中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (14): 2152-2157.doi: 10.12307/2023.053

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

建立适用于BALB/c Nude小鼠的急性肝衰竭模型

谈佳音，刘  阳，李  斌，陆  利，宋慧芳   

  1. 山西医科大学人体解剖学教研室，山西省太原市  030001
  • 收稿日期:2022-01-29 接受日期:2022-03-10 出版日期:2023-05-18 发布日期:2022-09-30
  • 通讯作者: 宋慧芳，博士，副教授，山西医科大学人体解剖学教研室，山西省太原市 030001 陆利，博士，教授，山西医科大学人体解剖学教研室，山西省太原市 030001
  • 作者简介:谈佳音，女，1997年生，河北省石家庄市人，汉族，山西医科大学在读硕士，主要从事干细胞衰老及心脏、肝脏修复方向研究工作。
  • 基金资助:
    山西省自然科学研究面上项目(20210302123296)，项目负责人：宋慧芳；山西省回国留学人员科研资助项目(2021-078)，项目负责人：宋慧芳；中国博士后科学基金面上资助项目(2021M691994)，项目负责人：宋慧芳

Establishment of an acute liver failure model in BALB/c Nude mice

Tan Jiayin, Liu Yang, Li Bin, Lu Li, Song Huifang   

  1. Department of Human Anatomy, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Received:2022-01-29 Accepted:2022-03-10 Online:2023-05-18 Published:2022-09-30
  • Contact: Song Huifang, MD, Associate professor, Department of Human Anatomy, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China Lu Li, MD, Professor, Department of Human Anatomy, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • About author:Tan Jiayin, Master candidate, Department of Human Anatomy, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Supported by:
    Natural Science Research General Project of Shanxi Province, No. 20210302123296 (to SHF); Shanxi Provincial Research Funding Project for Returned Overseas Students No. 2021-078 (to SHF); General Project of China Postdoctoral Science Foundation, No. 2021M691994 (to SHF)

PDF

114

摘要：

文题释义：
急性肝衰竭：是多种因素引起的急性严重肝脏损害，导致其合成、解毒、排泄和生物转化等功能发生严重障碍或失代偿，出现以凝血机制障碍和黄疸、肝性脑病、腹水等为主要表现的一组临床症候群。
BALB/c Nude小鼠：是一种无胸腺小鼠，免疫力和耐受低下，在进行干细胞研究中常关注干细胞免疫原性问题，为排除干细胞免疫原性引起受体的免疫反应对功能修复等实验结果的影响，通常会用到免疫缺陷的BALB/c Nude小鼠来进行细胞移植实验。

背景：在干细胞和组织工程领域的动物实验中，免疫缺陷裸小鼠(BALB/c Nude小鼠)对于提升研究准确性具有重要意义。但CCl4致BALB/c Nude小鼠发生急性肝衰竭的浓度及检测时间尚不明确，严重影响该动物模型在具有免疫原性的干细胞治疗研究中的应用。
目的：通过与C57小鼠急性肝衰竭模型比较，探索腹腔注射CCl4建立BALB/c Nude小鼠急性肝衰竭模型的合适浓度和检测时间。
方法：将50%浓度的CCl4按照4 mL/kg腹腔注射到C57小鼠(n=6)和BALB/c Nude小鼠(n=6)体内，注射后72 h内观察小鼠存活情况。分别将50%，20%，10%浓度的CCl4按照4 mL/kg腹腔注射到BALB/c Nude小鼠体内，注射后72 h观察小鼠存活情况，并进行肝脏病理切片形态学观察。根据建立两种小鼠急性肝衰竭模型的最佳CCl4浓度，将50%浓度的CCl4腹腔注射到C57小鼠体内，将20%浓度的CCl4腹腔注射到BALB/c Nude小鼠体内，注射前后检测小鼠血清中谷丙转氨酶和谷草转氨酶水平。
结果与结论：①CCl4注射24 h后，C57小鼠的存活率为50%，BALB/c Nude小鼠全部死亡，提示现有针对C57小鼠的最适 CCl4浓度不适合BALB/c Nude小鼠造模；②注射50%浓度CCl4的BALB/c Nude小鼠在24 h之内陆续死亡；注射体积分数20% CCl4的小鼠24 h后存活率是66.7%，至注射后72 h存活率仍为66.7%；注射10%浓度CCl4的小鼠至注射后72 h全部存活；注射后24 h肝病病理结果显示，CCl4注射浓度越小肝脏损伤程度越轻，其中20%浓度的CCl4最适合建立BALB/c Nude小鼠急性肝衰竭模型；③C57小鼠腹腔注射CCl4 4 h后谷丙转氨酶和谷草转氨酶水平达到最高，1 d时开始下降；BALB/c Nude小鼠在腹腔注射CCl4 1 d后谷丙转氨酶和谷草转氨酶水平达到最高，3 d时开始下降；④结果表明，为提高动物实验可靠性，使用C57小鼠建立急性肝衰竭模型时应选择50%浓度的CCl4，在注射后4 h施行干预措施；使用BALB/c Nude小鼠建立急性肝衰竭模型时应选择20%浓度的CCl4，在注射后1 d施行干预措施。

