Study Of The Relationship Between The Levels Of Hepatocyte Nuclear Factor 4 Alpha And 3 Beta And HBV Replication In The Liver Of HBV Infected Patients

Background and Aims Hepatocyte nuclear factor 4 alpha (HNF-4α) and HNF-3βare the members of liver-enriched transcription factor (LETFs) which play important roles in regulation of HBV replication. With cell culture and animal model systems, we showed that HNF-4αcan support HBV replication in nonhepatoma cells and HNF-3βinhibits HBV replication. But the expression of HNF-4αand HNF-3βin the liver tissues of chronic HBV infected patients and the relationship between the level of HNF-4αand HNF-3βand HBV replication are not yet clear. The aim of this study is to investigate the expression levels of HNF-4αand HNF-3βand their relationship with the level of HBV replication in the liver biopsy tissues of chronic HBV infected patients.Methods Liver biopsy specimens from 86 chronic HBV infected patients were collected. Some tissues were sliced and mounted to the same glass supports, the others were made into ordinary slices. Using immunohistochemical staining method, the expression levels of HNF-4α,HNF-3βand HBsAg, HBcAg in the serial sections of liver biopsy tissues were checked. Preparing Dig labeled HBVDNA probes, and the levels of HBVDNA in the serial sections of liver biopsy tissues were detected by in situ hybridization technique.On the basis of Axiotis score standard, our assessment is according to both the percentage of positive cells and the staining intensity. The proportion of positive cells accounted for total similar cells score criteria: positive cells 0%～10% (0), 11%～25%(1), 26%～50%(2), 51%～75%(3), 76%～100%(4); Cell staining intensity score: 0 for no color, yellow or blue for 1, brown or purple blue to 2, tan or blue-black to 3. The sum of the two scores shall be the final score we need.On the results of Kappa consistency test, correlation analysis and t tests, all statistics analysis are completed through the SPSS statistical software package. The relationship between the levels of HNF-4αor HNF-3βexpression and HBV replication was also been compared and analyzed.Results1. The expression levels of HNF-4α,HNF-3βand HBsAg, HBcAg, HBVDNA in 86 Liver biopsy specimens of chronic HBV infected patientsExperimental results displayed that the expression of HNF-4α,HNF-3βand HBVDNA were cell nucleus / cytoplasm positive, the expression of HBsAg was cytoplasm positive, and the expression of HBcAg is cell nucleus / cytoplasm / cell membrane positive. According to Axiotis score standards, the scores of above markers are shown in table 1. Table 1. The expression levels of HNF-4α,HNF-3βand HBsAg,HBcAg,HB VDNA The scores range in table 1 is from "0" to "7", the sum of above two scores. "2.5" represents score of 2～3, so on and so forth.2. The statistical analysis of the scoresStatistical linear correlation analysis shows that in the samples with higher levels of HNF-4α, the levels of HBsAg, HBcAg and HBV DNA are also higher, there is a positive correlation between the expression level of HBsAg and HNF-4αHBcAg and HNF-4αand the level of HBV DNA and HNF-4α(P＜0.01, R＞0); In contrast, in the samples with higher levels of HNF-3βexpression, the levels of HBsAg, HBcAg and HBVDNA are relatively lower. Statistical analysis showed that there is a negative correlation between the expression level of HBsAg and HNF-3β, HBcAg and HNF-3βand the level ofHBV DNA and HNF-3β(P＜0.01, R＜0).Table 2. the statistical analysis of the expression of HNF-4α,HNF-3βand HBsAg, HBcAg,HBVDNAIn table 2, "r" is for the correlation coefficient, its value range is -1～1. r＞0 represents the positive correlation; r＜0 represents the negative correlation; "v' represents freedom, v=84 in this experiment, then we check the "t value sector table" and observe the P value. "P＜0.01" shows there is statistical significance between the two variables.Conclusion The result of this study indicates that there is a correlation between the expression level of HNF-4αand HNF-3βand HBV reproduction in chronic HBV infected patients. There is a positive and negative correlation between HNF-4αand HNF-3βexpression with HBV replication, respectively, in the liver of chronic HBV infected patients. This suggests that HNF-4αand HNF-3βare likely to participate in the process of HBV replication in patients with HBV infection. HNF-4αmay support HBV replication, while HNF-3βmay inhibit HBV replication. This will provide the theoretical basis for deeply studying on human HBV replication mechanism and guiding clinical practice in future .However, the detailed relationship of HNF-4αand HNF-3βexpression with HBV replication and which is in a dominant position, are still need in-depth study and research.