MiR-133a Effect Study On Echinococcus Multilocularis Induced Hepatic Stellate Cell Activation In Vitro And Liver Fibrosis In Infected Mice In Vivo

Objective: To study the miR-133 a effect on echinococcus multilocularis(Em) induced hepatic stellate cell activation in vitro and liver fibrosis in infected mice in vivo. Methods: Mice were injected by Em and control group were injected with saline. Mouse liver tissues were isolated and detected after 1 month and 3 months Em Infection. The pathological results were determined by H&E staining and the expression of Type I collagen was proved by Sirius scarlet trinitrophenol staining. Alpha smooth muscle actin(a-SMA) expression was detected by immunohistochemical staining. The mRNA expressions of a-SMA, COL1A1 and miR-133 a were analyzed by Reverse Transcription quantitative PCR(RT-qPCR). LX-2 cells were stimulated by Em protein(60μg/ml and 600μg/ml) and the expressions of α-SMA, COL1A1, Smad2,Smad3,Smad4,TGFβ-RI,TGFβ-RII and miR-133 a were detected by RT-qPCR and Western blotting. miR-133 a overexpression was detected in miRNA-133 a mimics transfected LX-2 cell and then the mRNA and protein expressions of α-SMA, COL1A1,Smad2,Smad3 and Smad4 were dected by RT-qPCR and Western blotting. Results:(1) Echinococcus multilocularis infected mouse model was established successfully. Compare to sham group, the mRNA and protein expressions of a-SMA and Type I collagen were significant increased in the liver of Em infected mice(P<0.01), which close to the lesion, after Em infected 1 month or 3 months, meanwhile, the miR-133 a expression was significant increased in the liver of Em infected mice(P<0.01), which close to the lesion, after Em infected 1 month.(2) Isolatied Em protein could significant improved α-SMA, Type I collagen and miR-133 a expression in Em protein pretreated hepatic stellate cells(HSCs)(P<0.0001). Transfected mi RNA-133 a mimics could significant improved the expressions of α-SMA, COL1A1, Smad2, Smad3 and Smad4 in Em protein pretreated LX-2 cells(P<0.05). Conclusion: miR-133 a may play a role on HSC activation through TGF-β/Smad pathway.