本文選題：缺血再灌注損傷　切入點(diǎn)：炎癥　出處：《華中科技大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：目的探討地塞米松對小鼠腎臟缺血再灌注損傷的作用及其可能的信號通路。 方法采用8周齡C57BL/6小鼠,隨機(jī)分為3組：(1)假手術(shù)組,假手術(shù)前1小時(shí)腹腔注射生理鹽水；(2)模型組,建立小鼠腎臟缺血再灌注模型,術(shù)前1小時(shí)腹腔注射生理鹽水；(3)實(shí)驗(yàn)組,建立小鼠腎臟缺血再灌注模型,術(shù)前1小時(shí)腹腔注射4mg/kg地塞米松。術(shù)后24小時(shí),收集各組小鼠的血清標(biāo)本和腎臟標(biāo)本,分別檢測血清中肌酐、尿素氮水平,PAS染色觀察腎臟病理改變,免疫組化觀察腎臟中髓過氧化物酶(MPO)的表達(dá),實(shí)時(shí)定量PCR測定腎臟中KIM-1、 TNF-α、IL-6和IFN-γ的轉(zhuǎn)錄水平,Western-blot檢測糖皮質(zhì)激素受體總量和磷酸化程度,P13K p85亞基總量及磷酸化程度,以及AKT總量與磷酸化程度。 結(jié)果假手術(shù)組小鼠的血肌酐和血尿素氮水平分別為(17±5)μmol/L和(15±1.4)mmol/L;模型組分別為(147±13)μmol/L和(70±4)mmol/L:實(shí)驗(yàn)組分別為：(60±7.6)μmol/L和(37±3)mmol/L。假手術(shù)組小鼠腎功能正常,模型組小鼠腎功能下降明顯,實(shí)驗(yàn)組腎功能較模型組有明顯好轉(zhuǎn)(P0.05)。實(shí)驗(yàn)組腎組織KIM-1mRNA水平明顯低于模型組,腎臟損傷程度較輕(P0.05)。腎組織PAS染色示,模型組病變以外髓部位為主,表現(xiàn)為彌漫性腎小管上皮細(xì)胞空泡變性,腎小管上皮多灶狀壞死脫落,并可見大量蛋白質(zhì)管型形成；地塞米松組僅示腎小管上皮細(xì)胞空泡變性,腎小管上皮細(xì)胞局灶狀壞死脫落,未見蛋白管型形成。免疫組化MPO染色,假手術(shù)組沒有MPO陽性細(xì)胞,實(shí)驗(yàn)組MPO陽性細(xì)胞數(shù)明顯少于模型組,實(shí)驗(yàn)組腎組織的MPO陽性細(xì)胞浸潤較模型組減輕(P0.05)。模型組與假手術(shù)組相比,TNF-α、IL-6及IFN-γmRNA水平顯著增高,實(shí)驗(yàn)組三種細(xì)胞因子mRNA水平也有所增加,但明顯低于模型組(P0.05)。糖皮質(zhì)激素受體,PI3K p85亞基和AKT的總量在三組間沒有差異。磷酸化的糖皮質(zhì)激素受體在模型組和實(shí)驗(yàn)組均有明顯升高,但實(shí)驗(yàn)組升高程度更顯著(P0.05)。模型組p85亞基和AKT磷酸化程度明顯增高,實(shí)驗(yàn)組較之顯著下降(P0.05)。 結(jié)論地塞米松通過糖皮質(zhì)激素受體減輕腎臟缺血再灌注中的炎癥反應(yīng),緩解腎臟損傷。PI3K/AKT信號通路可能參與這一作用。
[Abstract]:Objective to investigate the effect of dexamethasone on renal ischemia reperfusion injury in mice and its possible signal pathway. Methods eight week-old C57BL / 6 mice were randomly divided into 3 groups: sham operation group (n = 3). The model group was treated by intraperitoneal injection of normal saline 1 hour before sham-operation. The model of renal ischemia-reperfusion was established. The experimental group was treated with intraperitoneal injection of normal saline for 1 hour before sham-operation. The model of renal ischemia-reperfusion was established in mice. 4 mg / kg dexamethasone was injected intraperitoneally 1 hour before operation. 24 hours after operation, serum samples and kidney samples of each group were collected and serum creatinine was detected. The expressions of myeloperoxidase (MPO) and myeloperoxidase (MPO) in kidney were observed by using urea nitrogen and pas staining. The transcription levels of KIM-1, TNF- 偽, IL-6 and IFN- 緯 in kidney were measured by real-time quantitative PCR. The total amount and phosphorylation of glucocorticoid receptor and phosphorylation of P13K p85 subunit and the total amount and phosphorylation of AKT were detected by Western-blot. Results the levels of serum creatinine and blood urea nitrogen in sham operation group were 17 鹵5 渭 mol/L and 15 鹵1.4 mmol / L, respectively, and those in model group were #number0# 鹵13 渭 mol/L and 70 鹵4 mmol / L respectively: the experimental group was 60 鹵7.6 渭 mol/L and 37 鹵3 mmol / L 路L ~ (-1) respectively. Compared with the model group, the renal function in the experimental group was significantly improved (P 0.05), the level of KIM-1mRNA in the experimental group was significantly lower than that in the model group, and the degree of renal injury was lighter than that in the model group. The renal tissue PAS staining showed that the lesion was mainly located outside the medulla in the model group. Diffuse tubular epithelial cell vacuolation, multiple focal necrosis and exfoliation of renal tubular epithelium, and formation of a large number of protein tubules were observed in the dexamethasone group, while in the dexamethasone group, the vacuolar degeneration of the tubular epithelial cells was observed only in the dexamethasone group. There were no MPO positive cells in the sham-operated group, and the number of MPO positive cells in the experimental group was significantly lower than that in the model group, and the number of MPO positive cells in the experimental group was significantly lower than that in the model group. The infiltration of MPO positive cells in the experimental group was less than that in the model group (P 0.05). The levels of TNF- 偽 IL-6 and IFN- 緯 mRNA in the model group were significantly higher than those in the sham operation group, and the mRNA levels of the three cytokines in the experimental group were also increased. The total amount of PI3K p85 subunit and AKT in the model group was significantly lower than that in the model group. The phosphorylated glucocorticoid receptor was significantly increased in both the model group and the experimental group. The phosphorylation of p85 subunit and AKT in the model group was significantly higher than that in the experimental group, but the level of P0.05 in the experimental group was significantly lower than that in the control group. Conclusion Dexamethasone can attenuate the inflammatory reaction in renal ischemia reperfusion by glucocorticoid receptor and attenuate renal injury. PI3K / AKT signaling pathway may be involved in this role.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R965

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李春艷,成小松;急性腎功能衰竭動物模型的研究進(jìn)展[J];中國急救醫(yī)學(xué);2002年08期

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本文編號：1632787