Study On The Mechanismof Bone Marrow Adipogenesis-mediated Lipotoxicity In Osteoporosis And The Interventional Effect Of Drynaria Rhizome And Chinese Patent Medicine Containing Drynaria Rhizome

PART 1 The mechanism of Drynaria Rhizome on reducing bone marrow fat in ovariectomy-induced bone loss ratsObjective:To observe the effect of Drynaria Rhizome on improving bone marrow fat in ovariectomy-induced bone loss ratsMethods:Eighteen SD rats were randomly divided into sham operation group(SHAM),ovariectomized group(OVX),and Drynaria Rhizome group(OVX-DR)group.After the model was established,each group was given corresponding administration for 3 months,Then the organ weight and fat metabolism indexes of rats in each group were detected.The distribution of femur trabecular bone and the content of adipose tissue in bone marrow cavity were observed by section staining.The expression of ?-Catenin and PPARy protein in femur was detected by immunohistochemistry.Results:Compared with ovariectomized group,Drynaria Rhizome group can effectively improve the blood lipid content of rats,but it does not have the effect of inhibiting uterine atrophy.It significantly increased the number of femur trabeculae and and decreased the fat content of bone marrow,And it increases the expression of ?-Catenin protein of femur,and reverses the high expression of PPARy in ovariectomized bone tissue to a certain extent.Conclusion:Drynaria Rhizome may inhibit bone marrow adipogenesis and improve ovoectomy-induced bone loss by regulating the balance of ?-Catenin-PPARy signal crosslinking axis.PART 2 The clinical effect of Chinese patent medicine containing Drynaria Rhizome in the treatment of primary osteoporosis and the effect of preventing refractureObjective:To observe the clinical efficacy of Gubao capsule in the treatment of primary osteoporosis and the effect of preventing re-fracture.Methods:Patients with osteoporosis who were treated in the Department of Orthopedics,Affiliated Hospital of Shandong University of Traditional Chinese Medicine from June 2018 to February 2020 were divided into two groups.The control group was treated with conventional methods,and the observation group was additionally treated with Gubao capsule.The patients were followed up for 1 year.During the period,the treatment effect and medication safety of the two groups were comprehensively evaluated by comparing the bone mineral density,clinical symptom score,the incidence of refracture and the improvement degree of osteoporosis between the two groups.Results:There was no significant difference in baseline data between the two groups.Conventional anti-osteoporosis therapy or the addition of Gubao capsule could effectively delay bone loss.Compared with the control group,the observation group significantly improved the TCM syndromes and clinical symptom scores of the patients,the VAS and ODI scores were lower,and the incidence of re-fracture was also lower.In addition,the results of this study showed that no obvious adverse reactions occurred when taking Gubao capsule.Conclusion:The addition of Gubao capsule to conventional anti-osteoporosis treatment can effectively improve the clinical symptoms of patients,improve the quality of life,reduce the incidence of re-fractures,and the application is safe and reliable.PART 3 The mechanism of Gubao capsule in reducing lipotoxicity and promoting osteogenic differentiationObjective:To study the mechanism of Gubao capsule in reducing lipotoxicity and promoting osteogenic differentiation.Methods:Screen the safe concentration of Gubao capsule on MC3T3-E1 and 3T3-L1 cells,and then intervene MC3T3-E1 cells induced by osteogenesis.The expression of ALP in the cell culture supernatant was detected at 3/5/7 days,and Alizarin red staining was performed at 18 days to analyze the effect of Gubao capsule extract on osteogenic differentiation.Then select the optimal concentration that promotes osteogenic differentiation to intervene in MC3T3-E1 cells,and extract the total cell protein at 10d for the detection of osteogenic and Wnt/?-cateninpathway-related proteins.Then established a lipogenic/osteogenic co-culture system,and used Gubao capsule to interfere the lower layer of osteoblasts in the co-culture system,and the ALP content in the cell culture supernatant was detected at 5 days,the alizarin red staining was performed at 14 days,and the total protein of the cells was extracted for detection 10 days after the intervention.Results:Low concentration of Gubao capsule extract can effectively promote the expression of ALP in MC3T3-E1 cells,increase the content of calcium nodules,and corresponding significantly increase the expression of osteogenic proteins such as Runx2,Osterix and OPN in MC3T3-E1 cells,and promote the expression of Wnt3a and ?-catenin proteins in Wnt/?-catenin pathway.Compared with the blank group,the osteogenic differentiation activity of MC3T3-E1 in the co-culture group was significantly decreased,and the expression of ?-catenin protein was decreased,and the expression of PPAR?protein was promoted.After the treatment with Gubao capsule,the osteogenic differentiation ability of MC3T3-E1 and the expression of ?-catenin protein were significantly improved compared with the co-culture group,and the expression of PPAR?protein was decreased accordingly.Conclusion:This study found that the low-dose Gubao capsule extract may promote the expression of osteoblast-related proteins by activating the Wnt/?-catenin pathway,and then promote osteoblastic differentiation and maturation.It may protect or restore the damage of osteoblasts caused by lipotoxicity by regulating the signaling axis of ?-catenin-PPARy,and promote osteoblastic differentiation.