大鼠左肺原位肺移植改良模型的学习曲线及步骤解析

唐宏涛; 李偲涵; 王俊杰; 黄桁; 徐琳; 陈婷婷; 刘眉涵; 付思怡; 田东

doi:10.3969/j.issn.1674-7445.2021.05.009

大鼠左肺原位肺移植改良模型的学习曲线及步骤解析

doi: 10.3969/j.issn.1674-7445.2021.05.009

637000 四川南充，川北医学院附属医院心肺移植研究室（唐宏涛、李偲涵、王俊杰、黄桁、徐琳、陈婷婷、刘眉涵、付思怡、田东）；川北医学院附属医院院士（专家）工作站（田东）

基金项目:

医学影像四川省重点实验室开放课题 MIKLSP202007

南充市市校合作科研专项 19SXHZ0189

川北医学院附属医院科研发展计划项目 2021ZK003

2021年四川省大学生创新创业训练计划项目 202110634025

详细信息

通讯作者:
田东，男，博士，副主任医师，研究方向为肺移植基础与临床研究，E-mail: TianD_EATTS@nsmc.edu.cn

中图分类号: R617, R563
计量
- 文章访问数: 814
- HTML全文浏览量: 298
- PDF下载量: 99
- 被引次数: 0
出版历程
- 收稿日期: 2021-05-15
- 网络出版日期: 2021-09-15
- 刊出日期: 2021-09-15

Learning curve and key procedures analysis of modified rat models of orthotopic left lung transplantation

Heart and Lung Transplant Research Laborary, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China

More Information

Corresponding author: Tian Dong, E-mail: TianD_EATTS@nsmc.edu.cn

摘要

摘要: 目的总结和解析大鼠左肺原位肺移植改良模型的关键步骤，为大鼠肺移植模型的建立提供更多经验。方法两名术者(术者A、B)分别连续做完整移植手术15例，并将每5例手术分为1个练习阶段。详细记录各移植程序的操作时间，计算总体成功率和1周存活率在不同练习阶段之间的差异，并采用累积和法构建学习曲线。结果术者A、B首次移植成功的手术次数分别为第5次、第6次，移植总体成功率分别为80%和87%，大鼠1周存活率均为92%。随着手术例数的增加，两名术者的心肺整体获取、套管制备、冷缺血、热缺血、移植和总手术时间均呈下降趋势(均为P < 0.05)。术者A、B分别有3只、2只大鼠因麻醉过度导致心力衰竭死亡，各有1只大鼠分别于术后1 d、7 d死亡，死因分别为静脉扭曲和肺不张。累积和法拟合优度R2分别为0.992 8和0.976 6，两名术者分别于第7例和第8例移植手术时达到学习曲线拐点。结论本课题组改良的大鼠左肺原位肺移植模型在理论和技术上均具有良好的可操作性和可重复性，且具有学习曲线短、手术时间短、受体存活率高且并发症少的优点，在肺移植基础研究领域值得进一步推广和应用。
- 肺移植 /
- 动物模型 /
- 改良模型 /
- 套管技术 /
- 累积和法 /
- 学习曲线 /
- “钟摆式”吻合法 /
- 步骤解析
Abstract: Objective To summarize and analyze the key procedures of the modified rat model of orthotopic left lung transplantation, aiming to provide more experience for the establishment of rat models of lung transplantation. Methods Two surgeons (A and B) performed complete transplantation in consecutive 15 rat models, and every 5 surgeries were divided into 1 practice stage. The operating time of each transplantation procedure was recorded. The differences of overall success rate and 1-week survival rate were calculated among different practice stages. The learning curve was delineated by the cumulative sum method. Results For surgeons A and B, the number of the first successful transplantation was the 5th and 6th time, the overall success rates of transplantation were 80% and 87% respectively, and the 1-week survival rates of rats both were 92%. Along with the increasing number of surgeries, the entire cardiopulmonary procurement, cannula preparation, cold ischemia, warm ischemia, transplantation and total operation time by two surgeons showed a significantly downward trend (all P < 0.05). For surgeons A and B, 3 and 2 rats died of heart failure due to overdose anesthesia, and 1 rat died of vein distortion at postoperative 1 d and 1 died of atelectasis at postoperative 7 d, respectively. The goodness of fit (R²) of cumulative sum method was 0.992 8 and 0.976 6. The turning point of learning curve was achieved in the 7th and 8th transplantation for surgeons A and B. Conclusions The modified rat model of orthotopic left lung transplantation yields high operability and repeatability both theoretically and technologically, and has multiple advantages of short learning curve, short operation time, high survival rate of the recipients and few complications, which is worthy of application in the basic research of lung transplantation.
- Lung transplantation /
- Animal model /
- Modified model /
- Cannula technique /
- Cumulative sum method /
- Learning curve /
- "Pendulum" anastomosis /
- Procedure analysis

