本文選題：置管 + 單側(cè)輸尿管梗阻再通��； 參考：《南京大學(xué)》2017年碩士論文

【摘要】：目的:用小鼠作為模型動(dòng)物,探索改良現(xiàn)有的單側(cè)輸尿管梗阻再通(RUUO)模型的方法;在此基礎(chǔ)上,研究氯沙坦對(duì)解除梗阻后的腎臟的纖維化的影響,以及調(diào)節(jié)性T細(xì)胞在這一過(guò)程中的變化。方法:將18只C57BL/6小鼠隨機(jī)分為RUUO+氯沙坦組(n=6)、RUUO組(n=6)和假手術(shù)組(n=6)。在左側(cè)輸尿管梗阻(UUO)7天后采用套入+置管法實(shí)施再通手術(shù)(RUUO)。再通術(shù)后RUUO+氯沙坦組水飼法給予氯沙坦。UUO第5天、RUUO第4、7天行外周血流式細(xì)胞術(shù)檢測(cè)調(diào)節(jié)性T細(xì)胞(Treg);RUUO術(shù)后7天,取術(shù)側(cè)腎組織行HE、Masson染色,同時(shí)免疫組化檢測(cè)腎臟α-SMA表達(dá),用Western blot檢測(cè)腎臟Foxp3蛋白的表達(dá)。結(jié)果:RUUO+氯沙坦組和RUUO組腎臟均無(wú)明顯積水,體積較對(duì)側(cè)縮小,色澤較淺,腎盂無(wú)明顯積水,提示梗阻解除,尿路再通成功。HE染色結(jié)果示RUUO組腎小管上皮紊亂,刷狀緣脫落,管腔擴(kuò)張,管型形成;假手術(shù)組腎小管結(jié)構(gòu)清晰,上皮完整;RUUO+氯沙坦組病理改變較手術(shù)組為輕。RUUO組腎小管損傷評(píng)分(3.9±0.74)較RUUO+氯沙坦組(2.4±0.48)為重(p0.05),假手術(shù)組腎小管損傷評(píng)分(0.5±0.45)較兩者輕(p0.05)。Masson染色結(jié)果示假手術(shù)組未見(jiàn)明顯藍(lán)染區(qū)域,RUUO組可見(jiàn)明顯藍(lán)染區(qū)域,主要分布在腎小管間質(zhì)中,而RUUO+氯沙坦組較RUUO組藍(lán)染面積減少。RUUO+氯沙坦組、RUUO組和假手術(shù)組膠原面積所占比例分別是5.33±0.64%、9.60± 1.49%和1.17±0.41%,各組之間存在統(tǒng)計(jì)學(xué)區(qū)別(p0.05)。腎臟α-SMA免疫組化染色示假手術(shù)組α-SMA主要表達(dá)于小血管管壁,亦散見(jiàn)于腎小球周圍和腎小管間質(zhì)中,表達(dá)量較少。RUUO組α-SMA表達(dá)量明顯增多,主要見(jiàn)于腎小管間質(zhì)中。RUUO+氯沙坦組的表達(dá)量較RUUO組減少。RUUO+氯沙坦組α-SMA免疫組化的平均積分光密度為0.0138±0.0011,較RUUO組(0.0203±0.0012)小(P0.05),假手術(shù)組α-SMA的平均積分光密度為0.0038±0.0003,與其余兩組相比有統(tǒng)計(jì)學(xué)差異(P0.05)。單側(cè)輸尿管結(jié)扎術(shù)(UUO)后第5天、再通手術(shù)(RUUO)后第4天以及第7天外周血Foxp3+Treg/CD4+T cell比例示RUUO+氯沙坦組、RUUO組和假手術(shù)組之間無(wú)統(tǒng)計(jì)學(xué)區(qū)別。Western blot結(jié)果示腎臟Foxp3蛋白在RUUO組表達(dá)量降低,其積分光密度與內(nèi)參積分光密度之比為 0.0664±0.0080,分別低于 RUUO+氯沙坦組的 0.5987±0.2543(P0.05)和假手術(shù)組的0.9910±0.1484(P0.05);RUUO+氯沙坦組Foxp3蛋白的表達(dá)量低于假手術(shù)組(P0.05)。結(jié)論:套入+置管法可以建立起較為可靠的小鼠單側(cè)輸尿管梗阻再通模型;單側(cè)輸尿管梗阻再通后腎臟仍然處于損傷和纖維化狀態(tài);調(diào)節(jié)性T細(xì)胞的標(biāo)志物Foxp3在RUUO腎臟中的表達(dá)量下調(diào);氯沙坦可以提高RUUO腎臟Foxp3蛋白的表達(dá)量并且改善單側(cè)輸尿管梗阻再通后腎臟的纖維化指標(biāo)。
[Abstract]:Objective: to study the effect of losartan on renal fibrosis after unilateral ureteral obstruction (UUO) was removed by using mice as model animals. And the changes of regulatory T cells during this process. Methods: eighteen C57BL / 6 mice were randomly divided into RUUO losartan group and sham operation group. After 7 days of UUO obstruction in left ureter recanalization was performed by intubation. Losartan 路UUO group was treated with losartan. UUO on the 5th day and RUUO on the 4th and 7th day after operation, the regulatory T cell Tregon was detected by peripheral blood flow cytometry 7 days after operation. The renal tissues were taken out for HE Masson staining, and the expression of 偽 -SMA in the kidney was detected by immunohistochemistry, and the expression of 偽 -SMA in the kidney was also detected by immunohistochemistry, and the expression of 偽 -SMA in the kidney was detected by immunohistochemistry. The expression of Foxp3 protein in kidney was detected by Western blot. Results there was no obvious hydronephrosis in the kidney of the two groups, the volume was smaller