【摘要】：目的通過建立大鼠重癥急性胰腺炎(severe acute pancreatitis,SAP)動(dòng)物模型,研究趨化因子受體CX3CR1在SAP大鼠模型胰腺組織及相關(guān)受累器官組織中的表達(dá),探討其能否作為SAP發(fā)生的預(yù)測(cè)指標(biāo)。方法60只健康成年雄性Sprague-Dawley大鼠,體重200-250g,隨機(jī)分為假手術(shù)(Sham operation,SO)組(n=20)、輕型急性胰腺炎(mild acute pancreatitis,MAP)組(n=20)及SAP組(n=20),分別以0.5%及5%�；悄懰徕c建立MAP及SAP動(dòng)物模型,于制模后6 h、12 h、24 h及48 h處死大鼠。光鏡下觀察各組大鼠胰腺組織病理學(xué)表現(xiàn);采用速率法檢測(cè)各組大鼠血清淀粉酶水平;應(yīng)用酶聯(lián)免疫吸附試驗(yàn)(Enzyme-linked immunosorbent assay,ELISA)法檢測(cè)各組大鼠血清中CX3CR1及TNF-α的變化;同時(shí)應(yīng)用蛋白質(zhì)印跡(Western blot)法及免疫組織化學(xué)法檢測(cè)各組大鼠胰腺、肺臟及腎臟組織中CX3CR1蛋白的表達(dá);分析血清CX3CR1與大鼠胰腺病理?yè)p傷評(píng)分及血清TNF-α水平之間的相關(guān)性。結(jié)果1.SAP組各時(shí)間點(diǎn)(6h、12h、24h、48h)大鼠胰腺組織病理學(xué)評(píng)分均較SO組及MAP組相應(yīng)時(shí)間點(diǎn)顯著升高(P0.05)。MAP組各時(shí)間點(diǎn)大鼠胰腺組織病理學(xué)評(píng)分較SO組各相應(yīng)時(shí)間點(diǎn)顯著升高(P0.05)。2.SAP組各時(shí)間點(diǎn)大鼠血清淀粉酶水平較MAP組及SO組相應(yīng)時(shí)間點(diǎn)均顯著升高(P0.05)。3.SAP組大鼠血清CX3CR1及TNF-α水平均自制模后逐漸升高,24h達(dá)高峰,之后逐漸下降。與MAP組及SO組各相應(yīng)時(shí)間點(diǎn)比較,均顯著升高(P0.05)。4.Western blot法結(jié)果顯示:SAP組大鼠胰腺及肺臟組織CX3CR1蛋白表達(dá)水平自制模后逐漸升高,24h達(dá)高峰(2.19±0.26,2.24±0.26),48h(1.89±0.23,2.02±0.27)后逐漸下降。SAP組腎臟組織CX3CR1蛋白表達(dá)水平自制模后逐漸升高,48h達(dá)高峰(2.67±0.21),與MAP組及SO組相應(yīng)時(shí)間點(diǎn)比較均顯著升高(P0.05)。5.免疫組織化學(xué)法結(jié)果顯示:SAP組大鼠胰腺、肺臟及腎臟組織均有CX3CR1表達(dá),且表達(dá)水平顯著高于MAP組及SO組,其中胰腺及肺臟組織中CX3CR1表達(dá)在制模后24h達(dá)高峰,腎臟組織中CX3CR1蛋白表達(dá)在制模后48h達(dá)高峰。6.相關(guān)性分析結(jié)果:Spearman等級(jí)相關(guān)分析表明SAP組血清CX3CR1水平與胰腺病理?yè)p傷評(píng)分呈正相關(guān)(r=0.711,P0.01);Pearson相關(guān)分析表明SAP組血清CX3CR1水平與血清TNF-α水平呈正相關(guān)(r=0.828,P0.01)。結(jié)論1.趨化因子受體CX3CR1參與大鼠SAP發(fā)生發(fā)展過程,且其血清水平與胰腺組織病理?yè)p傷嚴(yán)重程度有關(guān),可能能作為預(yù)測(cè)急性胰腺炎嚴(yán)重程度的有效指標(biāo)之一,值得進(jìn)一步深入研究。2.CX3CR1可能參與大鼠SAP繼發(fā)急性肺損傷(acute lung injury,ALI)過程。3.CX3CR1可能參與大鼠SAP繼發(fā)急性腎損傷(acute kidney injury,AKI)過程。
[Abstract]:Objective to study the expression of chemokine receptor (CX3CR1) in pancreatic tissues and related involved organs of severe acute pancreatitis (SAP) in rats, and to explore whether it can be used as a predictor for the occurrence of severe acute pancreatitis (SAP). Methods Sixty healthy adult male Sprague-Dawley rats, weighing 200-250 g, were randomly divided into three groups: sham operation group (n = 20), mild acute pancreatitis (map) group (n = 20) and SAP group (n = 20). Map and SAP animal models were established with 0.5% and 5% sodium taurocholate, respectively. The rats were sacrificed at 6 h, 12 h, 24 h and 48 h. The histopathological manifestations of pancreas were observed under light microscope, the serum amylase level was detected by rate method, and the changes of CX3CR1 and TNF- 偽 in serum were detected by Enzyme-linked immunosorbent assay-ELISA (Elisa). Western blot and immunohistochemistry were used to detect the expression of CX3CR1 protein in pancreas, lung and kidney of rats, and the correlation between serum CX3CR1 and pancreatic pathological injury score and serum TNF- 偽 level was analyzed. Results 1. The histopathological scores of pancreas in SAP group were significantly higher than those in so group and map group (P0.05). The histopathological score of SAP group was significantly higher than that of so group at each time point (P0.05). 2. SAP group had significantly higher pancreatic histopathological score than that of so group at each time point (P0.05). The levels of serum amylase in SAP group were significantly higher than those in map group and so group (P0.05). 3. The levels of CX3CR1 and TNF- 偽 in SAP group were increased gradually and reached the peak at 24 h after self-made model. Then it gradually declined. Compared with map group and so group, The expression level of CX3CR1 protein in pancreas and lung tissues of rats in the SAP group was significantly increased (P0.05). 4. The expression level of CX3CR1 protein in the pancreas and lung tissues of the rats in the SAP group gradually increased to a peak of (2.19 鹵0.26) 2.24 鹵0.26 after 24 hours and gradually decreased after 48 hours (1.89 鹵0.23 鹵2.02 鹵0.27). The expression level of CX3CR1 protein in the renal tissue of SAP group gradually increased after 48 hours. The peak value was (2.67 鹵0.21), which was significantly higher than that in map group and so group (P0.05). The expression of CX3CR1 in pancreas, lung and kidney of rats in SAP group was significantly higher than that in map group and so group. The expression of CX3CR1 in pancreas and lung reached its peak 24 hours after modeling. The expression of CX3 CR1 protein reached its peak at 48h after modeling. The correlation analysis showed that the serum CX3CR1 level was positively correlated with the pancreatic pathological injury score in SAP group. Pearson correlation analysis showed that there was a positive correlation between serum CX3CR1 level and serum TNF- 偽 level in SAP group (r = 0.828, P 0.01). Conclusion 1. The chemokine receptor CX3CR1 is involved in the pathogenesis and development of SAP in rats. The serum level of CX3CR1 is related to the severity of pancreatic pathological injury. CX3CR1 may be one of the effective indicators to predict the severity of acute pancreatitis. 2. CX3CR1 may be involved in the process of acute lung injury (acute lung injuryto Ali). 3. CX3CR1 may be involved in the process of acute renal injury (acute kidney injura).
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R576

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