HTML全文

下载: 全尺寸图片幻灯片

图 1 大鼠左肺原位肺移植手术步骤

注：A图示气管插管；B图示供肺灌注；C~E图示游离左肺及制备套管；F~I图示受体移植。

Figure 1. Surgical procedures for orthotopic left lung transplantation in rats

下载: 全尺寸图片幻灯片

图 2 术者A、B的CUSUM学习曲线

注：A图为术者A的CUSUM学习曲线，B图为术者B的CUSUM学习曲线。

Figure 2. The CUSUM learning curves of surgeon A and B

下载: 全尺寸图片幻灯片

表 1 两名术者肺移植各阶段操作时间

Table 1. The operating time of each stage of lung transplantation from two surgeons (x±s, min)

移植程序时间	术者A				术者B
移植程序时间	阶段Ⅰ	阶段Ⅱ	阶段Ⅲ	P值	阶段Ⅰ	阶段Ⅱ	阶段Ⅲ	P值
心肺整体获取时间	11.4±1.8	10.6±1.4	8.8±0.7 ^a	0.006	16.9±1.6	16.0±0.8	12.2±1.7 ^{a, b}	< 0.001
套管制备时间	23.5±5.9	25.1±2.8	17.8±3.1 ^{a, b}	0.038	48.9±2.1	39.2±7.3 ^a	22.4±6.2 ^{a, b}	< 0.001
冷缺血时间	52.9±8.8	51.7±2.8	43.7±2.6 a, ^b	0.017	84.9±1.2	69.9±12.7 ^a	43.8±3.4 a, ^b	< 0.001
热缺血时间	41.2±11.3	23.6±7.5 ^a	13.1±3.5 ^a	< 0.001	34.1±2.0	33.0±7.0	21.3±11.1 ^{a, b}	0.039
移植时间	73.2±8.5	53.0±6.2 ^a	42.2±3.0 ^{a, b}	< 0.001	74.0±2.1	66.9±11.4	44.2±11.3 ^{a, b}	0.001
总手术时间	108.8±10.8	88.8±9.0 ^a	68.7±4.5 ^{a, b}	< 0.001	139.8±4.3	122.1±14.1 ^a	80.7±14.2 ^{a, b}	< 0.001
注：与阶段Ⅰ比较，^aP < 0.05/3；与阶段Ⅱ比较，^bP < 0.05/3。

下载: 导出CSV

参考文献(28)

