两种大鼠下腔静脉移植模型建立方法的比较分析_任章勇.pdf

2022 年 12 月第 30 卷 第 8 期中国实验动物学报ACTA LABORATORIUM ANIMALIS SCIENTIA SINICADecember 2022Vol.30 No.8任章勇,吕少诚,王芳菲,等.两种大鼠下腔静脉移植模型建立方法的比较分析 J.中国实验动物学报,2022,30(8):1071-1079.Ren ZY,Lyu SC,Wang FF,et al.Comparative analysis of two methods for establishing inferior vena cava transplantation model in rats J.Acta Lab Anim Sci Sin,2022,30(8):1071-1079.Doi:10.3969/j.issn.1005-4847.2022.08.008作者简介任章勇(1994),男,在读博士研究生,研究方向:胰腺恶性肿瘤。Email:13120206720 通信作者贺强(1954),男,主任医师,教授,博士,研究方向:肝脏移植及肝胆胰恶性肿瘤的治疗。Email:heqiang349 两种大鼠下腔静脉移植模型建立方法的比较分析任章勇,吕少诚,王芳菲,赵昕,郎韧,贺强(首都医科大学附属北京朝阳医院肝胆外科,北京 100020)【摘要】目的 本研究拟通过“套管法”和“吻合法”分别建立大鼠静脉置换模型,评估两种模型建立方法的优缺点,为后续静脉置换相关的病理机制研究提供安全有效的静脉置换模型。方法以无特定病原体(specific pathogen free,SPF)级 BN 大鼠作为供体和受体,分别采用套管法和吻合法构建下腔静脉置换模型,分析两组模型术中所用总手术时间和血流阻断时间,术后 48 h 模型存活率,术后每隔 1 周行腹部血管超声观察置换段血管通畅情况及血流速度变化,于术后 1 周、2 周、4 周处死大鼠获取置换段静脉,观察其管径变化,通过组织病理评估血管移植后病理改变特点、qPCR 检测移植段血管组织中 TGF-1的表达水平,评估两种模型建立方法的优缺点。结果 套管法和吻合法各建立大鼠下腔静脉置换模型 18 只,术后 48 h 内成功率无明显差异100.0%(18/18)vs 94.4%(17/18),P=0.310;套管法的总手术时间和血流阻断时间均较吻合法短(35.8 3.6)min vs(56.8 4.9)min,P 0.05;(14.6 2.9)min vs(35.1 4.5)min,P 0.05。腹部血管超声结果显示,移植段静脉血流速度早期血流速度较前有所增大,随着术后时间的延长,血流速度逐渐减小,4 周时套管组明显低于吻合组(P 0.05);术后 4 周时,套管组模型移植段静脉周围可见静脉侧枝循环形成,但移植静脉尚通畅,吻合组 4 周内未见明显静脉侧枝循环形成。随着术后时间的延长,两组模型中移植静脉管壁逐渐增厚直径均逐渐缩小,套管组变化较吻合组更为明显,术后各个时间内其直径均小于吻合组(P 0.05);套管组术后静脉壁外层可见纤维素沉积形成纤维包壳,肌层和内膜增生,管腔狭窄;吻合组模型术后移植静脉壁中单个核细胞及 CD4+T 淋巴细胞浸润较套管组明显(P 0.05);随着术后时间的延长,两组模型移植段静脉壁中 TGF-1的表达逐渐增多,各个时间点内,套管组均高于吻合组(P 0.05)。结论 套管法和吻合法均能安全有效地建立大鼠下腔静脉置换模型。与套管法相比,吻合法建立的静脉置换模型具有更轻的非特异性验证和管壁重塑,远期通畅率更高,更适用于静脉置换相关实验研究。【关键词】静脉置换;静脉移植;内膜增生;动物模型【中图分类号】Q95-33 【文献标识码】A 【文章编号】1005-4847(2022)08-1071-09Comparative analysis of two methods for establishing inferior vena cava transplantation model in ratsREN Zhangyong,LYU Shaocheng,WANG Fangfei,ZHAO Xin,LANG Ren,HE Qiang(Department of Hepatobiliary Surgery,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China)Corresponding author:HE Qiang.E-mail:heqiang349 【Abstract】ObjectiveThis study intended to establish rat venous replacement models through the“cannula method”and“anastomosismethod”and to identify a safe and effective venous replacement model for studies into the pathological mechanisms related to venous replacement.MethodsSPF BN rats were used.The venous replacement models were constructed using the cannula or anastomosis method.The total operation time,blood flow blockage time and the 48 h survival rates after surgery were measured and analyzed.Abdominal vascular ultrasound was performed to observe the patency and blood flow velocity changes.The rats were killed to obtain the graft at different timepoint.Pathological changes were evaluated by histopathology,and TGF-1 expression was detected by q-PCR.Results36 rats with venous 中国实验动物学报 2022 年 12 月第 30 卷第 8 期 Acta Lab Anim Sci Sin,December 2022,Vol.30,No.8replacement were established via the cannula method or anastomosis method.There was no significant difference in the 48-hour survival rates 100.0%(18/18)vs 94.4%(17/18),P=0.310.The total operation time and blood flow blocking time of the cannula method were shorter than that of the anastomosis method (35.8 3.6)min vs(56.8 4.9)min,P 0.05;(14.6 2.9)min vs(35.1 4.5)min,P 0.05.The vein blood flow velocity in the transplantation section increased in the early stage and gradually decreased with postoperative time.At 4 weeks,blood flow velocity in the cannula group was significantly lower than that in the anastomosis group(P 0.05).At 4 weeks after the operation,the formation of venous collateral circulation was seen around the vein of the transplanted section of models in the cannula group,but the transplanted vein was still unobstructed.There was no obvious formation of venous collateral circulation in the anastomotic group within 4 weeks.With the advancement of postoperative time,the wall of the transplanted vein thickened and the diameter reduced gradually in the two groups.The vein diameter was smaller in the cannula group than in the anastomosis group at each timepoint after the operation(P 0.05).In the cannula group,cellulose deposition was observed on the outer layer of the vein wall,forming fiber cladding in the muscle layer and intima hyperplasia and lumen stenosis.The degree of infiltration of mononuclear cells and CD4+T lymphocytes in the transplanted vein wall in the anastomotic group was significantly higher than that in the cannula group(P 0.05).With the advancement of postoperative time,the expression of TGF-1 in the vein wall of the transplanted section of the two groups increased gradually.At each timepoint,TGF-1 expression in the cannula group was higher than that in the anastomosis group(P 0.05).Conclusions Both the cannula and anastomosis method can be used to safely and effectively establish a rat inferior vena cava replacement model.Compared with the cannula method,the vein replacement model established by the anastomosis method has lighter non-specific verification and wall remodeling,and has a higher long-term patency rate,which is more suitable for experime