The Study On The Influence Of Abnormal Savda Munziq On The Candidate Gene Expression Of The Model Of Hepatocarcinoma Carrying Of Abnormal Savda Rats

Objective:To investigate the influence of the abnormal savda munziq(ASMq)on the expression level of the candidate gene in the rat model of the hepatocarcinoma which carrying abnormal savda.Methods:(1)The male naive Wistar rats were chosen and randomly divided into six groups.A model of hepatocarcinoma carrying abnormal savda was established by using DEN(diethylnitrosamine)according to Uyghur medical theory.Different doses of ASM such as high(6.0 g/kg),medium(3.0 g/kg),low(1.5 g/kg)were used to intervene the model rats for 20 week.The morphological changes of hepatocarcinoma tissue were observed.(2)The candidate genes were expressed through the whole genome expression spectrum.(3)Selected candidate genes,using mRNA-specific primers and quantitative RT-qPCR method,The mRNA expression level of liver tissue in six groups of rats was analyzed.The mRNA expression level of candidate genes in the liver tissues of each group rat model was determined.(4)Immunohistochemistry and Western-blotting method confirmed STAT3,CyclinD1 gene Protein content change and the phosphorylation of STAT3 protein.Results:(1)The morphology of liver tissue shows that cancerogenic rates in the group of hepatocarcinoma which carrying abnormal savda significantly higher than that of control group of hepatocarcinoma.Cancerogenic rates in ASM intervention group of hepatocarcinoma significantly lower than the group of hepatocarcinoma which carrying abnormal savda.(2)Whole genome expression profiling :Compared with the normal control group,the hepatocarcinoma control group 1964 gene were up-reagulated and 359 gene were down-reagulated.Compared with control hepatocarcinoma group,in the group of hepatocarcinoma which carrying abnormal savda,438 genes were up-regulated and 451 genes were down-regulated.Compared with abnormal savda type hepatocarcinoma group,the high dose group of ASMq 251 gene were up-reagulated,and 396 gene were down-reagulated.Same as above the medium dose group of ASMq 417 gene were up-reagulated,and 386 gene were down-reagulated.the lowe dose group of ASMq 201 gene were up-reagulated and 343 genewere up-reagulated.Among the 11 genes from the abnormal savda type hepatocarcinoma group and ASM intervention group,The trend of expression levels is reversed.so selected STAT3(Signal transductors and activator oftranscription-3),CyclinD1(Cell cycle proteinD1),EGLN3(Prolyl hydroxylase domain protein3),EMP1(epithelialm embrane protejn-1),NEFL(Nerve silk protein light chain),and IGFALS(insulin-like growth factor acid-labile subunit)were Six candidate generic genes.(3)Through RT-qPCR analysis,The accuracy and specificity of the results of the above gene chip are verified.(4)Western-blotting and Immunohistochemical results confirmed that STAT3,CyclinD1 protein expression level was consistent with mRNA level,and STAT3 phosphorylation level was lowest in the normal control group,and the highest in the abnormal savda type hepatocarcinoma group.In the ASMq intervention group,as the ASMq dosage increases,the dose dependences are reduced.Conclusion:(1)The pathological section of liver tissue in Rat model showed that abnormal savda may promote the formation of abnormal savda type hepatocarcinoma and ASMq delay or inhibit the occurrence and development of tumors caused by abnormal savda.To achieve curative effect.(2)Abnormal savda type hepatocarcinoma is closely related to the abnormal regulation of 6 genes such as STAT3,CyclinD1,EGLN3,EMP1,NEFL and IGFALS,and the abnormal expression of these genes can be corrected or reversed by ASMq.