Effect Of Probiotics Combined With ART On Intestinal Mucosal Barrier And Immune Inflammation In HIV Infected Patients: A Preliminary Study

Objective: Since the first discovery of human immunodeficiency virus(HIV)in 1981,it has been raging and popular globally,constitutes a huge challenge to human health,and seriously endangers public safety.Anti-retroviral therapy(ART)has significantly reduced the morbidity and mortality of acquired immune deficiency syndrome(AIDS),which is expected to become a controllable chronic disease.The intestinal tract is an important reservoir of HIV virus.HIV infection leads to the decrease of beneficial bacteria and the increase of pathogenic bacteria in the intestinal tract,which leads to the damage of intestinal mucosal barrier and the continuous immune inflammatory response of the body.At the same time,studies have shown that about half of HIV/AIDS patients are accompanied by diarrhea,abdominal pain and other gastrointestinal symptoms,and ART cannot fundamentally reverse the above intestinal physiological and pathological changes,which has become the bottleneck and challenge of HIV/AIDS treatment.Probiotics are commonly used intestinal microecological regulators,which can regulate the imbalance of intestinal flora,play a role in anti-inflammatory,immune regulation and intestinal mucosal barrier maintenance.It has the potential to improve the intestinal symptoms of HIV/AIDS patients and thus positively regulate the disease course of HIV/AIDS.Therefore,this study intends to observe the effects of probiotics combined with ART on intestinal mucosal barrier and immune inflammation in patients with HIV infection complicated with intestinal symptoms,compared with ART alone,in order to provide new ideas for the treatment of the disease.Methods: A prospective cohort design was used to enroll HIV patients with intestinal symptoms according to inclusion/exclusion criteria.After signing the informed consent form,patients filled in food frequency questionnaire 25(FFQ25),gastrointestinal symptom rating scale(GSRS),36-item short form health survey(SF-36)and other questionnaires.After the evaluation by doctors,the patients were divided into control group(ART)and drug group(ART+ probiotics)by a non-randomized control method.The drug group was further divided into bifidum group(ART+ Bifidobacterium triple viable bacteria)and subtilis group(ART+ Bacillus subtilis double viable bacteria)for a course of treatment for 3 months.The difference of clinical efficacy(improvement of intestinal symptoms)between the control group and the drug group was compared,and drug safety was analyzed.Blood samples and colon biopsy samples were collected before and after treatment.Spearman correlation analysis was used to evaluate the related factors affecting gastrointestinal symptoms.Enzyme-linked immunosorbent assay(ELISA)was used to analyze the effects of intestinal mucosal barrier and immune inflammation before and after treatment.Illumina sequencing was used to analyze the intestinal microbial composition and diversity before and after treatment.Western blotting(WB)was used to analyze the expression of intestinal tight junction related proteins before and after treatment.And immunohistochemistry(IHC)was used to analyze the changes of intestinal signaling pathway proteins Toll-like receptor 4(TLR4)and nuclear factor-kappa B(NF-κB)p65 before and after treatment.Results: Compared with the control group,the related symptoms of diarrhea and abdominal pain were significantly improved in the drug group,and the related scores were significantly decreased(p<0.05).At the same time,there were no obvious abnormalities in liver and kidney function and blood routine before and after treatment,and the safety was good.Spearman correlation analysis showed that the severity of gastrointestinal symptoms was significantly correlated with plasma IL-6 level(p<0.05).GSRS score was significantly negatively correlated with SF-36 score(p<0.05),both in physical component summary(PCS)and mental component summary(MCS)scores,suggesting that gastrointestinal symptoms seriously affected the quality of life of HIV patients.ELISA analysis showed that compared with baseline,supplementation of probiotics significantly reduced plasma levels of lipopolysaccharides(LPS),IL-6 and TNF-α(p<0.05).Illumina sequencing results showed that supplementation of probiotics had no significant effects on colon microbial composition at phylum and genus levels and β diversity(p>0.05),while the Shannon index in α diversity decreased(p>0.05).Western blotting results showed that supplementation of probiotics had no significant effect on the expression of tight Occludin(OCLN)transmembrane protein in intestinal tract(p>0.05).And IHC results show that added probiotics down TLR4/NF-κBp65 protein expression,but no statistical significance(p>0.05).Conclusions: Compared with ART alone,probiotics combined with ART can better relieve enteric-related symptoms in HIV-infected patients with good safety.Probiotics supplementation can reduce the levels of plasma intestinal mucosal barrier and immunoinflammatory markers in patients,but the effect of probiotics on the composition and diversity of colonizing microorganisms in patients’ colons remains to be further demonstrated.The improvement of probiotics on patients’ inflammation may be related to the TLR4/NF-κBp65 pathway,but the exact mechanism still needs further study.