Analysis Of High Risk Indicators Of GA-ONFH And Study On Yougui Yin Regulating Exosomes From BMSC In The Treatment Of GA-ONFH

Objective:Based on the serum metabolites,the potential risk indicators of GA-ONFH were screened by binary logistic regression analysis,which can help the patients who use hormones to find the disease in time and make a clear diagnosis,so as to take the best treatment measures.By matching the high-risk indicators obtained from the study with the basic research results of Youguiyin intervening in the expression of exosomes derived from BMSC,the potential mechanism of Youguiyin intervening in early GA-ONFH patients is explored through bioinformatics database,which provides modern medical basis for Youguiyin treating GA-ONFH according to the kidney tonifying theory of traditional Chinese medicine.Methods:1.Clinical study:according to the inclusion and exclusion criteria,60 patients admitted to the Department of orthopedics and traumatology of Jiangsu Hospital of traditional Chinese Medicinefrom September 2018 to September 2020 were selected as the observation group,and 60 patients without femoral head necrosis in the rheumatology department of Jiangsu Hospital of traditional Chinese Medicine during the same period(September 2018 to September 2020)were selected as the observation group Control group.The basic information,blood routine,liver and kidney function were recorded.Spss25.0 statistical software was used for binary logistic regression analysis of the above information,and GA-ONFH and other high-risk indicators were screened out.2.Experimental study:12 rats were randomly divided into Youguiyin group,blank control group and model group,with 4 rats in each group.Youguiyin group and model group used horse serum combined with dexamethasone injection to make early femoral head necrosis model.Four weeks after modeling(from the fifth week),Youguiyin group was given Youguiyin Decoction by gavage,20ml/kg daily.Hormone control group and blank control group were given normal saline by gavage,20ml/kg daily.The three groups were intervened for 6 weeks.Then the rats were killed under excessive anesthesia,and the bone marrow mesenchymal stem cells were cultured.Then 20 ml of cell supernatant was taken,and exosomes were extracted by centrifugation.Then the exosomes were identified by transmission electron microscopy.Western blot was used to detect the expression of exosomes and exosomes specific protein CD63.Finally,immunofluorescence was used to verify the expression of exosomes specific protein.Finally,the above results were matched with some positive results of clinical research by bioinformatics,and analyzed by KEGG and go.Results:1.Clinical research:independent sample t-test analysis showed that:white blood cell count(P=0.228),ALP(P=0.005),Afu(P=0.000),glutamate dehydrogenase(P=0.038),urea(P=0.000),sugar(P=0.030),serum bicarbonate(P=0.033),Mg(P=0.001),lipase(P=0.003)had statistical significance,the rest had no statistical significance.After binary logistic regression analysis,ALP(P=0.005,or=1.031)and urea(P=0.005,or=1.740)were the risk indicators of femoral head necrosis.2.Experimental study:compared with the model group,the value of BMSCs and BMSCs exo in Youguiyin group increased significantly,and the content of CD63 in the model group was lower than that in the blank group,while the exosomes in Youguiyin group were significantly up-regulated.The results were consistent with WB.After KEGG analysis and go analysis,the potential targets and pathways of Youguiyin regulating exosomes to intervene GA-ONFH were obtainedConclusions:ALP and urea are high risk indicators of steroid induced osteonecrosis of the femoral head.Youguiyin can potentially interfere with the osteogenic activity of BMSC by regulating the secretion of exosomes,and then play its therapeutic role.This effect may be related to the regulation of cell adhesion molecule binding,integrin binding and cadherin binding pathway.