建立适用于BALB/c Nude小鼠的急性肝衰竭模型

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

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CCl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CCl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CCl按照4 m L/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CCl按照4 m L/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CCl浓度,将50%浓度的CCl腹腔注射到C57小鼠体内,将20%浓度的CCl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:(1)CCl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CCl浓度不适合BALB/c Nude小鼠造模;(2)注射50%浓度CCl的BALB/c Nude小鼠在24 h之内陆续死亡;注射体积分数20%CCl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CCl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,CCl注射浓度越小肝脏损伤程度越轻,其中20%浓度的CCl最适合建立BALB/c Nude小鼠急性肝衰竭模型;(3)C57小鼠腹腔注射CCl4h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CCl1 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CCl,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CCl,在注射后1 d施行干预措施。

BACKGROUND:In animal experiments rega rding stem cells and tissue engineering,immunodeficient nude mice(BALB/c N ude) are of great significance for improving research accura cy.However,the CClconcentration and detection time for establishing an acute liver failu re 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 explo re the appropriate CClconcentration and detection time for establishing the acute liver failu re model in BALB/c Nude mice via intraperitoneal injection in comparison with the C57 model of acute liver failu re.METHODS:CClat the concentration of 50% was intrape ritoneally injected into C57(n=6) and BALB/c Nude mice(n=6) at the dose of 4 m L/kg.The survival of mice was observed within 72 hours after injection.Then,CClat the concentration of 50%,20%,and 10% was injected intraperitoneally into BALB/c Nude mice at the dose of 4 m L/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 CClin two kinds of mouse models of acute liver failure,50% CClwas injected intraperitoneally into C57 mice and 20% CClwas 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 CClinjection,indicating that the optimal concentration of CClis inconsistent in C57 and BALB/c Nude mice.BALB/c Nude mice injected with 50% CCldied in succession within 24 hours.The survival rate of BALB/c Nude mice injected with 20% CClwas 66.7% after 24 hours and was still 66.7% after 72 hours.All BALB/c Nude mice injected with 10%CClsurvived until 72 hours after injection.Pathological res ults of the BALB/c Nude mouse liver 24 hours after injection showed that the lower concentration of CClindicated the less liver damage,and 20% CClwas the most suitable for establishing the acute liver failure model in BALB/c Nude mice.In C57 mice undergoing intra peritoneal injection of CCl,the levels of alanine aminotransfe rase and aspartate aminotransferase reached the peak at 4 hours and began to decline at 1 day.In BALB/c N ude mice undergoing intrape ritoneal injection of CCl,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 expe riments,50% CClshould 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% CClshould be selected and inte rvention measures should be taken at 1 day after injection.