本文選題：硫化氫 + 尿源性膿毒血癥��； 參考：《南華大學(xué)》2014年碩士論文

【摘要】：目的 建立尿源性膿毒血癥兔模型；通過應(yīng)用硫化氫干預(yù),比較兔腎組織細胞因子的表達，明確外源性硫化氫對膿毒血癥兔模型腎組織細胞因子的影響，從而揭示硫化氫在尿源性膿毒血癥腎組織中發(fā)揮作用的可能機制。 方法 1、家兔左側(cè)輸尿管內(nèi)注入大腸桿菌（ATCC25922），并結(jié)扎左側(cè)輸尿管遠端，模擬上尿路急性梗阻并感染所致膿毒血癥兔模型。 2、將48只健康雄性家兔隨機分成4組，每組12只。正常組（Control組）；假手術(shù)組（Sham組）；膿毒血癥組（Sepsis組）；硫氫化鈉組（NaHS組）。 3、每組動物按時間點又分為術(shù)后12小時、術(shù)后24小時、術(shù)后36小時、術(shù)后48小時4個時間點。分別于每個時間點觀測肛溫（RT）、呼吸（RR）、心率（HR）。 4、術(shù)前、術(shù)后12小時、術(shù)后24小時、術(shù)后36小時、術(shù)后48小時分別采血行外周血細胞計數(shù)分析（WBC、中性粒細胞）、腎功能（Cr、BUN）測定。 5、術(shù)后12小時、24小時、36小時、48小時各組家兔腎組織分別行HE染色，光學(xué)顯微鏡下觀察形態(tài)結(jié)構(gòu)變化。Western blot法檢測各組腎組織TNF-α、IL-10的蛋白表達。RT-PCR（reverse translation-PCR）法檢測NF-κB的基因表達。 結(jié)果 1、一般情況：sham組的體溫、心率、呼吸頻率在各個時間點上與control組比較，變化不大（P0.05）；隨造模時間的延長，sepsis組明顯高于control、sham組（P0.05），并呈上升趨勢，NaHS組略低于sepsis組（P0.05）。 2、血液檢測指標(biāo)：各組術(shù)前的血細胞計數(shù)（WBC、中性粒細胞）、Cr、Bun比較，差異無統(tǒng)計學(xué)意義。control組的血細胞計數(shù)（WBC、中性粒細胞）、Cr、Bun在12h、24h、36h、48h與sham組比較，差異無統(tǒng)計學(xué)意義（P0.05）；隨著造模時間的延長，sepsis組明顯高于control、sham組（P0.05），并呈上升趨勢，NaHS組與sepsis組比較，有下降趨勢，其差異有統(tǒng)計學(xué)意義（P0.05）。 3、病理形態(tài)學(xué)：HE染色光鏡下可見sepsis組腎組織大量炎癥細胞浸潤，，腎小球萎縮、腎小囊擴張，腎小管管腔變大。NaHS組較sepsis組上述變化有所減輕。 4、Western Blot法檢測腎組織TNF-α、IL-10的蛋白表達：Control組腎組織TNF-α的蛋白表達和sham組相比較，差異無統(tǒng)計學(xué)意義（P0.05）；sepsis組明顯高于control、sham組，隨造模時間延長，呈上升趨勢（P0.05）；NaHS組與sepsis組比較，有下降趨勢（P0.05）。 Control組腎組織IL-10的蛋白表達和sham組相比較，差異無統(tǒng)計學(xué)意義（P0.05）；sepsis組明顯高于control、sham組，隨造模時間延長，呈上升趨勢（P0.05）；NaHS組與sepsis組比較，仍呈現(xiàn)上升趨勢（P0.05）。 5、RT-PCR（reverse translation-PCR）法檢測NF-κB的基因表達：Control組NF-κB的基因表達和sham組相比較，差異無統(tǒng)計學(xué)意義（P0.05）；sepsis組高于control、sham組，隨造模時間延長，呈現(xiàn)上升趨勢（P0.05）；NaHS組與sepsis組比較，有下降趨勢（P0.05）。 結(jié)論 1、成功建立尿源性膿毒血癥兔模型。 2、外源性硫化氫抑制尿源性膿毒血癥模型腎組織NF-κB及TNF-α的表達，上調(diào)IL-10來減輕尿源性膿毒血癥腎損傷。
[Abstract]:Objective to establish a rabbit model of urine-derived sepsis, compare the expression of cytokines in renal tissues of rabbits with hydrogen sulfide, and clarify the effect of exogenous hydrogen sulfide on renal cytokines in rabbits with sepsis. Methods 1. Escherichia coli ATCC 25922 was injected into the left ureter of rabbits and the distal end of the left ureter was ligated. The rabbit model of sepsis caused by acute upper urinary tract obstruction and infection was simulated. 2 48 healthy male rabbits were randomly divided into 4 groups with 12 rabbits in each group. Control group in normal group; Sham group in sham group; sepsis group in sepsis group; sodium sulfide group in NaHS group. Each group was divided into 4 time points according to the time points: 12 hours after operation, 24 hours after operation, 36 hours after operation and 48 hours after operation. At each time point, we observed the anal temperature, RRV, HR4, 12 hours before operation, 24 hours after operation, 36 hours after operation, and at each time point, we observed RRV, RRV and HR4 before operation, 24 hours after operation and 36 hours after operation, respectively. Blood samples were collected 48 hours after operation for the analysis of WBC, neutrophils and renal function CrBUN.5. the renal tissues of each group were stained by HE staining after 12 hours, 24 hours, 36 hours and 48 hours after operation, respectively. The expression of TNF- 偽 IL-10 in renal tissue was detected by optical microscope. Western blot method was used to detect the expression of NF- 魏 B. results 