miR-124在运动干预下负调控Hedgehog通路对OJ致肝损伤的改善作用

miR-124 Improving OJ-induced Liver Injury through Downregulation of Hedgehog Pathway under Exercise Intervention

为探讨运动干预下通过miR-124/Hedgehog传导途径对梗阻性黄疸小鼠致肝损伤的改善作用,该文采用雄性KM小鼠构建胆汁淤积性黄疸模型,随机分成模型组(M)和有氧运动组(TE),设空白组(SO),每组15只。采用一般情况观察、ELISA技术、HE染色、肝组织纤维化及炎性指标检测、qRT-PCR验证研究规律有氧运动的改善作用机制。结果显示,M组多部位黄染,尿液颜色加深,粪便呈浅灰色,解剖观察到其胆管悬挂处呈囊性扩张。且血清TBIL、TBA、ALT、AST、ALP蛋白含量及炎性因子TNF-α、IL-1β、NF-κB p65、MMP-9的含量相比SO组显著升高,抗炎性因子IL-10显著降低(P<0.01)。与M组比较,TE组肝功能指标及炎性因子表达有明显改善,IL-10蛋白含量显著增高,肝湿重下降,肝纤维化指标HA、LN、V-C、PIIINP蛋白含量下降(P<0.01)。镜下观察M组肝索排列紊乱,且空泡、核裂症、变性大量出现,肝纤维化严重,TE组肝细胞损伤症状显著减轻。与M组相比,TE组中miR-124表达显著增高(P<0.01),shh、Ptch-1、Gli1 mRNA表达降低(P<0.01)。提示规律有氧运动可通过上调miR-124负性调控Hedgehog通路中关键基因的表达水平,进而减轻胆汁淤积性黄疸后炎症反应,改善肝损伤。

In order to study the protective effect of aerobic exercise on OJ (obstructive jaundice)-induced liver injury in mice,and explore the possible liver protection mechanism of aerobic exercise from the miR-124-mediated regulation of Hedgehog pathway,KM mice were used to construct a cholestatic jaundice model.They were randomly divided into model group (M),aerobic exercise group (TE),and blank group (SO),with 15 mice in each group.General observation,ELISA technique,HE staining,liver tissue fibrosis and inflammatory indicator detection,qRT-PCR were employed to verify the improvement mechanism of regular aerobic exercise.The results indicated that in M group,many parts were yellow stained,and the urine color became darker.The stool was light gray.After anatomy,cystic dilatation could be observed at the bile duct suspension.Compared with SO group,the contents of TBIL,TBA,ALT,AST,ALP proteins and inflammatory factors TNF-α,IL-1β,NF-κB p65,and MMP-9 in serum were significantly increased,while anti-inflammatory factor IL-10 was significantly reduced (P<0.01).Compared with M group,liver function indicators and inflammatory factor expression in TE group were significantly improved;IL-10 protein content was significantly increased;wet liver weight decreased,and liver fibrosis indicators HA,LN,IV-C,PIIINP protein content decreased (P<0.01).Under the microscope,it could be seen that the liver cords in M group were disorderly arranged;vacuoles,mitosis and degeneration appeared in large number,accompanied with severe liver fibrosis,and the symptoms of liver cell injury in TE group were significantly alleviated.Compared with M group,miR-124 expression in TE group was significantly increased (P<0.01),while shh,Ptch-1,Gli1 mRNA expression was reduced (P<0.01).In conclusion,regular aerobic exercise can alleviate the cholestatic jaundice-induced inflammatory reaction by upregulating miR-124 downregulation of related protein and gene levels of Hedgehog pathway,so as to improve the liver injury.