than the opposite side, the color of the renal pelvis was lighter, and there was no obvious hydronephrosis in the renal pelvis. The results indicated that the obstruction was relieved and the urethral recanalization was successful. The results of HE staining showed that the renal tubule epithelium in the RUUO group was disordered and the brush edge was shedding. The renal tubules in the sham-operation group were clear, and the tubules were dilated and formed. The pathological changes in the epithelial integrity group were lighter than those in the operation group (3.9 鹵0.74) and the renal tubule injury score in the sham-operated group (0.5 鹵0.45) was significantly higher than that in the RUUO losartan group (2.4 鹵0.48). The results of Masson staining showed that there was no significant blue in the sham-operated group. The dyed area can be seen in the Ruud group, and the blue staining area can be seen. The percentage of collagen in RUUO losartan group and sham operation group was 5.33 鹵0.64 鹵1.49% and 1.17 鹵0.41%, respectively. Renal 偽 -SMA immunohistochemical staining showed that 偽 -SMA was mainly expressed in the wall of small vessels, scattered around glomeruli and tubulointerstitium in sham-operated group, and the expression of 偽 -SMA was significantly increased in RUUO group. The average integral optical density of 偽 -SMA immunohistochemistry in the RUUO losartan group was 0.0138 鹵0.0011, which was higher than that in the RUUO group (0.0203 鹵0.0012). The mean integral optical density of 偽 -SMA in the sham-operated group was 0.0038 鹵0.0003, and that in the other two groups was 0.0038 鹵0.0003. There was a statistical difference between the two groups (P 0.05). The ratio of Foxp3 Treg-CD4T cell in peripheral blood on the 4th and 7th day after UUO (unilateral ureteral ligation) showed no statistical difference between RUUO Losartan group and sham-operated group. Western blot results showed that the expression of Foxp3 protein in kidney decreased in RUUO group. The ratio of integral optical density to integrated optical density of internal parameters was 0.0664 鹵0.0080, which was lower than that of Rup losartan group (0.5987 鹵0.2543 P0.05) and sham-operated group (0.9910 鹵0.1484 P0.05P 0.05). The expression of Foxp3 protein in Ruluo losartan group was lower than that in sham operation group (P0.05. Conclusion: the recanalization model of unilateral ureteral obstruction in mice can be established by inserting catheter, and the kidney is still in the state of injury and fibrosis after the recanalization of unilateral ureteral obstruction. The expression of Foxp3, a marker of regulatory T cells, was down-regulated in RUUO kidney, and losartan could increase the expression of Foxp3 protein and improve the renal fibrosis index after the recanalization of unilateral ureteral obstruction.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R692
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本文編號(hào)：2041669