[1]	KULKARNI HS, CHERIKH WS, CHAMBERS DC, et al. Bronchiolitis obliterans syndrome-free survival after lung transplantation: an International Society for Heart and Lung Transplantation Thoracic Transplant Registry analysis[J]. J Heart Lung Transplant, 2019, 38(1): 5-16. DOI: 10.1016/j.healun.2018.09.016.
[2]	SATO M, TIAN D. Preface: strategies to achieve long-term success of lung transplantation[J]. Ann Transl Med, 2020, 8(6): 406. DOI: 10.21037/atm.2020.02.160.
[3]	TIAN D, HUANG H, WEN HY. Noninvasive methods for detection of chronic lung allograft dysfunction in lung transplantation[J]. Transplant Rev (Orlando), 2020, 34(3): 100547. DOI: 10.1016/j.trre.2020.100547.
[4]	TIAN D, WANG Y, SHⅡYA H, et al. Outcomes of marginal donors for lung transplantation after ex vivo lung perfusion: a systematic review and Meta-analysis[J]. J Thorac Cardiovasc Surg, 2020, 159(2): 720-730. DOI: 10.1016/j.jtcvs.2019.07.087.
[5]	严浩吉, 陈蔚洋, 魏桢婷, 等. 肺移植术后胃肠道并发症的研究进展[J]. 器官移植, 2020, 11(6): 749-753, 764. DOI: 10.3969/j.issn.1674-7445.2020.06.016. YAN HJ, CHEN WY, WEI ZT, et al. Research progress on gastrointestinal complications after lung transplantation[J]. Organ Transplant, 2020, 11(6): 749-753, 764. DOI: 10.3969/j.issn.1674-7445.2020.06.016.
[6]	RUIZ-PÉREZ D, LARGO C, GARCÍA-RÍO F. Technical aspects and benefits of experimental mouse lung transplantation[J]. Arch Bronconeumol, 2016, 52(12): 596-604. DOI: 10.1016/j.arbres.2016.04.008.
[7]	MIZUTA T, KAWAGUCHI A, NAKAHARA K, et al. Simplified rat lung transplantation using a cuff technique[J]. J Thorac Cardiovasc Surg, 1989, 97(4): 578-581.
[8]	WANG J, TAN J, LIU Y, et al. Amelioration of lung ischemia-reperfusion injury by JNK and p38 small interfering RNAs in rat pulmonary microvascular endothelial cells in an ischemia-reperfusion injury lung transplantation model[J]. Mol Med Rep, 2018, 17(1): 1228-1234. DOI: 10.3892/mmr.2017.7985.
[9]	TANE S, NODA K, TOYODA Y, et al. Bronchial-arterial-circulation-sparing lung preservation: a new organ protection approach for lung transplantation[J]. Transplantation, 2020, 104(3): 490-499. DOI: 10.1097/TP.0000000000002984.
[10]	GAO W, MENG QM, CUI XG. Budesonide instillation immediately after reperfusion ameliorates ischemia/reperfusion-induced injury in the transplanted lung of rat[J]. Exp Lung Res, 2017, 43(9/10): 439-446. DOI: 10.1080/01902148.2017.1405103.
[11]	SCHOSSLEITNER K, HABERTHEUER A, FINSTERWALDER R, et al. A peptide to reduce pulmonary edema in a rat model of lung transplantation[J]. PLoS One, 2015, 10(11): e0142115. DOI: 10.1371/journal.pone.0142115.
[12]	LAMA VN, BELPERIO JA, CHRISTIE JD, et al. Models of lung transplant research: a consensus statement from the National Heart, Lung, and Blood Institute workshop[J]. JCI Insight, 2017, 2(9): e93121. DOI: 10.1172/jci.insight.93121.
[13]	OHSUMI A, KANOU T, ALI A, et al. A method for translational rat ex vivo lung perfusion experimentation[J]. Am J Physiol Lung Cell Mol Physiol, 2020, 319(1): L61-L70. DOI: 10.1152/ajplung.00256.2019.
[14]	BURKI S, NODA K, PHILIPS BJ, et al. Impact of triptolide during ex vivo lung perfusion on grafts after transplantation in a rat model[J]. J Thorac Cardiovasc Surg, 2020, DOI: 10.1016/j.jtcvs.2019.12.104[Epubahead of print].
[15]	RAJAB TK. Anastomotic techniques for rat lung transplantation[J]. World J Transplant, 2018, 8(2): 38-43. DOI: 10.5500/wjt.v8.i2.38.
[16]	HABERTHEUER A, KOCHER A, LAUFER G, et al. Innovative, simplified orthotopic lung transplantation in rats[J]. J Surg Res, 2013, 185(1): 419-425. DOI: 10.1016/j.jss.2013.05.006.
[17]	TIAN D, SHⅡYA H, SATO M, et al. Rat lung transplantation model: modifications of the cuff technique[J]. Ann Transl Med, 2020, 8(6): 407. DOI: 10.21037/atm.2020.02.46.
[18]	GAUTHIER JM, RUIZ-PÉREZ D, LI W, et al. Diagnosis, pathophysiology and experimental models of chronic lung allograft rejection[J]. Transplantation, 2018, 102(9): 1459-1466. DOI: 10.1097/TP.0000000000002250.
[19]	HAAM S, NODA K, PHILIPS BJ, et al. Cyclosporin A administration during ex vivo lung perfusion preserves lung grafts in rat transplant model[J]. Transplantation, 2020, 104(9): e252-e259. DOI: 10.1097/TP.0000000000003237.
[20]	KANOU T, OHSUMI A, KIM H, et al. Inhibition of regulated necrosis attenuates receptor-interacting protein kinase 1-mediated ischemia-reperfusion injury after lung transplantation[J]. J Heart Lung Transplant, 2018, 37(10): 1261-1270. DOI: 10.1016/j.healun.2018.04.005.
[21]	DIANA P, ZAMPIERI D, FURLANI E, et al. Lung ultrasound as a monitoring tool in lung transplantation in rodents: a feasibility study[J]. J Thorac Dis, 2018, 10(7): 4274-4282. DOI: 10.21037/jtd.2018.06.52.
[22]	GUO H, NIE J, FAN K, et al. Improvements of surgical techniques in a rat model of an orthotopic single lung transplant[J]. Eur J Med Res, 2013, 18(1): 1. DOI: 10. 1186/2047-783X-18-1.
[23]	BALZER C, CLEVELAND WJ, JINKA TR, et al. Video laryngoscopic oral intubation in rats: a simple and effective method[J]. Am J Physiol Lung Cell Mol Physiol, 2020, 318(5): L1032-L1035. DOI: 10.1152/ajplung.00498.2019.
[24]	KONNO K, SHIOTANI Y, ITANO N, et al. Visible, safe and certain endotracheal intubation using endoscope system and inhalation anesthesia for rats[J]. J Vet Med Sci, 2014, 76(10): 1375-1381. DOI: 10.1292/jvms.14-0146.
[25]	PIERÓG J, TAMO L, FAKIN R, et al. Bone marrow stem cells modified with human interleukin 10 attenuate acute rejection in rat lung allotransplantation[J]. Eur J Cardiothorac Surg, 2018, 53(1): 194-200. DOI: 10.1093/ejcts/ezx257.
[26]	SAITO M, CHEN-YOSHIKAWA TF, SUETSUGU K, et al. Pirfenidone alleviates lung ischemia-reperfusion injury in a rat model[J]. J Thorac Cardiovasc Surg, 2019, 158(1): 289-296. DOI: 10.1016/j.jtcvs.2018.08.098.
[27]	RAJAB TK. Techniques for lung transplantation in the rat[J]. Exp Lung Res, 2019, 45(9/10): 267-274. DOI: 10.1080/01902148.2019.1675806.
[28]	GÓMEZ-HERNÁNDEZ MT, NOVOA NM, VARELA G, et al. Quality control in anatomical lung resection. major postoperative complications vs failure to rescue[J]. Arch Bronconeumol (Engl Ed), 2021, 57(4): 251-255. DOI: 10.1016/j.arbres.2019.12.009.