滋水涵木利胆方改善原发性胆汁性胆管炎小鼠模型肝内胆管上皮细胞间质化的分子机制研究OA

Objective To study the molecular mechanism of Zishui Hanmu Lidan Decoction(ZHLD)in improving epithelial mesenchymal transition of intrahepatic bile duct epithelial cells(IBEC-EMT)in a mouse model of primary biliary cholangitis(PBC).Methods Sixty C57BL/6 female mice were randomly divided into blank group,PBC model group(PBC group),ursodeoxycholic acid group(UDCA group),low dose group(L-ZHLD group),medium dose group(M-ZHLD group)and high dose group(H-ZHLD group),10 in each group.Except for the blank group,the other five groups established a PBC mouse model.At the same time,the same daily dose of normal saline was given to the PBC group by gastrogavage.UDCA group was given ursodeoxycholic acid capsule at 0.1 g/(kg·d)by gastrogavage.The L-ZHLD,M-ZHLD,and H-ZHLD groups were given ZHLD granule at 1,2,and 4 g/(kg·d)by gastrogavage after dilution.Serum and liver tissues were collected after 8 weeks.The expressions of serum antibodies anti-mitoehondrial M2 antibodies(AMA-M2)and anti-nuclear antibody(ANA)were detected by ELISA.HE staining was used to observe the pathological changes of liver tissue.Immunohistochemical staining was used to detect the expressions of cytokeratin(CK)7,CK19,Vimentin,and E-cadherin in bile duct epithelial cells.Western Blot was used to detect the expressions of p-ERK1/2,p-P38(MAPK),p-JNK,E-cadherin,and Vimentin in bile duct epithelial mesenchymal proteins.RT-PCR was used to detect the mRNA quantification of E-cadherin and Vimentin.Results Compared with the blank group,the PBC group showed proliferation of small bile ducts in the portal area,infiltration of lymphocytes and monocytes,increased expressions of CK7,CK19,p-P38MAPK,p-ERK1/2,and p-JNK proteins,increased protein and gene expressions of Vimentind,and decreased protein and gene expressions of E-Cadherin(P＜0.01).Compared with the PBC group,the infiltration of lymphocytes in the portal area decreased in the UDCA group,L,M,and H-ZHLD groups,and the expressions of CK7,CK19,and p-ERK1/2 proteins decreased.Vimentind protein and gene expressions decreased,while E-Cadherin protein and gene expressions increased(P＜0.05,P＜0.01).The expressions of p-P38MAPK and p-JNK proteins decreased in the M and H-ZHLD groups(P＜0.01).Compared with the UDCA group,the infiltration of lymphocytes in the portal area was reduced in the L-,M-,and H-ZHLD groups,CK7 and p-ERK1/2 protein expression were also reduced(P＜0.01).The expressions of CK19,p-P38MAPK,and p-JNK proteins decreased in the M-and H-ZHLD groups,while the protein and gene expressions of Vimentin decreased.The protein and gene expressions of E-Cadherin increased(P＜0.01).Compared with the L-ZHLD group,CK7,CK19,p-P38MAPK,p-JNK and p-ERK1/2 protein expression were reduced in the M-and H-ZHLD groups(P＜0.01);The protein and gene expressions of Vimentin decreased(P＜0.01).The protein and gene expressions of E-Cadherin increased(P＜0.01).Conclusions ZHLD reduced the expressions of CK7,CK19 and Vimentin,up-regulated the expression of E-Cadherin,and reduced bile duct hyperplasia possibly by down-regulating the phosphorylation levels of P38-MAPK,ERK1/2,and JNK pathways,thereby regulating the IBEC-EMT of the PBC mouse model.Among the low,medium,and high doses of ZHLD,the medium dose was the best therapeutic dose to ameliorate the IBEC-EMT of PBC mouse model.