| ObjectiveThe objectives of this study are to investigate the changes of the fecal HBV DNA,ratio between the intestinal flora and IL-17A~F in patients with CHB,as well as test the influence of intestinal probiotics treatment on these parameters.On the basis of the above results and the change of liver function,we want to clarify the therapeutic effect and possible toxic effects of intestinal probiotics in the treatment of chronic hepatitis B.Materials and Methods66 patients with CHB were divided into two groups in random: the conventional treatment group(antiviral therapy combined with liver protection,33 patients)and the micro ecological treatment group(conventional therapy combined with Clostridium butyricum double live germs,33 patients).The stools and serum of 66 patients with CHB and 30 healthy volunteers were collected before treatment and after 13±1 day treatment.HBV DNA and intestinal flora DNA were detected by real-time PCR.Interleukin(IL)-17A~F,IL-1?,IL-6,and CXCL-13 in serum were detected by ELISA,and liver function was also analyzed.The ratio of intestinal flora was calculated.Before treatment,the data and relationships among HBV DNA,ratio between the intestinal flora,liver function and IL-17A~F were analyzed.The data were compared among healthy volunteers,CHB before and after treatment.Moreover,the difference between the conventional treatment group and micro ecological treatment group were compared.Result one1.The copy numbers of HBV DNA from the stool and serum of 66 patients were all above the lower detection limit(0.5 x 103 IU/ml)and were positively correlated(r=0.57).HBV DNA copy numbers of HBeAg-positive group were higher than HBeAg-negative group(P=0.015).Fecal HBV DNA was negatively correlated with ALP(r=-0.41)and TBIL(r=-0.29).There were no difference in the negative conversion rate of HBV DNA between in serum and in stool after antiviral treament(P>0.05).The liver function improved obviously in the negative conversion group of fecal HBV DNA and serum HBV DNA(P<0.05),and there were no difference in two groups after antiviral treament(P>0.05).2.Compared to healthy volunteers,the copy numbers of Enterobacteriaceae,Enterococcus Prevotella,Clostridium,Candida albicans and microflora ratio of EB/BI,EB/L,EB/R,EC/BI,EC/L,EC/R,C/BI,C/L,C/R,CA/BI,CA/L,CA/R,BR/BI,BR/L,BR/R,EB/P,EC/CA,EC/BR,EC/P,C/CA,C/BR,C/P,CA/P in CHB patients were significantly increased.However,the copy number of Bifidobacterium,Lactobacillus,Prevotella,Ruminococcus and microflora ratio of EB/EC,EB/C,EC/C in CHB patients were significantly decreased compared to healthy volunteers,and there were no changes of EB/CA,EB/BR,EC/BR,CA/BR,BI/L,L/R and BI/R(P>0.05).3.Fecal HBV DNA was positively correlated with Enterococcus,EC/CA,EC/P,EC/R(P<0.05)and was negatively correlated with EB/EC.Serum HBV DNA was positively correlated with EC/L,CA/L,P/L(P<0.05)and was negatively correlated with Lactobacillus,L/BI(P<0.05).4.Before treatment,there were correlation among the intestinal flora,ratio between the intestinal flora and the changes of liver function in 66 patients with CHB.TBIL was positively correlated with Enterobacteriaceae,Candida albicans,Ruminococcus,EB/BI,EB/C,EB/BR,EC/BI,C/BI,CA/BI and CA/P,and was negatively correlated with Bifidobacterium and C/R.Especially,TBIL was positively correlated with EB/P and R/BI,and the correlation was the strongest(r=0.42).TBIL was negatively correlated with BR/R and P/R,and the correlation was the strongest(r=-0.34).ALB was positively correlated with EB/R,L/BI,C/R and P/R,and was negatively correlated with Candida albicans,Ruminococcus and BR/B.Especially,ALB was positively correlated with CA/R,and the correlation was the strongest(r=0.40).ALB was negatively correlated with R/BI,and the correlation was the strongest(r=-0.34).ALT was positively correlated with Enterobacteriaceae,Lactobacillus,Clostridium,EB/CA,EB/BR,EB/P,EC/BR,EC/P,L/BI,C/P,C/BR,CA/BR and CA/P,and was negatively correlated with CA/L,BR/L,P/L,C/R,CA/R and BR/R.Especially,ALT was positively correlated with Ruminococcus,and the correlation was the