摘要
目的探讨微生态制剂通过下调小鼠肝脏γδT细胞内IL-17的分泌缓解胆汁淤积性肝病的机制。方法随机选取10只正常环境饲养的小鼠作为对照组,每日上下午给予10 mL/kg的生理盐水灌胃共20 d;按相关文献中小鼠胆汁淤积性肝病模型制备方法,将其余用于造模的30只小鼠每日上下午均采用50%葡萄糖+100%猪油脂1︰1比例的混合液10 mL/kg灌胃20 d,第21天,将α-萘基异硫氰酸盐(ANIT)100 mg/kg一次性灌胃,灌胃前后12 h禁食不禁水;随后在完成造模的30只小鼠中,随机选取10只作为模型组,再选取10只作为给药低剂量组,剩余10只为给药高剂量组;自第21天开始,对照组及模型组小鼠每日下午给予10 mL/kg生理盐水灌胃7 d,给药低剂量组以含双歧杆菌活菌数为103 CFU/mL的稀释液10 mL/kg灌胃7 d,给药高剂量组以含双歧杆菌活菌数为107 CFU/mL的稀释液10 mL/kg灌胃7 d。将小鼠处死后,小鼠肝脏HE染色进行组织病理学观察,小鼠血清采用ELISA试验进行肝功能及炎症因子IL-17检测;分离纯化小鼠γδT细胞并进行培养,通过qRT-PCR试验检测IL-17信号通路相关因子IL-17a、IL-17f、CCR6和CD163的表达水平。结果小鼠肝脏组织病理学观察显示微生态制剂对胆汁淤积性肝病有明显缓解效果,并且高剂量比低剂量的效果更明显;血清肝功能检测结果显示模型组小鼠血清ALT、AST、ALP、TBA、TBIL和LP-X水平较对照组升高(t=15.670、13.470、14.170、7.376、7.839、10.430,均P<0.001),血清LEC、CHOL水平降低(t=4.089、6.022,均P<0.001);给药低剂量组及给药高剂量组小鼠血清ALT、AST、ALP、TBA、TBIL和LP-X水平较模型组降低(F=83.330、39.400、71.680、20.820、17.000、27.300,均P<0.001),血清LEC、CHOL水平升高(F=6.419,P=0.520;F=8.486,P=0.014);小鼠血清IL-17检测结果显示模型组较对照组升高(t=42.010,P<0.001),给药低剂量组及给药高剂量组小鼠血清IL-17水平较模型组降低(F=454.700,P<0.001);qRT-PCR�
Objective To explore the mechanisms of microecological preparations alleviating cholestatic liver disease by down-regulating IL-17 secretion inγδT cells of mouse liver.Methods Mouse models of cholestatic liver disease were constructed and divided in control group,model group,low-dose group or high-dose group.After the mice were sacrificed,histopathological observation was performed on mouse liver,and mouse serum was tested for liver function and inflammatory factor IL-17 by ELISA.TheγδT cells in mouse liver were isolated and cultured;The expression levels of IL-17 a,IL-17 f,CCR6 and CD163 related with IL-17-axis were detected with qRT-PCR.Results The histopathological observation showed that the microecological preparation played a significantly beneficial effects on cholestatic liver disease,and the effect of high dose was better than that of low dose.The results of liver function detection showed the levels of ALT,AST,ALP,TBA,TBIL and LP-X in model group were significantly higher than those in the control group(t=15.670,t=13.470,t=14.170,t=7.376,t=7.839,t=10.430;all P<0.001);the levels of LEC and CHOL significantly reduced(t=4.089,t=6.022,all P<0.001).The levels of ALT,AST,ALP,TBA,TBIL and LP-X in the low-dose group and high-dose group were significantly lower than those in the model group(F=83.330,F=39.400,F=71.680,F=20.820,F=17.000,F=27.300;all P<0.001),and the levels of LEC and CHOL significantly increased(F=6.419,P=0.520;F=8.486,P=0.014).The level of IL-17 in the model group was significantly higher than that in the control group(t=42.010,P<0.001).The levels of IL-17 in the low-dose and high-dose groups were significantly lower than that in the model group(F=454.700,P<0.001).The qRT-PCR results showed the expressions of IL-17 a,IL-17 f,CCR6 and CD163 which were main factors related with IL-17 axis were significantly decreased by microecological preparation.Conclusion Microecological preparations can alleviate cholestatic liver disease by down-regulating IL-17 inγδT cells of mouse liver.
作者
李翠茹
武丽
张艳丽
张沙沙
周红霞
LI Cui-ru;WU Li;ZHANG Yan-li;ZHANG Sha-sha;ZHOU Hong-xia(Department of Severe Liver Disease,Handan Infectious Disease Hospital,Handan,Hebei 056000,China;不详)
出处
《中国微生态学杂志》
CAS
CSCD
2021年第2期138-143,共6页
Chinese Journal of Microecology
基金
邯郸市科学技术研究与发展计划(1823208097ZC)。
