Ziwan-Taoren herb pair can exert an therapeutical effect in primary Sjogren’s syndrome through inhibiting the TLR/NF-κB pathway

Background:Ziwan and Taoren(ZT)is a classic medicine pair in the formula of Mai Dong Di Shao Decoction,has been used to treat primary Sjogren’s syndrome(pSS)for more than 20 years.But its action mechanism is still unknown.This study is aimed to reveal the potential mechanism of ZT treated pSS and discover its active compounds of ZT and therapeutic target for pSS.Methods:Firstly,the potential pathways of ZT for pSS treatment were predicted through network pharmacology and GO and KEGG enrichment analysis.Secondly,the inter-structural relationships between active compounds of ZT and target proteins were visualized using molecular docking techniques.Finally,efficacy and mechanism were conducted through in vivo experiments,such as water intake,spleen index,hematoxylin-eosin staining pathological changes,ELISA,Western Blot analysis,and immunofluorescence staining.Results:Nine active compounds were extracted from network pharmacology,including quercitrin,luteolin,kaempferol,β-sitosterol,isorhamnetin,galangin,hederagenin,diosmetin and gibberellin 7.Seven disease targets were identified:RELA,TP53,AKT1,interleukin(IL)6,MAPK1,ESR1,IL10;with RELA being the most core target.KEGG and GO enrichment analysis indicated that ZT may act through the TLR/NF-κB/RELA inflammatory mechanism process.preliminary results of molecular docking showed that ZT’s active compounds bind well to the RELA(p65)receptor.In vivo results demonstrated that a high dose of ZT significantly improved water intake and reduced lymphocytes infiltration in submandibular gland pathology in NOD mice.The expression content of AQP5 and vasoactive intestinal peptide in the submaxillary gland was significantly increased,while levels of inflammatory factors such as tumor necrosis factor-α,IL-6,and IL-1βalong with protein expressions including toll-like receptor4,p-p65 and p-IKKα/βin NF-κB pathway were reduced.Conclusions:The ZT treatment exhibits a promising efficacy in mitigating dryness symptoms of pSS,potentially attributed to its capacity for suppressing the TLR/NF-κB inflammatory signaling pathway.

Global Protein Expression Analysis of Molecular Markers of DS-1-47,a Component of Implantation-promoting Traditional Chinese Medicine

This study investigated the molecular markers of DS-1-47,a component of an implantation-promoting traditional Chinese medicine consisting of Astragalus mongholicus,Atractylodes macrocephala,Scutellaria baicalensis and Dipsacales,in an attempt to clarify the molecular mechanism and action targets of DS-1-47.Controlled ovarian stimulation（COS） method was used to establish the implantation dysfunction models of mice.Animals were divided into normal pregnant group,COS model group and DS-1-47 group.Laser capture microdissection-double dimensional electrophoresis-mass spectrum（LCM-DE-MS） was used to analyze the uterine protein molecules that were possibly involved in the promotion of implantation.Twenty-three proteins in DS-1-47 group were significantly changed as compared to those in COS model group,with 7 proteins down-regulated and 16 proteins up-regulated.Except for some constituent proteins,the down-regulated proteins included collagen α-1（Ⅵ） chain,keratin 7,keratin 14,myosin regulatory light chain 12 B,myosin light polypeptide 9,heat shock protein β-7,and C-U-editing enzyme APOBEC-2;the up-regulated proteins included apolipoprotein A-I,calcium regulated protein-3,proliferating cell nuclear antigen,L-xylulose reductase,and calcium binding protein.These 23 proteins that were regulated by DS-1-47 represented a broad diversity of molecule functions.The down-regulated proteins were associated with stress and immune response,and those up-regulated proteins were related to proliferation.It was suggested that these proteins were important in regulating the uterine environment for the blastocyst implantation.By identification of DS-1-47 markers,proteomic analysis coupled with functional assays is demonstrated to be a promising approach to better understand the molecular mechanism of traditional Chinese medicine.