Studies On The Relationship Between Serum Levels Of 25（OH）D₃ And Chronic Hepatitis B Virus Infection

[Background and Objectives]Chronic hepatitis B virus(HBV)infection refers to a history of hepatitis B lasting 6 months or more or hepatitis B surface antigen(HBsAg)positive for up to 6 months or more,and now HBsAg and or HBV DNA is still positive.According to chronic HBV infection history,symptoms and signs,liver function,biochemical index,blood coagulation index,virological indicators and imaging and pathological examinations,chronic HBV infection is usually clasified into five types:asymptomatic carriers(ASC),occult hepatitis B virus infection(OBI),chronic hepatitis B(CHB),liver cirrhosis(LC),and hepatocellular carcinoma(HCC).Vitamin D(Vit D)is an essential nutrient for human being,belonging to the steroid derivatives.Vitamin D has many biological activities,the most well-known being the regulation of calcium and phosphorus metabolism,and adjusting metabolism changes in bone.Vitamin D participates in immune regulation,differentiation and proliferation of cells,plays an important role in anti-infection,anti-fibrosis,anti-oxidation,anti-tumor and other aspects,and has influence on prevention and treatment of cardiovascular,endocrine and immune diseases.In recent years,more and more researchers found that vitamin D plays an important role in chronic liver disease(CLD),such as alcoholic hepatitis,fatty liver,HBV infection related liver disease,HCV infection related liver disease,autoimmune hepatitis.When liver dysfunction occurs,25-hydroxylase activity in liver will reduce and synthesis of Vit D will be hindered since the formation of Vit D needs hydroxylation of liver enzyme.Meanwhile absorption of Vit D will also reduce.Although diagnosis and treatment of chronic HBV infection has made great progress in recent 20 years,interferon-αand other immune modulators only have a very low efficacy for chronic hepatitis B virus infection,hepatitis B is quite easy to relapse after short term use of nucleoside(acid)analogues(NAs)and long-term use of NAs increases drug resistance.Furthermore,there is no way to overcome drug resistance so far.Therefore,new therapies for chronic HBV infection and new prodictors for judgement of patient’s situation are important for stabilities of the disease,improvement of virus response and enhancement of quality of life.This research aims at analysis of serum levels of 25(OH)D3 in patients with chronic HBV infection,exploration of the relationship between serum level of vitamin D3 and liver function,HBV serological markers,virological makers,liver disease progression,and confirmation of clinical significance of 25(OH)D3 in chronic HBV infection.[Methods]The subjects consist of 99 healthy control(HC)and 369 patients with chronic HBV infection diagnosed in Jinan Infectious Disease Hospital from January 2015 to August 2016,including asymptomatic carriers(ASC)90 cases,chronic hepatitis B(CHB)116 cases,liver cirrhosis(LC)111 cases,and primary hepatocellular carcinoma(HCC)52 cases.Their demographic data,liver function,hepatitis B virus serological and virological examination results and their sera were all collected.ELISA kits were used to detect the content of 25-hydroxy vitamin D3(25(OH)D3)in serum.Data were analyzed with SPSS 18.0 statistical software.[Results]As a result,the level of serum 25(OH)D3 in 99 healthy control(HC)was 32.99± 7.33 ng/ml,and significantly higher than that of 369 patients with chronic HBV infection(29.28 ± 7.33 ng/ml)with a statistically significant difference(P<0.001).The average contents of serum 25(OH)D3 in sera of 90 cases with asymptomatic HBV carriers(ASC),of 116 cases with chronic hepatitis B(CHB),of 111 cases with liver cirrhosis(LC)and of 52 cases with hepatocellular carcinoma(HCC)were 26.88 ± 75.88 ng/ml,31.49 ± 8.04 ng/ml,28.85 ± 6.80 ng/ml and 29.38 ±7.77 ng/ml respectively.The results showed that the serum level of serum 25(OH)D3 was lowest in ASC group,and followed by LC group and HCC group.Vitamin D in CHB group reduced the least.The contents of serum 25(OH)D3 in various clinical stages of chronic HBV infection were all significantly lower than that in HC group(ASC,P<0.001;CHB,P=0.27 group;LC,P<0.001;HCC,P=0.006),except for CHB group.The content in ASC group was