Study On The MicroRNA Expression Of Chronic Hepatitis B With The Patten Of Liver Qi Depression And Spleen Deficiency And Spleen-stomach Dampness-heat

ObjectiveStudy the correlation between the syndrome of liver depression and spleen deficiency, spleen-stomach dampness-heat of Chronic Hepatitis B(CHB) and the differential expression of microRNA(miRNA), aiming to verify the preliminary screening results of miRNA gene chip, find the special miRNAs of two syndrome types of CHB and understand the regulation mechanism of the special miRNAs, then providing experimental basis for the typing of syndrome differentiation of CHB, promoting the objectify and normalize of the CHB syndrome diagnosis.MethodsFrom the results of preliminary studies, we screened out 9 miRNAs with statistically significant different expression in syndrome of liver depression and spleen deficiency of CHB(miR-122-5p,miR-1228-3p,miR-191-3p,miR-4433-5p,miR-223-3p,miR-23a-3p, miR-1225-3p,miR-1281,miR-21-5p),7 miRNAs with statistically significant different expression in syndrome of spleen-stomach dampness-heat of CHB(miR-122-5p,miR-1228-3p,miR-191-3p,miR-4433-5p,miR-1280,miR-150-3p,miR-765), four of the miRNAs in the two syndrome types of CHB were the same. Applying Real-time fluorescence quantitative PCR TaqMan probe methods we validated the differential expression of the 12 miRNAs in 10 CHB patients with syndrome of liver depression and spleen deficiency,10 CHB patients with syndrome of spleen-stomach dampness-heat and 3 healthy people. All data were analyzed with the spss20.0 software.Results1. Compared with control group, the expression of miR-122-5p and miR-1281 of the liver depression and spleen deficiency group were up-regulated, while the expression of miR-23a-3p was down-regulated, the differences had statistical significance (P<0.05); the expression of miR-1228-3p and miR-1225-3p were up-regulated, while the expression of miR-223-3p and miR-191-3p were down-regulated, the differences had no statistical significance (P>0.05).2. Compared with control group, the expression of miR-122-5p in spleen-stomach dampess-heat group was up-regulated, while the expression of miR-1280 was down-regulated, the differences had statistical significance (P<0.05); the expression of miR-1228-3p, miR-191-3p and miR-150-3p were down-regulated, the differences had no statistical significance (P>0.05).3. Compared with the spleen-stomach dampness-heat group, the expression of miR-122-5p, miR-1228-3p and miR-191-3p of the liver depression and spleen deficiency group were up-regulated,and the differences had no statistical significance (P>0.05).Conclusions1. This study showed that there existed 2 special miRNAs in the plasma of CHB patients with syndrome of the liver depression and spleen deficiency:miR-1281 and miR-23a-3p. The results were as the same as the miRNA gene chip’s, which participated in the regulation of cell proliferation, immunoregulation, inflammatory reaction, NK cell viability and other process through the targeted combination with the target gene, and had certain correlation with the CHB with syndrome of the liver depression and spleen deficiency, which showed that they could be regarded as the biological markers in the diagnosis of CHB with syndrome of the liver depression and spleen deficiency.2. This study showed that there existed 1 special miRNA in the plasma of CHB patients with syndrome of the spleen-stomach dampness-heat:miR-1280, which was as the same as the result of miRNA gene chips, which participated in the immune responseinflammatory injury, protein metabolism and other process through the targeted combination with the target gene, and had certain correlation with the CHB with syndrome of the spleen-stomach dampness-heat, which showed that it could be regarded as the biological markers in the diagnosis of CHB with syndrome of the spleen-stomach dampness-heat.3. This study showed that there existed 1 special miRNA in the plasma of CHB patients: miR-122-5p. The result was as the same as the miRNA gene chip’s, it participated in the regulation of cell proliferation, HBV virus infection, inflammatory injury of liver, hepatic fibrogenesis and other process through the targeted combination with the target gene, and had certain correlation with the CHB, which showed that it could be regarded as the biological markers in the diagnosis of CHB.