：對(duì)照組（假手術(shù)組）：手術(shù)后（術(shù)中暴露左輸尿管后即關(guān)閉手術(shù)切口）Wistar新生鼠皮下注射生理鹽水（1ml/kg/天）。 ②、組2：模型組：手術(shù)后（術(shù)中建立左側(cè)輸尿管部分梗阻模型）Wistar新生鼠皮下注射生理鹽水（1ml/kg/天）。 ③、干預(yù)1組（黃芪組1-低劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射低劑量的黃芪注射液（1ml/kg/天），飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 ④、干預(yù)2組（黃芪組2-高劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射高劑量的黃芪注射液（3ml/kg/天），飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 ⑤、干預(yù)3組（冬蟲夏草組1-低劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射低劑量的冬蟲夏草提取液（0.5ml/kg/天），飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 ⑥、干預(yù)4組（冬蟲夏草組2-高劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射高劑量的冬蟲夏草提取液（2.0ml/kg/天），飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 ⑦、干預(yù)5組（低劑量黃芪+冬蟲夏草組-低劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射低劑量的黃芪注射液（1ml/kg/天）和冬蟲夏草提取液（0.5ml/kg/天），飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 ⑧、干預(yù)6組（高劑量黃芪+冬蟲夏草組-高劑量組）：動(dòng)物模型建立后Wistar新生鼠皮下注射高劑量的黃芪注射液（3ml/kg/天）和冬蟲夏草提取液（2.0ml/kg/天），，飼養(yǎng)3周標(biāo)本取材后、頸椎脫髓法處死動(dòng)物。 2、各種指標(biāo)的檢測(cè)：(1)、采集下列各組：對(duì)照組、模型組、干預(yù)組1-6的病腎組織標(biāo)本，部分-80℃冰箱保存、部分甲醛固定后做成石蠟標(biāo)本待檢：①、石蠟標(biāo)本切片進(jìn)行HE染色后、光鏡下觀察組織結(jié)構(gòu)的變化；②、石蠟標(biāo)本切片后、通過(guò)免疫組織化學(xué)PV法檢測(cè)轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）、胰島素樣生長(zhǎng)因子-1（IGF-1）的表達(dá)，應(yīng)用Image-ProPlus圖像分析軟件測(cè)量平均光密度值；③、采用RT-PCR技術(shù)檢測(cè)病腎組織內(nèi)TGF-β1mRNA、 IGF-1mRNA的表達(dá)變化；(2)、動(dòng)物處死時(shí)收集各組病腎腎盂尿液，應(yīng)用ELISA法檢測(cè)尿TGF-β1、尿IGF-1的水平。 結(jié)果： 1、動(dòng)物模型建立期間，動(dòng)物的存活情況： 新生Wistar鼠共96只，建立腎積水梗阻模型及喂養(yǎng)期間死亡5只（麻醉過(guò)深致死1只、術(shù)中死亡2只、喂養(yǎng)期間死亡2只），存活91只（94.79%）。 2、病腎組織形態(tài)學(xué)的變化： (1)、肉眼觀：對(duì)照組雙腎大小、形態(tài)一致，腎包膜光整，腎實(shí)質(zhì)質(zhì)地等同；模型組病腎明顯增大，腎包膜張力高，腎包膜與腎實(shí)質(zhì)粘連重、不易分離，腎實(shí)質(zhì)菲薄如紙，腎盂明顯擴(kuò)張、積水；干預(yù)組：病腎增大，但小于模型組病腎，腎實(shí)質(zhì)薄，腎盂擴(kuò)張、積水。 （2）、光鏡下病腎組織結(jié)構(gòu)的變化： 對(duì)照組：雙腎腎小球結(jié)構(gòu)、腎小管結(jié)構(gòu)發(fā)育正常，腎小囊未見擴(kuò)張。模型組：病腎腎小球結(jié)構(gòu)形態(tài)怪異、失常，腎小球鮑曼氏囊明顯擴(kuò)張，腎小管明顯擴(kuò)張。干預(yù)組：病腎腎小球形態(tài)結(jié)構(gòu)異常，腎小球鮑曼氏囊輕度擴(kuò)張，腎小管擴(kuò)張，但腎小球、腎小管的損害較模型組明顯減輕，隨藥物劑量的增大、以及聯(lián)合用藥，病腎組織結(jié)構(gòu)的損害呈逐漸減輕的趨勢(shì)。 3、病腎組織內(nèi)TGF-β1、IGF-1蛋白水平與轉(zhuǎn)錄水平的表達(dá) 對(duì)照組腎組織內(nèi)TGF-β1低表達(dá)，TGF-β1mRNA也低表達(dá)；腎組織內(nèi)IGF-1高表達(dá)，IGF-1mRNA也高表達(dá)。模型組病腎組織內(nèi)TGF-β1高表達(dá)，TGF-β1mRNA表達(dá)明顯升高；病腎組織內(nèi)IGF-1低表達(dá)，IGF-1mRNA表達(dá)明顯減低。干預(yù)各組病腎組織內(nèi)TGF-β1均有不同程度的表達(dá)，低劑量組病腎TGF-β1的表達(dá)比高劑量組、聯(lián)合用藥組增多，但仍明顯低于模型組；在病腎組織內(nèi)IGF-1也有不同程度的表達(dá)，但低劑量組病腎IGF-1的表達(dá)比高劑量組、聯(lián)合用藥組減低，仍明顯高于模型組。對(duì)照組與模型組病腎組織中相應(yīng)TGF-β1、IGF-1水平間比較，差異均有統(tǒng)計(jì)學(xué)意義（P0.05），模型組與干預(yù)各組病腎組織中相應(yīng)TGF-β1、IGF-1水平間比較，差異也有統(tǒng)計(jì)學(xué)意義（P0.05），對(duì)照組與干預(yù)各組病腎組織中相應(yīng)TGF-β1、IGF-1水平間比較，差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。干預(yù)各組相應(yīng)的低劑量組與高劑量組病腎組織TGF-β1、IGF-1水平間比較，差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。 4、各組病腎腎盂尿內(nèi)TGF-β1、IGF-1水平的變化 在對(duì)照組中腎盂尿液TGF-β1含量低，而IGF-1含量高。模型組中病腎腎盂尿液中TGF-β1含量高，而IGF-1含量低。干預(yù)各組病腎腎盂內(nèi)尿液TGF-β1、IGF-1水平介于對(duì)照組與模型組之間。對(duì)照組與模型組相應(yīng)病腎尿TGF-β1、IGF-1水平間比較，差異均有統(tǒng)計(jì)學(xué)意義（P0.05）；模型組與干預(yù)各組相應(yīng)病腎尿TGF-β1、IGF-1水平間比較，差異也有統(tǒng)計(jì)學(xué)意義（P0.05）；對(duì)照組與干預(yù)各組相應(yīng)病腎尿TGF-β1、IGF-1水平間比較，差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。干預(yù)各組相應(yīng)的低劑量組與高劑量組病腎尿TGF-β1、IGF-1水平間比較，差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論： 1、應(yīng)用黃芪、冬蟲夏草對(duì)新生鼠腎積水動(dòng)物模型進(jìn)行皮下注射表明：新生鼠病腎組織及尿內(nèi)TGF-β1的表達(dá)較模型組減低，而病腎IGF-1水平較模型組升高。 2、黃芪、冬蟲夏草可能通過(guò)下調(diào)新生鼠腎積水病腎組織內(nèi)TGF-β1的表達(dá)、或/和上調(diào)病腎IGF-1水平，從而對(duì)新生鼠腎積水病腎的損害有一定的保護(hù)作用。
[Abstract]:Objective : To establish an animal model of neonatal rat renal hydrops by making lumbar muscle compression ureter . The expression of TGF - 尾1 ( TGF - 尾1 ) , insulin - like growth factor - 1 ( IGF - 1 ) and the level of TGF - 尾1 and IGF - 1 were detected in animal model .

