胆汁外引流对重症急性胰腺炎大鼠胰腺的保护作用及机制的实验研究

发布时间：2018-10-15 18:18

【摘要】：目的:研究“胆汁外引流”对由4%牛磺胆酸钠方法构建大鼠重症急性胰腺炎(Severe acute pancreatitis,SAP)模型的保护作用及机制。观察胰腺组织内核转录因子Kappa B(NF-κB)蛋白的表达,血清内血红素氧合酶1(HO-1)、高迁移率族蛋白1(HMGB-1)、肿瘤坏死因子-α(TNF-α)和白介素-10(IL-10)的含量以及胰腺组织病理学变化。方法:健康雌性(Sprague-Dawley,SD)大鼠108只,采用逆行十二指肠穿刺胰胆管途径注射4%牛磺胆酸钠方法构建大鼠SAP模型。实验动物分为假手术组和模型组:(1)假手术组分为假手术未引流组(Sham Operation Group,SOG组)和假手术引流组(Sham Drainage Group,SDG组),每组18只。(2)模型组分为模型未引流组(Severe Acute Pancreatitis,SAP组)、模型引流组(Bile Drainage Group,BDG组)每组18只。(3)各组于建模成功后的第3h、6h、12h将大鼠(各时间点n=6)再次麻醉后开腹观察,并取血和胰腺等组织,检测血清AMY的指标,采用酶联免疫法(ELISA)检测血清细胞因子TNF-α、HO-1、HMGB1和IL-10的表达和含量,采用蛋白免疫印迹法(Western Blot)测定胰腺组织内NF-κB P65蛋白表达。通过HE染色大鼠胰腺组织观察其病理学变化。结果:建模后3h、6h、12h:(1)BDG组血清AMY、TNF-α的水平均低于SAP组,差异有统计学意义(P0.05);(2)BDG组的血清HMGB1水平低于SAP组,在6h时,差异有统计学意义(P0.05);(3)BDG组血清IL-10的水平与SAP组比较,差异无统计学意义(P0.05);(4)BDG组血清HO-1水平高于SAP组,差异有统计学意义(P0.05)。BDG组胰腺组织内NF-κB mRNA蛋白表达水平低于SAP组,在3h、6h时有统计学意义(P0.05)。BDG组的病理评分值低于SAP组,差异有统计学意义(P0.05);并且随着时间的推移,大鼠病理评分值呈增高趋势。结论:(1)逆行胰胆管穿刺建立大鼠SAP模型是一种简单操作、重复性好,具有一定稳定性的建模方法,可以满足胆汁外引流对胰腺功能保护作用研究的动物实验需要。(2)胆汁外引流提高SAP模型大鼠的抗炎细胞因子的表达,并且显著抑制机体促炎细胞因子的释放,缓解了胰腺组织的进展。(3)胆汁外引流能改善SAP模型大鼠胰腺组织炎症状况下的病理变化,减轻胰腺组织损伤。
[Abstract]:Aim: to study the protective effect and mechanism of bile drainage on the establishment of severe acute pancreatitis (Severe acute pancreatitis,SAP) model in rats by 4% sodium taurocholate. The expression of Kappa B (NF- 魏 B protein, the contents of heme oxygenase 1 (HO-1), high mobility group protein 1 (HMGB-1), tumor necrosis factor- 伪 (TNF- 伪) and interleukin-10 (IL-10), and the histopathological changes of pancreas were observed. Methods: the rat model of SAP was established by retrograde injection of 4% sodium taurocholate into the pancreaticobiliary tract of 108 healthy female (Sprague-Dawley,SD) rats. The experimental animals were divided into sham-operation group and model group: (1) sham-operation group (Sham Operation Group,SOG group) and sham-operation drainage group (Sham Drainage Group,SDG group). (2) Model group was divided into model undrained group (Severe Acute Pancreatitis,SAP group), model drainage group (Sham Drainage Group,SDG group). Group (Bile Drainage Group,BDG (n = 18) each group (n = 18). (3) the rats (n = 6) were observed in each group after anaesthesia for 12 h at the 3rd hour after successful modeling. Blood and pancreas tissues were taken to detect serum AMY, the expression and content of serum cytokines TNF- 伪, HO-1,HMGB1 and IL-10 were detected by enzyme linked immunosorbent assay (ELISA), and the expression of NF- 魏 B p65 protein in pancreatic tissue was detected by Western blot (Western Blot). The pathological changes of pancreas were observed by HE staining. Results: (1) the level of serum AMY,TNF- 伪 in BDG group was significantly lower than that in SAP group (P0.05); (2) the level of serum HMGB1 in BDG group was lower than that in SAP group. At 6 h, the difference was statistically significant (P0.05); (3) the level of serum IL-10 in BDG group was higher than that in SAP group. There was no significant difference (P0.05); (4) the level of serum HO-1 in BDG group was higher than that in SAP group, and the difference was statistically significant (P0.05) the expression of NF- 魏 B mRNA protein in pancreatic tissue of). BDG group was lower than that of SAP group, and there was statistical significance at 3 h after 6 h (P05). BDG group was lower than SAP group, P 0.05). The difference was statistically significant (P0.05), and with the passage of time, the pathological score of rats showed an increasing trend. Conclusion: (1) the establishment of rat SAP model by retrograde cholangiopancreatopancreatic puncture is a simple, reproducible and stable method. (2) Bile external drainage can increase the expression of anti-inflammatory cytokines and inhibit the release of pro-inflammatory cytokines in SAP model rats. (3) Bile drainage can improve the pathological changes of pancreatic tissue inflammation and alleviate pancreatic tissue injury in SAP model rats.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R576

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