大鼠急危重症胰腺炎并发心脏损害的相关机制研究

发布时间：2018-04-10 11:32

本文选题：危重症胰腺炎 + p-gsk-3β　；参考：《河北联合大学》2014年硕士论文

【摘要】：目的急危重症胰腺炎（SAP）并发心脏损害的死亡率极高，严重威胁患者的生命。通过分子生物学手段探讨其发生的具体机制，并为开发合理的临床用药提供理论依据。方法本研究利用5%的牛磺胆酸钠建立大鼠SAP模型，将80只健康雄性SD大鼠随机分为两组，假手术组（sham组）及模型组，每组各40只，每组又下设0h、3h、6h、12h及24h不同的时间点，，采用全自动生化仪检测血清胰淀粉酶（AMY）、乳酸脱氢酶（LDH）及肌酸激酶同工酶（CK-MB）的变化，HE染色观察大鼠胰腺及心脏组织的形态学变化，Western blot法及免疫荧光法检测大鼠心肌组织P-GSK-3β的表达情况，线粒体肿胀实验测定大鼠心肌线粒体吸光度的变化，离体大鼠心脏灌流实验观察牛磺胆酸钠对大鼠心肌组织P-GSK-3β的表达影响。结果1.血清酶检测结果显示，与0h组相比，各模型组的胰淀粉酶、乳酸脱氢酶及肌酸激酶同工酶都有所升高，其中以12h升高的最为显著（P 0.05）。2.HE染色结果显示，胰腺：与0h组相比，各模型组胰腺组织都不同程度的损害，3h:间质血管扩张出血，伴有大量炎性细胞浸润，实质变化不明显。6h：大面积的胰腺小叶出血坏死，间质血管扩张出血，并伴有胰管、胆管的出血坏死；12h：出现了边缘小叶及胰管的出血坏死，伴有大量的纤维素渗出；24h：除有胰腺出血坏死的表现外，部分区域的胰腺小叶纤维组织增生明显，小胰管增生显著。心脏：与0h组相比，各模型组心脏组织都出现了不同程度的损害。3h：肌间隙增宽，间质有轻度的血管扩张出血；6h：心肌组织肌间隙增大增宽，间质有少量的毛细血管扩张充血；12h：除肌间隙增宽外，出现了严重的肌肉的排列紊乱现象，部分区域甚至出现了心肌细胞的变性、肿胀，心肌间质血管扩张充血，比3h、6h加重；24h：心肌间隙增宽，出现点状、灶状及片状的炎性细胞浸润，以中性分叶核白细胞及中性单核细胞为主，部分区域的肌间隙里面还有弥漫性分布的炎性细胞，类似心肌炎的表现。3.Western blot及免疫荧光结果显示，与0h组相比，各模型组心肌组织P-GSK-3β的表达都有所增加，且以12h升高的最为明显（P 0.05）。4.线粒体肿胀实验结果显示，各组心肌线粒体吸光度都有不同程度的减少，且以12h减少最多（P 0.05），5.离体大鼠心脏灌流实验结果显示，牛磺胆酸钠灌流离体大鼠心脏后，心肌组织中P-GSK-3β的表达情况与对照组相比，无明显变化。结论1）大鼠急危重症胰腺炎在12h并发的心肌损害最为严重。 2）P-GSK-3参与到急危重症胰腺炎并发的心肌损害作用中。 3）在大鼠急危重症胰腺炎并发的心肌损害中，P-GSK-3水平增加并不抑制mPTP的开放。
[Abstract]:Objective the death rate of severe acute pancreatitis (SAP) complicated with cardiac damage is very high, which is a serious threat to the life of the patients.The mechanism of its occurrence was discussed by molecular biological methods, and the theoretical basis was provided for the development of rational clinical drug use.The changes of serum amylase, lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MBB) were detected by automatic biochemical instrument. The morphological changes of rat pancreas and heart tissue were observed by HE staining. Western blot method and immunofluorescence assay were used to detect myocardial changes in rats.The expression of P-GSK-3 尾 in tissues,Mitochondrial swelling test was used to measure the changes of myocardial mitochondrial absorbance in rats, and the effects of sodium taurocholate on the expression of P-GSK-3 尾 in isolated rat hearts were observed.Result 1.The results of serum enzyme detection showed that the levels of amylase, lactate dehydrogenase and creatine kinase isozymes in each model group were higher than those in 0h group. The results of P 0.05).2.HE staining at 12 h showed that the pancreas: compared with the 0 h group, the serum amylase, lactate dehydrogenase and creatine kinase isozymes in each model group were higher than those in the 0 h group.In each model group, pancreatic tissue was damaged for 3 h in varying degrees: interstitial vasodilation, massive inflammatory cell infiltration, no significant changes in parenchyma, large area of pancreatic lobular hemorrhage, interstitial vascular dilatation and hemorrhage, and pancreatic duct.Heart: compared with 0 h group, each model group showed different degrees of damage. 3 h: muscle space widened, interstitial vasodilation hemorrhage 6 h, myocardial tissue muscle space enlarged and widened.There was a small amount of capillary dilatation in the interstitial for 12 h: in addition to the widening of the muscle space, there was a serious disorder of muscle arrangement, and some areas even showed degeneration, swelling, hyperemia of the myocardial interstitial vessels.Compared with 3 h or 6 h, the myocardial space was widened, and the infiltration of focal and flake inflammatory cells was observed. The majority of them were neutrophilic lobular leukocytes and neutrophil mononuclear cells, and there were diffuse inflammatory cells in some areas of muscle space.The results of mitochondria swelling test showed that the absorbance of myocardial mitochondria in each group was decreased in varying degrees, and the maximum decrease was at 12 hours (P 0.05).The results of isolated rat heart perfusion experiment showed that the expression of P-GSK-3 尾 in myocardium of isolated rat heart after perfusing with sodium taurocholate had no significant change compared with that of control group.Conclusion 1) the myocardial damage of acute severe pancreatitis in rats was the most serious at 12 hours.2)P-GSK-3 is involved in myocardial damage caused by acute and severe pancreatitis.3) the increased level of P-GSK-3 did not inhibit the opening of mPTP.
【学位授予单位】：河北联合大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R576

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