strongest(r=0.47).ALT was negatively correlated with P/R,and the correlation was the strongest(r=-0.43).AST was positively correlated with Enterococcus,Candida albicans,EB/BI,C/BI,CA/BI and R/BI,and was negatively correlated with Bifidobacterium,Bacteroides,Prevotella,P/L,BR/R and P/R.Especially,AST was positively correlated with EB/BR,EB/P,EC/BI,EC/BR,EC/P,L/BI,C/BR,C/P,CA/BR and CA/P,and the correlation was the strongest(r>0.6).AST was negatively correlated with BR/L and BR/P,and the correlation was the strongest(r=-0.56).ALP was positively correlated with EB/BI,EB/BR,EC/BR,C/BI,CA/BI,P/BI,R/BI,C/P and CA/BR,and was negatively correlated with Bacteroides and BR/R(P<0.05).Especially,ALP was positively correlated with EC/BI,and the correlation was the strongest(r=0.51).ALP was negatively correlated with Bifidobacterium,and the correlation coefficient was the strongest(r=-0.56).GGT was positively correlated with Enterococcu,Lactobacillus,EC/BI and L/BI,and was negatively correlated with CA/L and P/L.Especially,GGT was positively correlated with Clostridium,and the correlation was the strongest(r=0.37).GGT was negatively correlated with BR/L,and the correlation was the strongest(r=-0.28).5.The expression of serum IL-17A~F in patients with CHB before treatment were significantly higher than those in healthy volunteers,and were significant significantly higher than those in patients after treatment.6.Before treatment,fecal HBV DNA was positively correlated with IL-17 B,IL-17 D and IL-17 E.Serum HBV DNA had no correlation with IL-17A~F.7.Before treatment,IL-17 A was positively correlated with EC/CA and EC/R,IL-17 B was negatively correlated with R/L,IL-17 C was negaitively correlated with EB/EC,BR/L and P/L.IL-1D was positively correlated with EC/BI.8.Before treatment,IL-17A~F were positively correlated with IL-1?,IL-6 and CXCL-13(r>0.7).9.Before treatment,IL-17A~F were positively correlated with TBIL and DBIL.IL-17 C was positively correlated with TBA.IL-17 B and IL-17 E were positively correlated with ALB.Result two1.The negative conversion rate of serum HBV DNA in the micro ecological treatment group and conventional treatment group was 33.30%and 24.24%,respectively.And there was no difference between two groups(P>0.05).The negative conversion rate of stool HBV DNA in the micro ecological treatment group was 15.15%,and 30.30% in the conventional treatment group,and there was no difference between two groups(P>0.05).2.Patients in the micro ecological treatment group exhibited significantly decreased Enterobacteriaceae,Bifidobacterium,ratio of EB/EC,EB/C,EB/CA,EB/BR,EB/P and EB/R(P<0.05),and significantly increased Enterococcus,ratio of EC/C,EC/CA,EC/BI,EC/L,EC/R,C/BI,CA/BI and P/BI compared to patients in the conventional treatment group(P<0.05).3.Patients in the micro ecological treatment group exhibited significantly decreased IL-17 A,IL-17 C,IL-17 D and IL-17 F compared to patients in the conventional treatment group(P<0.01).4.Patients in the micro ecological treatment group exhibited significantly improved ALT,AST and GGT,but significantly increased TBIL compared to patients in the conventional treatment group(P<0.05).Conclusion1.Our observations suggest that HBV DNA can be detected in the stool of in patients with CHB and fecal HBV DNA and serum HBV DNA might have different clinical significance.2.There are a variety of disorder of ratio between the intestinal flora and changes of IL-17A~F in patients with CHB.The changes of IL-17A~F are associated with the disorder of ratio between the intestinal flora,and they are all associated with the liver damage in Patients with CHB.3.The treatment of Clostridium butyricum double live germs can promote the repair of liver cell damage.The mechanism may be related to improve the degree of disorder of the ratio between the intestinal flora and decrease expression of IL-17.However,the treatment of Clostridium butyricum double live germs can bring the new disorder of the ratio between the intestinal flora and the new liver damage. |
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