| BackgroundSystemic lupus erythematosus(SLE)is a kind of systemic autoimmune disease with the clinical features of influencing multiple systems and organs,constant relapse and abnormal autoimmunity.If it is not treated timely,it will cause irreversible damages to influenced organs and finally lead to death.The pathogenesis of SLE is very complicated and not mastered completely yet.It is believed universally that the disease is related to genes,environmental factors and estrogen level.With the rapid development of microbial community and metabolome analysis and other high-throughput sequencing technologies,more and more researches show that SLE patients and mice have intestinal flora disturbance and probiotics can participate with the occurrence and development of the disease by adjusting metabolin.Bifid triple viable capsule is currently one of the most widely used probiotics preparation clinically and includes long Bifidobacterium,lactobacillus acidophilus and enterococcus faecalis.It has been proved that the conditions of SLE mice can be improved by adjusting intestinal flora.But there are no related reports on whether Bifid triple viable capsule participates with the control of SLE by adjusting metabolism.ObjectiveThe research will discuss the differences of metabolome features of SLE patients,healthy people,SLE patients before and after probiotics-assisted treatment and before and after traditional treatment,and look for the plasma metabolin relating to the disease and probiotics treatment and the possible influenced metabolic pathway,in order to further reveal the mechanism of probiotics-assisted treatment participating with SLE adjustment and control and provide a theoretical basis for probiotics-assisted treatment.MethodsThe research has 18 objects of research divided into probiotics group(n=6),traditional treatment group(n=6)and healthy control group(n=6).Probiotics group was given hormone,Mycophenolate Mofetil,hydroxychloroquine and probiotics(Bifid triple viable capsule(0.63 g bid))treatment.Traditional treatment group was given hormone,Mycophenolate Mofetil and hydroxychloroquine treatment.The observation period is three months.The diagnosis of all SLE patients conforms to 2019 EULAR/ACR SLE classification standards(SLEDAI-2000≤6).HC group consists of the healthy people who received our physical examination and has the required age and gender.The baseline information(including gender,age,laboratory indexes and SLEDAI-2000)of objects of research when being recruited was collected.Meanwhile,the blood samples of HC group and SLE patients before and after probiotics treatment and traditional treatment were collected for non-targeted metabonomics analysis.Results1.The patients of probiotics group didn’t have serious adverse reactions before and after treatment in the whole experiment observation period,which is in line with traditional treatment group.2.The results of whole level analysis of non-targeted metabonomics show that 1066 chemical compounds were detected in the blood samples for test mainly including lipid and lipid-like molecules.PSL-DA analysis shows that SLE patients and healthy control group,before and after treatment of traditional treatment group,before and after treatment of probiotics group,and after probiotics treatment and after traditional treatment have a good sample grouping in terms of whole metabolome features,meaning the metabolins among groups have obvious differences.3.Difference metabolin analysis shows that with variable importance projection(VIP)>1,fold change(FC)>1.2 or<0.833 and P<0.05 as screening conditions,compared with HC group,probiotics group had 134 much different metabolins(68 up-regulated metabolins and 66 down-regulated metabolins)before treatment;probiotics group has 23 much different metabolins before and after treatment(5 up-regulated metabolins and 18 down-regulated metabolins).Similarly,before treatment,traditional treatment group and HC group have 94 much different metabolins(53 up-regulated metabolins and 41 down-regulated metabolins);there are 50 much different metabolins before and after traditional treatment(32 up-regulated metabolins and 18 down-regulated metabolins).The results show that the two treatment methods may influence the changes of plasma metabolins.Among them,probotics group before and after treatment and traditiona treatment group before and after treatment only have three common difference metabolins4.According to the strict self-defined screening standards,difference metabolins should conform to the following change trend,namely having more metablolins than HC group before treatment and having more metabolins than after treatment.And before treatment,in 68 obviously up-regulated metabolins compared with HC group,probiotics group,20-Carboxy-Leukotriene B4,phosphatidylcholine(14:0e/21:2)(PC(14:0e/21:2))and 12(S)-HETE and phosphatidyl ethanolamine(18:1e/22:4)(PE(18:1e/22:4))conform to the above screening standards;and ROC curve analysis shows that the AUC values of probiotics group before treatment and HC group are both higher than 0.7 proving its diagnosis and treatment efficacy.The difference metabolins of traditional treatment group before and after treatment don’t conform to the above screening standards.5.The enrichment analysis of difference metabolin KEGG shows that the metabolic pathways for the co-enrichment of the difference metabolins between probiotics group before treatment and HC group,and before and after probiotics treatment include ovary steroid generation,arachidonic acid metabolism and aldosterone synthesis and secretion.However,no pathway for the common enrichment of difference metabolins between before traditional treatment and HC group,and before and after traditional treatment was found.12(S)-HETE was found in arachidonic acid metabolism and aldosterone synthesis and secretion.And the synthesis of arachidonic acid is closely related to inflammatory response.Aldosterone has a potential immunoregulation function and may play a role in the onset and progress of lupus nephritis.The generation pathway of aldosterone synthesizes and secretes various steroid hormones including estrogen,progesterone,androgen and its predecessor.Conclusions1.SLE patients and healthy control group,SLE patients before and after probiotics treatment,and before and after traditional treatment have much different metabolins.And probiotics-assisted treatment may adjust SLE to some extent by adjusting the balance between 20-Carboxy Leukotriene B4,PC(14:0e/21:2),12(S)-HETE and PE(18:1e/22:4).And the four metabolins are also the potential biological biomarkers for SLE diagnosis.2.The research also found three metabolic pathways relating to probiotics-assisted treatment,which may provide new clues for exploring the mechanism of probiotics treating SLE. |
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