https://orcid.org/0000-0001-7690-3116(谈佳音)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 四氯化碳, 急性肝衰竭, BALB/c Nude小鼠, C57小鼠, 模型

Abstract: BACKGROUND: In animal experiments regarding stem cells and tissue engineering, immunodeficient nude mice (BALB/c Nude) are of great significance for improving research accuracy. However, the CCl4 concentration and detection time for establishing an acute liver failure model in BALB/c Nude mice are still unclear, which seriously affects the application of this animal model in the study of immunogenic stem cell therapy.
OBJECTIVE: To explore the appropriate CCl4 concentration and detection time for establishing the acute liver failure model in BALB/c Nude mice via intraperitoneal injection in comparison with the C57 model of acute liver failure. 
METHODS: CCl4 at the concentration of 50% was intraperitoneally injected into C57 (n=6) and BALB/c Nude mice (n=6) at the dose of 4 mL/kg. The survival of mice was observed within 72 hours after injection. Then, CCl4 at the concentration of 50%, 20%, and 10% was injected intraperitoneally into BALB/c Nude mice at the dose of 4 mL/kg. The survival of mice was also observed within 72 hours after injection and pathological sections of the liver were made for histopathological observation. According to the optimal concentration of CCl4 in two kinds of mouse models of acute liver failure, 50% CCl4 was injected intraperitoneally into C57 mice and 20% CCl4 was injected intraperitoneally into BALB/c Nude mice. The levels of alanine aminotransferase and aspartate aminotransferase in serum of two kinds of mice were detected before and after injection.
RESULTS AND CONCLUSION: The survival rates of C57 and BALB/c Nude mice were 50% and 0 respectively at 24 hours after CCl4 injection, indicating that the optimal concentration of CCl4 is inconsistent in C57 and BALB/c Nude mice. BALB/c Nude mice injected with 50% CCl4 died in succession within 24 hours. The survival rate of BALB/c Nude mice injected with 20% CCl4 was 66.7% after 24 hours and was still 66.7% after 72 hours. All BALB/c Nude mice injected with 10% CCl4 survived until 72 hours after injection. Pathological results of the BALB/c Nude mouse liver 24 hours after injection showed that the lower concentration of CCl4 indicated the less liver damage, and 20% CCl4 was the most suitable for establishing the acute liver failure model in BALB/c Nude mice. In C57 mice undergoing intraperitoneal injection of CCl4, the levels of alanine aminotransferase and aspartate aminotransferase reached the peak at 4 hours and began to decline at 1 day. In BALB/c Nude mice undergoing intraperitoneal injection of CCl4, the levels of alanine aminotransferase and aspartate aminotransferase reached the peak at 1 day and began to decline at 3 days. All these findings indicate that to improve the reliability of animal experiments, 50% CCl4 should be selected when making the acute liver failure model in C57 mice, and intervention measures should be taken 4 hours after injection. When BALB/c Nude mice are used to make the acute liver failure model, 20% CCl4 should be selected and intervention measures should be taken at 1 day after injection.

Key words: carbon tetrachloride, acute liver failure, BALB/c Nude mouse, C57 mouse, model

中图分类号: