The Therapeutic Mechanism Of Yunpi Jiedu Tongluo Qushi Recipe In Regulating Lipid Metabolism And In The Treatment Of Rheumatoid Arthritis

Objective Multi-dimensional mass spectrometry-based shotgun lipidomics(MDMS-SL)technique was used to qualitatively and quantitatively analyze the plasma lipids of rheumatoid arthritis(RA)patients and collagen-induced arthritis(CIA)rats,starting from the relationship between abnormal metabolism and RA disease progression.The intervention effect of Yunpi Jiedu Tongluo Qushi Recipe on the metabolism-related mechanism and the therapeutic mechanism of treating RA were explored.Methods 1.Pilot study on the characteristics and abnormal mechanism of plasma lipid metabolism in RA.30 RA patients and 30 healthy controls were recruited according to the 2010 ACR/EULAR classification standard.The basic information and clinical indexes of the subjects were recorded.The contents of serum TNF-α,TGF-β1 and IL-17A were detected by ELISA.The ratio of Th17/Treg in PBMC was detected by flow cytometry.And the expression of PI3K,AKT,P-AKT,STAT3 and P-STAT3 in peripheral blood PBMC was detected by Western Blot.The contents of TAG,SM,Car,HNE,PC,LPC and PI in plasma were measured by MDMS-SL,and the correlation between lipid contents and CRP or ESR contents was analyzed.2.Study on the therapeutic mechanism of Yunpi jiedu Tongluo Qushi Recipe in the treatment of-RA.32 Wistar rats were randomly divided into control group,model group,Chinese medicine group and western medicine group.CIA method was used to model rats in model group,Chinese medicine group and western medicine group.Chinese medicine group were given Yunpi jiedu Tongluo Qushi Recipe once a day,and western medicine group were given methotrexate(MTX)once a week.The same volume of distilled water was given to the control group and the model group.The rats were killed 5 weeks after the first immunization.During this period,the body weight and toe volume of both hindlimbs were measured once a week.The contents of serum TNF-α,TGF-β1 and IL-17A were detected by ELISA method.The contents of PI,PS,Cer,LPC,SM,PG and HNE in liver were detected by MDMS-SL.The correlation between liver lipid contents and serum TNF-α,TGF-β1 or IL-17A contents was analyzed.Results 1.The levels of CRP and ESR in RA group were beyond the normal range.The levels of TNF-α,TGF-β1 and IL-17A were not significantly different from those in HC group.And the ratio of Th17/Treg was increased in RA group.Compared with HC group,the contents of Car,HNE and LPC in plasma in RA group decreased significantly,while the content of PI increased significantly.There was no significant difference in the contents of TAG,SM and PC.There was a negative correlation between the content of LPC and the levels of CRP and ESR.In RA group,the expression of PI3K protein in PBMC decreased significantly,while the phosphorylation level of AKT and STAT3 had no significant change.2.Compared with the control group,the toes of both hind limbs of rats in the model group showed obvious swelling,and the toe swelling was relieved after treatment with Yunpi Jiedu Tongluo Qushi Recipe or MTX.There was no significant difference in ALT,AST and TP among the four groups,but Crea decreased in Chinese medicine group and western medicine group compared with the model group.At 21 or 35 days after the first immunization,the contents of serum TNF-α,TGF-β1 and IL-17A in the model group were significantly higher than those in the control group,and the contents of these cytokines decreased significantly after treatment with Chinese medicine.Compared with the control group,the contents of PI and PS lipids in the model group increased,while the contents of Cer,LPC,SM,PG and HNE lipids had no significant change.After treatment with Chinese medicine or western medicine,the content of PI decreased significantly,the content of PS did not change significantly,and the contents of LPC,HNE,SM and Cer increased significantly.There was a significant positively correlation between the content of PI lipids and the contents of TNF-α,TGF-β1 and IL-17A in serum of rats.Conclusions 1.The abnormal plasma lipid metabolism in RA patients was manifested as the decrease of Car,HNE and LPC,the increase of PI content and the increase of PC/LPC ratio,indicating that the abnormal lipid metabolism in RA patients was related to energy metabolism,inflammatory response and cellular metabolism.2.The results of animal experiments were similar to those of clinical samples,and the content of PI lipids in the liver of CIA rats was significantly increased compared with the control group,and was significantly positively correlated with serum inflammatory factors,indicating that the abnormal increase of PI content in RA can affect the PI3K-mediated inflammatory pathway,and then affect the condition of RA.3.After the treatment of Yunpi Jiedu Tongluo Qushi Recipe,the joint swelling of CIA rats was relieved,the inflammatory factors decreased,and the content of PI lipids in liver decreased,indicating that Yunpi Jiedu Tongluo Qushi Recipe may regulate the release of inflammatory factors,and alleviate the disease by regulating the metabolism of PI lipid molecules and affecting PI3K-related signal pathways.