Therapeutic Effect And Mechanism Of Chinese Herbal Compound Shenyi On Lupus Nephritis In MRL/lpr Mice

Background and Objective: Lupus nephritis(LN)is one of the most serious and common complications of systemic lupus erythematosus(SLE),and its clinical symptoms are mainly immune hyperactivity and renal function damage.Most lupus nephritis patients eventually progress to endstage renal disease.At present,there is no effective drug for the treatment of lupus nephritis.The clinical treatment is mainly hormone pulse therapy and immunosuppressive therapy.However,the use of the two main drugs is accompanied by a variety of serious side effects,and It can aggravate kidney damage,and it has research significance to find low-side effects and high-efficiency drugs for lupus nephritis.Studies have shown that the traditional Chinese medicine compound Shenyi corrects the imbalance of the proportion of T cells in the body,regulates immunity,and inhibits glomerulosclerosis.The pathogenesis of LN is mainly immune hyperactivity,accompanied by the risk of end-stage renal disease progressing to glomerulosclerosis.SY regulates the body’s immunity and delays glomerular sclerosis whether it has practical significance.The purpose of this study was to establish a classic animal model of LN-MRL/lpr mice,to study the therapeutic effect of Compound Shenyi on lupus nephritis in MRL/lpr mice and to explore its therapeutic mechanism.Methods: Study 1: Compound Shenyi improves the symptoms and signs of lupus nephritis in MRL/lpr mice: 35 MRL/lpr mice were randomly divided into a model group(MRL/lpr group,n=8)and a low-dose Shenyi group(low SY dose group,n=9),Shenyi middle dose group(SY medium dose group,n=9),and Shenyi high dose group(SY high dose group,n=9).C57 mice were the normal control group(n=6).The SY groups were given15.34g/kg/day,46.02g/kg/day,92.04g/kg/day by gavage,and the normal control group and MRL/lpr group were given 0.5ml/day normal saline by gavage.Intervention started at 12 weeks of age,once a day for 14 weeks.The animal signs,renal pathology,urine protein-to-creatinine ratio(u PCR),serum antinuclear antibody(ANA),anti-double-stranded DNA antibody(anti-ds DNA)concentration,renal Ig G,Ig G1,Ig M,C3 immune deposition fibrin expression in kidney tissue were observed in each group;Study 2:Compound Shenyi exerts its therapeutic effect on lupus nephritis by inhibiting complement activation in MRL/lpr mice: 35 MRL/lpr mice were randomly divided into model group(MRL/lpr group,n=8),Shenyi low Dose group(SY low-dose group,n=9),Shenyi middle-dose group(SY medium-dose group,n=9),and Shenyi high-dose group(SY high-dose group,n=9).C57 mice were the normal control group(n=6).The SY groups were given 15.34g/kg/day,46.02g/kg/day,92.04g/kg/day by gavage,and the normal control group and MRL/lpr group were given0.5ml/day normal saline by gavage.Intervention started at 12 weeks of age,once a day for 14 weeks.The immune deposition of C4 and C5b-9 in the kidneys of animals in each group and the expression of complement factor proteins in the kidneys were observed;Study 3: The complement factor B inhibitor LNP023 improves lupus nephritis in MRL/lpr mice: MRL/lpr mice were randomly divided into MRL/lpr model group(n=6)and LNP023group(n=6),C57 mice were Normal control group(n=6).The LNP023 group was intragastrically administered with LNP023 solution(30mg/kg/d)for 2 weeks;the MRL/lpr model group was intragastrically administered with Na Cl solution(0.5ml/d)for 2 weeks;the normal control group was intragastrically administered with Na Cl solution(0.5ml/d)for 2weeks.The external signs,renal pathology,renal function,renal immune complex and complement deposition,serum anti-ds DNA,serum ANA concentration and the expression of the core complement factors of the alternative complement pathway were analyzed in the three groups;Study4: Exploring the molecular mechanism of traditional Chinese medicine compound Shenyi in the treatment of lupus nephritis based on network pharmacology: using TCMSP database and Shanghai Institute of Organic Medicine and chemical composition database to screen the active ingredients in SY,using TCMSP database and Swiss Target Prediction database to collect SY active ingredient targets;Use the Gene Cards database and the OMIM database to screen LN disease targets,map the intersection of component targets and disease targets,and obtain therapeutic targets.GO and KEGG enrichment analysis of therapeutic targets was performed using the METASCAPE database.Results:(1)Compared with the MRL/lpr model group,the lupus-like signs of animals in each dose group of SY intervention were reduced,and the renal pathology showed reduced immune complex deposition,inflammatory cell infiltration,mesangial cell proliferation,and u PCR,serum ANA,anti-ds DNA concentration decreases.,And the deposition of Ig G,Ig G1,Ig M,and C3 in renal tissue decreased;(2)Compared with the MRL/lpr model group,the immune deposition of C4 and C5b-9 in the kidney tissue of animals in each dose group of SY intervention was reduced.The expressions of renal complement factors C3,C5 and CD35 decreased,while the expressions of C5 a and C5 b increased;(3)Compared with the MRL/lpr group,the lupus-like signs in the LNP023 group were alleviated,the renal function was improved,the serum anti-ds DNA and ANA concentrations were decreased,and the renal deposition of Ig M,Ig G,Ig G1,C1 q,C3,and C4 was alleviated.Renal pathology showed that LNP023 could alleviate the renal pathological damage in MRL/lpr mice.Compared with the MRL/lpr model group,the treatment group had no crescent formation,less immune deposition,no nuclear fragmentation,and less inflammatory cell infiltration.The expression of complement proteins C3,C3 b,CR1,CFB and C5b-9 in kidney tissue and liver decreased,and the expression of C5 increased;(4)There are 77 active ingredients in SY,497 active ingredients,1112 LN disease targets,and 236 SY treatment LN core targets.Among them,17 core targets,including AKT1,TNF,IL6,and TP53.GO and KEGG enrichment analysis showed that SY may play a role in the treatment of LN through "AGE-RAGE","PI3K-Akt","IL-17" and other pathways.Conclusion: The traditional Chinese medicine compound Shenyi improves the symptoms and signs of lupus nephritis in MRL/lpr mice.Its mechanism of action is to inhibit the activation of the complement pathway.The complement factor B inhibitor LNP023 inhibits the activation of the alternative complement pathway and improves lupus nephritis in MRL/lpr mice.SY may also play a role in the treatment of LN by regulating "AGERAGE","PI3K-Akt","IL-17" and other signaling pathways.