Effect Of The Intensive Phase Of Anti-tuberculosis Treatment On Liver Function And The Improvement Effect Of Probiotic Supplementation

Objective:During two months of intensive treatment,patients with pulmonary tuberculosis need to take anti-tuberculosis drugs in large dosage and multiple forms.These drugs can effectively inhibit or kill Mycobacterium tuberculosi,however,they can also cause liver dysfunction or liver injury.Probiotics have been considered as beneficial microecological modulators in gastrointestinal function.With the widespread application of gut-liver axis theory,several studies have shown that certain probiotics can produce beneficial effects in liver diseases,but it has not been reported whether probiotics supplementation has effects on liver dysfunction or injury caused by anti-tuberculosis drugs.The purpose of this study is to investigate the incidence of liver dysfunction in tuberculosis patients during intensive treatment.On this basis,the probiotics supplementation trial was carried out to observe the effects on liver function,intestinal permeability,inflammation levels,and bile acid during intensive treatment.Methods:From January 2011 to December 2019,1,580 tuberculosis patients aged 18 years or older who met the inclusion and exclusion criteria were selected in a chest hospital in Shandong Province.The basic information and clinical information of the subjects were collected,and the incidence rate of liver dysfunction during intensive treatment was analyzed.The dietary intake of the subjects was investigated by SFFQ,and the association between dietary intake and liver dysfunction was analyzed.The clinical trial was conducted,and 325 tuberculosis patients aged 18 years or older were included.Participants were simple-randomly allocated to three groups:the low-dosage L.casei group(1×10¹⁰colony-forming units[CFU]daily);the high-dosage L.casei group(2×10¹⁰CFU daily);the control group（without L.casei intervention）.The supplementary intervention period of L.casei lasted for two months.The basic information and clinical information were collected.The blood and stool samples were collected two months after the trial,and were frozen in a-80℃ultra-low temperature freezer in time for later indexes detection.Serum alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,total bilirubin（TBi L）,direct bilirubin（DBi L）,and indirect bilirubin（IBi L）,Alkaline phosphatase（ALP）,γ-glutamyl transferase（GGT）were detected by automatic biochemical analyzer.Plasma lipopolysaccharide（LPS）,zona occludens 1（ZO-1）,intestinal fatty acid binding protein（IFABP）,tumor necrosis factor-ɑ（TNF-ɑ）,white blood cells Interleukin 6（IL-6）and Interleukin 12（IL-12）were detected by enzyme-linked immunosorbent assay（ELISA）.Plasma metabolites were detected by liquid chromatography-mass spectrometry（LC-MS）.Under the same chromatographic conditions,the retention time and mass-to-charge ratio of the bile acid standards were compared with the retention time and mass-to-charge ratio of the chromatographic peak in the plasma samples,and the peak area indicated the levels of cholic acid（CA）,chenodeoxycholic acid（CDCA）,deoxycholic acid（DCA）,and lithocholic acid（LCA）.Plasma metabolites between the two groups were screened by volcano map,based on the fold change value and P value.The effects of on liver function,intestinal function,and metabolites were analyzed by Analysis of Variance,Kruskal-Wallis test,and Bonferroni test and other statistical methods.Results:In this study,the overall incidence of liver dysfunction in 1,580 tuberculosis patients was 25.82%during intensive treatment,which is more common in young people,male and mild dysfunction.Dietary intake survey indicated that decreased vegetables intake was associated with increased risk of liver dysfunction during intensive treatment.Compared to the highest tertile（>200.0g/d）,the OR of the lowest tertile（<78.6g/d）of vegetable intake,the OR of the middle tertile（78.6-200.0g/d）of vegetable intake and the risk of liver dysfunction was 2.10（95%CI:1.11-3.95）,2.37（95%CI:1.31-4.29）,respectively.After two months of intervention with L.casei,the overall incidence of liver injury in 325 tuberculosis patients was 4.6%.The incidence of liver injury in the control group,the low-dosage L.casei group and the high-dosage L.casei group were 5.7%,4.7%,and 3.5%,respectively（P>0.05）.The results of liver function in blood showed that compared with the control group,the incidence of elevated ALP in the low-dosage L.casei group was also slightly lower（3.5%vs.4.9%,P=0.092）,and the incidence of elevated ALP in the high-dosage L.casei group was significantly lower（0 vs.4.9%,P=0.024）.Compared with the control group,the low-dosage L.casei group had a lower incidence of elevated TBi L（1.2%vs.9.7%,P=0.013）and IBi L（21.2%vs.35.0%,P=0.038）.The results of intestinal permeability showed that the level of LPS,ZO-1 and IFABP in the high-dosage L.casei group were significantly lower than those in the control group and the low-dosage L.casei group（P<0.05）.The results of inflammatory factors showed that the levels of TNF-ɑand IL-12 in the high-dosage L.casei group were significantly lower than that in the control group and the low-dosage L.casei group（P<0.05）.Compared with the control group,the level of IL-6 in the low-dosage and high-dosage L.casei group was lower（P<0.05）.The results of bile acid metabolism showed that after two months of intervention,probiotic supplementation had no significant effect on the levels of CA,CDCA,DCA,and LCA.The results of differential metabolite showed that compared with the control group,the levels of retinoic acid,phosphatidylethanolamine,phosphatidylcholine,linolenic acid,11,12-eicosatetraenoic acid were lower in the high-dosage L.casei group（P<0.05）.Compared with the low-dosage L.casei group,the levels of retinoic acid,phosphatidylethanolamine,phosphatidylcholine,11,12-epoxyeicosatrienoic acid were lower,the level of phosphatidylserine was higher in the high-dosage L.casei group（P<0.05）.Conclusion:Tuberculosis patients are prone to liver dysfunction during intensive treatment,which is more common in young people,male and mild dysfunction.A high vegetable intake was associated with a low risk of liver dysfunction during tuberculosis treatment.L.casei supplementation can suppress abnormally elevated cholestasis related liver indices,which may be related to the reduction of intestinal permeability,inflammation level and the regulation of retinol metabolism,lipid metabolism.It is recommended that dietary adjustment and appropriate L.casei supplementation during treatment should be adopted to alleviate the adverse effects of liver dysfunction and intestinal dysfunction,and improve the tolerance of tuberculosis patients.