本文選題：重癥胰腺炎 + 急性肺損傷　； 參考：《遵義醫(yī)學院》2017年碩士論文

【摘要】：目的:建立重癥急性胰腺炎大鼠模型,觀察重癥急性胰腺炎導致肺損傷過程中NALP3炎性體和水通道蛋白AQP-5的表達以及應用大黃附子湯對SAP-ALI大鼠進行干預后的變化,闡明AQP-5、IL-1β、NALP3在SAP-ALI發(fā)病中的作用。方法:SPF級雄性SD大鼠(購自大連醫(yī)科大學實驗動物中心)24只,體重220-280 g,隨機分為4組:空白對照組(n=6),SAP組(n=6),大黃附子湯治療A組(n=6),大黃附子湯治療B組(n=6)。采用異氟烷麻醉;空白對照組開腹后,找到胰腺,翻動胰腺數(shù)次后關(guān)腹;SAP組、治療組經(jīng)胰膽管緩慢注入4%�；悄懰徕c(1ml/kg)誘導SAP-ALI模型,造模成功后SAP組用2ml生理鹽水于術(shù)后12、24小時灌腸;治療A組用2ml大黃附子湯于術(shù)后12小時灌腸,治療B組用2ml大黃附子湯于術(shù)后12、24h灌腸干預治療。各組大鼠在各自時間點腹主動脈取血測血漿內(nèi)毒素、血淀粉酶、血鈣,在顯微鏡下觀察肺和胰腺組織形態(tài)學變化,通過Smith評分法對實驗各組肺組織進行病理評分,改良Schmidt評分標準對胰腺組織進行病理評分,用ELISA法檢測血清、肺組織IL-1β水平,免疫組化法檢測肺組織AQP-5蛋白表達與分布,Western-blot和免疫組化法檢測肺組織NALP3蛋白表達及分布。結(jié)果:1.血清內(nèi)毒素含量:與空白對照組比較,SAP組血清內(nèi)毒素較對照組明顯升高(P0.05);與SAP組比較,治療組血漿內(nèi)毒素含量較模型組減少(P0.05)。治療B組內(nèi)毒素含量較治療A組降低,差異有統(tǒng)計學意義(P0.05)。2.血清淀粉酶、血鈣含量:與空白對照組比較,SAP組淀粉酶明顯升高,血鈣含量下降(P0.05);與SAP組比較,治療組血淀粉酶含量下降,血鈣含量升高(P0.05)。治療B組淀粉酶較治療A組低,血鈣升高,差異有統(tǒng)計學意義(P0.05)。3.HE染色觀察:空白對照組胰腺腺泡完整;SAP組胰腺壞死明顯,腺體結(jié)構(gòu)紊亂甚至消失,大量炎性細胞浸潤伴出血;治療A組較SAP組腺體水腫減輕,組織間隙寬,炎性細胞浸潤減少,部分有出血和壞死;治療B組腺體輕度水腫,組織間隙輕度增寬,少量炎性細胞浸潤�？瞻讓φ战M肺結(jié)構(gòu)組織正常;SAP組肺泡間隔明顯增寬,肺泡結(jié)構(gòu)破壞,肺泡腔內(nèi)有明顯的出血滲出,大量中性粒細胞浸潤;治療A組肺泡結(jié)構(gòu)有破壞,肺泡間隔增寬較SAP組減輕,肺泡出血及中性粒細胞浸潤減少;治療B組肺泡結(jié)構(gòu)破壞不明顯,肺泡間隔略增寬,中性粒細胞浸潤明顯減少。損傷評分有統(tǒng)計學差異(P0.05)。4.ELLSA:SAP組、治療組血清、肺組織IL-1β與空白對照組比較含量升高(P0.05);治療組與SAP組比較,治療組IL-1β含量較模型組低(P0.05)。治療B組較治療A組比較含量降低(P0.05)。5.免疫組化:與空白對照組比較,SAP組大鼠NALP3高表達,AQP5低表達(P0.05);與SAP組比較,治療組大鼠NALP3表達降低,AQP5表達升高(P0.05)。6.Western-blot:與空白對照組比較,SAP組大鼠NALP3表達量高(P0.05);與SAP組比較,治療組大鼠NALP3表達量降低(P0.05)。結(jié)論:(1)重癥急性胰腺炎肺損傷形成過程中,NALP3表達增加,促使IL-1β的表達增加,此過程降低了AQP-5的表達,使肺內(nèi)水轉(zhuǎn)運出現(xiàn)障礙,肺含水量增多。因此,重癥急性胰腺炎急性肺損傷的發(fā)生可能與NALP3炎性體和AQP-5有關(guān)。(2)大黃附子湯可能通過下調(diào)NALP3炎性小體,促使IL-1β的表達下降,上調(diào)AQP-5表達,從而減輕重癥急性胰腺炎肺損傷程度。
[Abstract]:Objective: to establish a rat model of severe acute pancreatitis (SAP), to observe the expression of NALP3 and aquaporin AQP-5 in the process of severe acute pancreatitis and the changes in the application of Rhubarb Decoction on SAP-ALI rats, and to clarify the role of AQP-5, IL-1 beta, NALP3 in the pathogenesis of SAP-ALI. Methods: SPF grade male SD rats (purchased from the rats). 24 rats at the experimental animal center of Dalian Medical University, weighing 220-280 g, were randomly divided into 4 groups: blank control group (n=6), group SAP (n=6), A group (n=6) treated with Rhubarb Decoction (n=6), rhubarb decoction for B group (n=6) and isoflurane anesthesia. After laparotomy in the blank control group, the pancreas was found, the pancreas was turned over several times, and the group of SAP, the treatment group was injected slowly through the pancreatic bile duct. 4% sodium taurocholate (1ml/kg) was used to induce the SAP-ALI model. After the model was successful, the SAP group was treated with 2ml saline after 12,24 hour postoperatively, and the A group was treated with 2ml Rhubarb decoction for 12 hours after the operation, and the group B was treated with 2ml Rhubarb decoction for 12,24h enema after operation. The blood plasma endotoxin was measured at the abdominal aorta at each time point in each group. The changes of lung and pancreas histomorphology were observed under the microscope by blood amylase and blood calcium. The pathological scores of lung tissues in the experimental groups were evaluated by Smith scoring method. The pathological score of the pancreas tissue was evaluated by improved Schmidt score standard. The serum level of the lung and the level of IL-1 beta in the lung tissue were detected by ELISA, and the expression and distribution of AQP-5 protein in lung tissue was detected by immunohistochemical method. The expression and distribution of NALP3 protein in lung tissue were detected by Western-blot and immunohistochemistry. Results: 1. serum endotoxin content: compared with the blank control group, the serum endotoxin in SAP group was