Study On The Relationship Between Notch With Wnt Signaling Pathways And Hepatic Fibrosis Markers

Objective:Liver fibrosis is a common wound healing response in chronic liver injury,which can be caused by a variety of causes,including alcohol or drug toxicity,persistent viral infection,genetic factors and so on.The key of this process is the activation of hepatic stellate cells which characterized by expressing alpha smooth muscle actin,secreting type I collagen and other components of the extracellular matrix.The Notch and Wnt signaling pathways are widely distributed in organisms,which can regulate the cell growth,proliferation,differentiation,apoptosis and other physiological processes.Mounts of results show that these two pathways may be closely related to the formation of liver fibrosis.This study aim to explore the influence of the above pathways on liver fibrosis through the establishing liver fibrosis model,detecting the mRNA expression of key factors in Notch and Wnt signaling pathways.Methods:1.Building model:20 healthy male Sprague Dawley rats were in abrosia before the experiment,randomly divided into normal control group(n=10)and liver fibrosis model group(n=10).40% carbon tetrachloride was injected into subcutaneous of rats which the model group(0.6 mL/100 g body weight,the first double agent).While the normal control group was instead of 0.9%NaCl injected with the same method every three days.This period persists of eighth weeks.Following sacrifice of rats,the liver was immediately placed in the low temperature environment.2.Index detection : RNA was extracted from two groups of liver specimens,and cDNA was synthesized by reverse transcription reaction.Quant Reverse Transcriptase chain reaction was used to detect mRNA expression of ?-SMA,COL1 ?2,Notch3,Wnt5 a,?-catenin and GAPDH in both groups.3.Statistical analysis:The differences between Notch and Wnt signaling molecules in the two groups were compared with two independent samples t test,and the correlation between Notch and Wnt signaling pathway was analyzed with Persion correlation analysis method.Analysis of all experimental data using SPSS22.0 software.The difference was statistically significant with P<0.05.Results:The expression of m RNA in liver tissue of rats with hepatic fibrosis index and Notch,Wnt signaling molecules: Q-PCR test results showed that compared with normal control group,expression of ?-SMA,COL1?2,Notch3,Wnt5 a was increased,the difference was statistically significant(P<0.05),expression of ?-catenin had no significant difference(P>0.05).Among the indexes,?-SMA was positively correlated with Wnt5a(r= 0.689,P<0.05),COL1?2 was positively correlated with Notch3(r=0.676,P<0.05).Conclusion:Notch and Wnt signaling pathways are involved in the occurrence and development of hepatic fibrosis.Selective intervention of Notch3 and Wnt5 a transduction may be a new approach to anti hepatic fibrosis.