胆碱能抗炎通路在大鼠胆肠吻合术后吻合口瘢痕形成中的作用

Effect of cholinergic anti-inflammatory pathway in the scar formation after bflioenteric anastomosis inrats

目的研究胆碱能抗炎通路在大鼠胆肠吻合术后局部吻合口瘢痕形成中的作用。方法SD大鼠60只，随机分为10组，每组6只，包括正常组（一期术前组、二期术前组）、实验组（3天组、1周组、2周组、1月组），对照组（3天组、1周组、2周组、1月组）。实验组大鼠术后第1天即开始给予胆碱能抗炎通路药物（山莨菪碱25mg／kg联合新斯的明50μk）腹腔注射，连续7d；对照组给予等量生理盐水。比较两组大鼠不同时间点胆肠吻合口的直径、抗平滑肌抗体（d—SMA）和TGF．131免疫组化染色以及TGF-β1mRNA相对表达量。结果顺利完成各组实验手术操作，建立大鼠模型。（1）对照组大鼠的胆总管直径随着时间的推移而逐渐出现狭窄，术后1个月时为（3．3±0．2）mm，而实验组大鼠的胆总管直径无狭窄趋势，术后1个月时为（4．6±0．2）mm（P〈0．05）。（2）大鼠在胆肠吻合术后，随着时间的推移，吻合口组织中α-SMA表达逐渐增多，而TGF-β1逐渐减少。其中实验组大鼠吻合口的瘢痕组织形成情况在各个时间点均比对照组轻。（3）对照组大鼠术后1个月吻合口组织的TGF-β1相对表达量为（1．02±0．28），实验组大鼠术后1个月为（0．51±0．30，P〈0．05）。结论胆碱能抗炎通路能有效减少大鼠胆肠吻合术后吻合口瘢痕形成，进而减少术后吻合口狭窄并发症的发生。

Objective To evaluate the roles of cholinergic anti-inflammatory pathway in scar forma- tion after bilioenteric anastomosis in rats. Methods A total of 60 SD rats were randomly divided into l0 groups with 6 rats in each group, including normal groups （phase I group and phase Hgroup）, experimental groups （3 days group, 1 week group, 2 weeks group and 1 month group） and control groups （3 days group, 1 week group, 2 weeks group and 1 month group）. The experimental groups have started intraperitoneal injection with agents （ anisodamine 25 mg/kg plus neostigmine 50 ixg/kg ） targeting cholinergic anti-inflammatory path- way for consecutive 7 days since dl post operation. The control groups were given normal saline in equivalent dosage. The diameter of bile duct, ct-SMA and TGF-β1 status tested by immunohistochemical staining, and TGF-β1 relative expression were compared between the two groups at different endpoints. Results The rat models were well estabalished after experimental procedures. （ 1 ） The diameter of common bile duct in the control group was progressively narrowed, and the diameter in 1 month after operation was （3.3 ± 0.2） mm. In contrast, the diameter of common bile duct in the experimental group was not narrowed, which was （ 4.6 ± 0.2） mm in 1 month after operation （ P 〈 O. 05 ）, （2） α-SMA and TGF-β1 were overexpressed in the tissue of bilioenteric anastomosis, and the α-SMA level was gradually elevated, and TGF-β1 expression was gradually decreased. It was observed that collagen fibers proliferated, and the granulation tissue was formed in the bilioenteric anastomosis. The pathological tests of the experimental groups showed more favorable scar formation in the bilioenteric anastomotic tissue than that in the control groups at all endpoints. （3） The relative expression of TGF-β1 gene in the control group in 1 month after operation was 1.02 ±0.28, which was 0.51±0.30 in the experimental group （P 〈 O. 05 ）. Conclusion Cholinergie anti-inflammatory pathway could effectively decrease the complications caused by postoperative anastomosis stenosis through reducing bilioenteric anastomotic scar formation.