1. The body temperature, heart rate and respiratory rate of the control group were compared with those of the control group at different time points. With the prolongation of the modeling time, the blood count in sepsis group was significantly higher than that in control sham group, and the increase trend was slightly lower in NaHS group than that in sepsis group. The blood cell count and neutrophils were compared in each group before operation. There was no significant difference in blood cell count between control group and control group. There was no significant difference between control group and sham group at 12 h, 24 h, 36 h or 48 h, but with the prolongation of modeling time, the blood cell count in control group was significantly higher than that in control sham group (P 0.05), and there was an increasing trend between NaHS group and sepsis group. There was a downward trend, and the difference was statistically significant (P 0.05). The pathological morphology showed that there were a large number of inflammatory cells infiltrating, glomerular atrophy, and renal microcapsule dilatation in sepsis group under microscope. Compared with sham group, the protein expression of TNF- 偽 IL-10 in renal tissue was detected by Western blot. There was no significant difference between control group and sham group, and the expression of TNF- 偽 protein in control group was significantly higher than that in control group, and with the prolongation of modeling time, the expression of TNF- 偽 protein in renal tissue of the control group was significantly higher than that in the control group, and the protein expression of TNF- 偽 IL-10 protein in the renal tissue of the control group was significantly higher than that of the control group. Compared with sepsis group, the expression of IL-10 protein in renal tissue of the control group was significantly higher than that of the control group compared with the control group. The expression of IL-10 protein in the control group was significantly higher than that in the controlsham group. With the prolongation of the modeling time, the expression of IL-10 protein in the renal tissue of the control group was significantly higher than that in the control group, and there was an increasing trend between the P0.05NaHS group and the sepsis group. The gene expression of NF- 魏 B was detected by RT-PCR reverse translation (PCR- PCR- PCR) method. There was no significant difference between the control group and the sham group. There was no significant difference between the control group and the control group. With the prolongation of the modeling time, the expression of NF- 魏 B gene in the control group was higher than that in the control group. With the prolongation of the modeling time, the expression of NF- 魏 B gene in the nahs group was higher than that in the sepsis group, and the expression of NF- 魏 B gene in the control group was higher than that in the control group. Conclusion 1. The rabbit model of urine-derived sepsis was successfully established. 2. Exogenous hydrogen sulfide inhibited the expression of NF- 魏 B and TNF- 偽 in renal tissue of the model of urogenic sepsis, and upregulated IL-10 to alleviate renal injury of urine-derived sepsis.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R459.7

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