significantly higher than that in the control group (P0.05). Compared with the SAP group, the plasma endotoxin content was lower than that in the model group (P0.05). The content of endotoxin in the treatment group was lower than that in the treatment group of the treatment group, compared with that of the treatment group of the treatment group A. The difference was statistically significant (P0.05).2. serum amylase, blood calcium content: compared with the blank control group, the amylase in the SAP group increased significantly and the blood calcium content decreased (P0.05). Compared with the SAP group, the blood amylase content decreased and the blood calcium content increased (P0.05). In the B group, the amylase was lower than that in the treatment group and the blood calcium increased, the difference was statistically significant (P0.05). 3.HE staining showed that the pancreatic acinus in the blank control group was complete, and the pancreatic necrosis was obvious in group SAP, the structure of the gland was disorganized and even disappeared, and a large number of inflammatory cells were infiltrated with bleeding. In the group A, the edema of the glands in the group SAP was relieved, the space of the tissue was wide, the infiltration of inflammatory cells decreased, and the necrosis of the inflammatory cells, and the mild edema of the glands in the B group and the light widening of the space of the tissue were treated in the B group. The pulmonary structure and tissue in the blank control group were normal, the alveolar space in the SAP group was obviously widened, the alveolar structure was destroyed, the alveolus had obvious bleeding and exudation, and a large number of neutrophils were infiltrated. The alveolar structure in the A group was destroyed, the alveolar septum widened more than that of the SAP group, and the alveolar hemorrhage and neutrophils infiltration were reduced, and the lung of the B group was treated. The damage of the alveolar structure was not obvious, the alveolar septum increased slightly and the neutrophils infiltration decreased significantly. The damage score was statistically different (P0.05).4.ELLSA:SAP, the serum of the treatment group, the IL-1 beta in the lung tissue and the blank control group increased (P0.05); the treatment group was compared with the SAP group, the IL-1 beta content was lower than that in the model group (P0.05). The treatment group was compared with the treatment A for A. Compared with the blank control group, the NALP3 high expression and low expression of AQP5 (P0.05) were compared with the blank control group. Compared with the SAP group, the expression of NALP3 in the treatment group was lower and the AQP5 expression was higher (P0.05).6.Western-blot: than that in the blank control group. The expression of NALP3 in the SAP group was higher than that in the control group. Compared with the SAP group, the expression of the rats was higher than that in the control group. Compared with the SAP group, the expression of NALP3 in the rats was higher than that in the group of SAP. ALP3 expression decreased (P0.05). Conclusion: (1) during the formation of severe acute pancreatitis, the expression of NALP3 is increased and the expression of IL-1 beta is increased. This process reduces the expression of AQP-5, causes the transport of water in the lungs and increases the water content of the lungs. Therefore, the occurrence of acute lung injury in severe acute pancreatitis may be associated with the NALP3 inflammatory and AQP-5. (2) Rhubarb decoction may reduce the expression of IL-1 beta by down regulation of NALP3 inflammatory corpuscle, and increase the expression of AQP-5, thus reducing the degree of lung injury in severe acute pancreatitis.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R576

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