The Relationship Between The Expression Levels Of MANF Protein And The Degree Of Liver Inflammation And Fibrosis

Objective To explore the role of MANF protein in the development of liver inflammation and fibrosis, and analyse the relationship between the level of MANF protein and clinical characteristics in the patients with chronic HBV infection. Methods The 20 cases liver tissues of chronic HBV infection patients in different stages were selected to investigate the expression and distribution of MANF protein, and analyse the relationship between its expression and the degree of liver inflammation and fibrosis by isobaric tags for relative and absolute quantitation(ITRAQ) and immunohistochemistry. Real time polymerase chain reaction(PCR) were applied to detect the MANF m RNA level in peripheral blood mononuclear cells(PBMCs) of patients from liver cirrhosis(LC), chronic hepatitis B(CHB), asymptomatic hepatitis B virus carriers(ASC) after chronic HBV infection and normal healthy controls(HC). Spearman rank correlation analysis was used to analyse the relationship between the level of MANF m RNA and liver disease progression, and analyzed its correlation with HBV loads, hepatitis B virus surface antigen( HBs Ag) quantitative, hepatitis B virus e antigen(HBe Ag) qualitative, transaminase level, total bilirubin( TBIL) level and the clinical pathological physiological factors. Results 1. MANF protein was mainly expressed in the cytoplasm of liver cells. The results showed more negative and weakly positive expression in G1S1 group, more weakly positive and positive expression in G2S2 group and more positive and strongly positive expression in G3S3 and G4S4 groups. The expression of MANF protein was gradually increased with the exacerbation of liver inflammation and fibrosis. 2. The MANF m RNA level in PBMCs of differentdiagnosis groups were compared, its level is not the same among four groups(c2 =28.171,P = 0.00), its level in NC, ASC and CHB group were all lower than that in LC group(Z=-4.182/-4.313/-2.842, P <0.01). The rest of the differences between any two groups had no statistical significance. 3. The expression levels of MANF m RNA in PBMCs were gradually increased from HC to LC. There was a positive correlation between the expression of MANF m RNA and the progression of liver disease(rs=0.376,P=0.00). 4. The MANF m RNA levels in the PBMCs of different clinical indicators groups were compared. There is a statistically significant difference among three subgroups(HBs Ag<1.5×106IU/L, 1.5×106 IU/L- 2.0×107IU/L and > 2.0×107IU/L, c2 =6.297, P <0.05). The MANF m RNA expression level of HBe Ag positive group was higher than negative group(Z=-3.265, P < 0.01).The MANF m RNA expression level of normal TBIL group was lower than the abnormal group(Z=-3.446, P < 0.01). Conclusion 1. Compared with NC, there is a rising trend of MANF m RNA expression level in the PBMCs of ASC, CHB and LC, especially in LC patients. While MANF gene is a selected gene from 30000 genes and also the most sensitive one to endoplasmic reticulum(ER) stress, therefore, this further validates the existence of ER stress in patients with chronic HBV infection. The MANF expression was positively correlated with disease progression after chronic HBV infection, suggesting that ER stress may be involved in the disease progression of chronic HBV infection. 2. After chronic HBV infection, the expression of MANF protein was gradually increased with the exacerbation of liver inflammation and fibrosis, there was a positive correlation between the expression of MANF m RNA in PBMCs and the progression of liver disease. It indicates that MANF may participate in the progress of liver inflammation and fibrosis after chronic HBV infection.