statistically and significantly lower than that of CHB,LC and HCC groups(CHB,P=0.000;LC,P=0.029;HCC,P=0.048).In addition,there was statistically significant difference between LC group and CHB group(28.85 ± 6.80 ng/ml vs 31.49 ± 8.04 ng/ml,P=0.008),but there was no significant difference between LC group and HCC group(28.85 ± 6.80 vs 29.38 ±7.77 ng/ml,P=0.679).The decrease of serum 25(OH)D3in patients with chronic HBV infection is more serious than in healthy control group(HC).There were 209 cases(56.64%)whose serum 25(OH)D3 was not sufficient(≤29.9ng/mL)in chronic HBV infection,and 24 cases(24.24%)in healthy control group.Statistically significant difference exist between them(P<0.001).Those with insufficient level of serum 25(OH)D3(20-29.9ng/mL)and those with deficiency of serum 25(OH)D3(<19.9ng/mL)in ASC group were 50 cases(55.56%)and 15 cases(16.66%),in CHB group 35 cases(30.17%)and 13 cases(11.21%),in LC group 59 cases(53.15%)and 9 cases(8.11%),and in HCC group 24 cases(46.15%)and 4 cases(7.7%)respectively.The percentages of those with insufficient serum 25(OH)D3(20-29.9ng/mL)in ASC(55.56%),CHB(30.17%),LC(53.15%)and HCC group(46.15%)were all significantly higher than in HC group(20.2%,P=0.000,0.044,0.000,and 0.000 respetively).Spearman correlation analysis found that serum 25(OH)D3 levels in ASC group were negatively correlated with gender(r=-0.24,P=0.024),HBeAg(r=-0.49,P=0.000)and HBV DNA classification(r=-0.52,P=0.000).Female,HBeAg negative and low HBV load had higher serum levels of vitamin D3.Nevertheless,serum content of serum 25(OH)D3 had no correlationship with age,ALT,AST,ALB,HBsAg,and HBcAb.A multivariate linear model analysis with all the relevant indicators demonstrated that only HBV DNA load was an independent factor for serum 25(OH)D3(r=-2.82,P=0.003).In order to further study the influencial factors of serum 25(OH)D3,all the indicators of gender,age,ALT,AST,ALB,HBsAg classification,HBcAb classification,HBV DNA classification of the 369 patients with chronic HBV infection were included in the analysis,and the results showed that the content of serum 25(OH)D3 was only negatively correlated with HBV DNA(r=-0.153,P=0.003)and not correlated with other factors.The high viral load group had the lowest content of 25(OH)D3(27.39±7.20 ng/mL),and the low viral load group had the highest content(30.29±6.76 ng/mL).There was a significantly statistical difference(P=0.013)between them.Those with deficiency of serum 25(OH)D3(less than 19.9ng/mL)in the middle viral load group accounted for 17.93%and in the high viral load group accounted for 13.73%,both were significantly higher than those in the low viral load group(4.63%,),P<0.01).[Conclusion]1.This study showed that the decrease of serum 25(OH)D3 was common in patients with chronic HBV infection;the serum levels of 25(OH)D3 in various clinical stages were all lower than in healthful controls,the asymptomatic carriers had the lowest level of vitamin D3,and the highest incidence of vitamin D deficiency.2.The levels of serum 25(OH)D3 in patients with liver cirrhosis and hepatocellular carcinoma were significantly lower than that in patients with chronic hepatitis B,and the magnitude of the decline in patients with cirrhosis was seemingly more than that in patients with hepatocellular carcinoma;3.Hepatitis B virus load was negatively correlated with serum level of 25(OH)D3,and an independent factor for vitamin D3;This research demonstrated that the decrease of vitamin D was more obvious in chronic HBV carriers,a high proportion of patients with vitamin D deficiency in such a population,whether there was a similar phenomenon in the chronic HBV carriers in other parts of China and elsewhere,requires large-scale epidemiological investigations to confirm.The results suggest that we should pay attention to HBV carriers about vitamin D deficiency in clinical practice.The level of vitamin D deficiency in patients with liver cirrhosis was higher than that in patients with hepatocellular carcinoma;did it suggest that HCC cells have the ability to synthesize vitamin D in some way was worthy of further study.The content of serum 25(OH)D3 was negatively correlated with HBV load,indicating that maintaining a certain level of 25(OH)D3 may promote the efficacy of antiviral therapy.