Method :

1 . 96 Wistar rats were randomly divided into 8 groups randomly divided into 8 groups ( n = 12 ) .
The rats were randomly divided into the intervention group 1 group and the intervention group 6 group .

Group 1 : Control group ( sham operation group ) : normal saline ( 1 ml / kg / day ) was injected subcutaneously into Wistar neonatal rats after operation ( i.e . , the operation incision was closed after the left ureter was exposed ) .

( 2 ) Group 2 : Model group : After operation ( the left ureter partial obstruction model was established ) , the rats were subcutaneously injected with physiological saline ( 1 ml / kg / day ) .

( 3 ) Group 1 ( 1 - low - dose group ) : A low - dose astragalus injection ( 1 ml / kg / day ) was injected subcutaneously into Wistar rats after the animal model was established , and the animals were sacrificed after three weeks of feeding .

4 . Intervention group 2 ( astragalus group 2 - high dose group ) : Wistar rats were subcutaneously injected with high - dose astragalus injection ( 3 ml / kg / day ) after the animal model was established . After 3 weeks of feeding , the animals were sacrificed .

( 5 ) The rats were treated with low - dose Cordyceps extract ( 0.5 ml / kg / day ) by subcutaneous injection of low - dose Cordyceps sinensis ( 0.5 ml / kg / day ) after the animal model was established , and the animals were sacrificed after 3 weeks of feeding .

6 . Intervention 4 groups ( Cordyceps group 2 - high dose group ) : A high - dose Cordyceps extract ( 2.0ml / kg / day ) was subcutaneously injected into Wistar rats after the animal model was established , and the animals were sacrificed after three weeks of feeding .

7 . Intervention 5 groups ( low - dose astragalus + cordyceps group - low - dose group ) : Wistar rats were subcutaneously injected with low - dose astragalus injection ( 1 ml / kg / day ) and Cordyceps extract ( 0.5 ml / kg / day ) after the animal model was established , and the animals were sacrificed after three weeks of feeding .

8 . Intervention 6 groups ( high - dose astragalus + Cordyceps group - high - dose group ) : Wistar rats were subcutaneously injected with high - dose astragalus injection ( 3 ml / kg / day ) and Cordyceps extract ( 2.0ml / kg / day ) after the animal model was established , and the animals were sacrificed after 3 weeks after feeding .

2 . Detection of various indexes : ( 1 ) The following groups were collected : control group , model group , and intervention group 1 - 6 disease kidney tissue specimen , part -80 鈩

本文編號(hào)：1864691