Study On The Protective Mechanism Of Zuogui Pill For Secondary Osteoporosis After Endocrine Treatment Of Breast Cancer

Objective : To study the protective mechanism of Zuogui Pill on secondary osteoporosis after endocrine therapy of breast cancer.Methods : Modeling: a mouse model of secondary osteoporosis after endocrine therapy for breast cancer was established by referring to the literature.Ovariectomize(OVX)was performed in all the other groups except the pseudo-castrating group.After 10 weeks,groups of mice were implanted with murine4T1 cell line to establish a mouse model of breast cancer.Tumors larger than 0.3cm were removed after 10 days,and tumor tissues were collected for pathological analysis.Mice without tumor recurrence were selected and divided into groups.Grouping: 84 SPF female breast cancer in BALB/c mice were randomly divided into 7 groups,false castration group(Sham),blank(Ovariectomize,OVX),model group(OVX,letrozole),Alendronate sodium group(OVX,letrozole,ALN),left to high dose group of zuogui pills(OVX,letrozole,zuogui pills-H),left to bolus dose group of zuogui pills(OVX,letrozole,zuogui pills-M),left to pill low dose group zuogui pills(OVX,letrozole,zuogui pills-L),in addition to the false castration group,the blank group was given distilled water,The other groups were given the corresponding regimen for 4 weeks.Sampling and detection: After 4 weeks,blood was collected from the eyeball,and serum estradiol E2,bone alkaline phosphatase(BALP)and procollagen I amino terminal peptide(PINP)were detected by ELISA.After the execution,the right femur and tibia were taken for microtomography(CT)to detect in vitro bone density and trabecular microstructure,and HE staining for bone histomorphological observation.The left femur and tibia were transferred to liquid nitrogen bottles for quick-freezing,and then stored in the refrigerator at-80?.The expressions of proteins Wnt3 a and Runx2 were detected by Western blot.Results:1.Test results of serum E2 level and bone metabolism indexes BALP and PINP in miceCompared with the blank group,the serum E2 content in the castration group and the model group was significantly reduced,with a significant difference(P<0.05),and the model group was lower than the castration group(P<0.05);compared with the model group,Zuoguiwan There was no significant difference in serum E2 content between high,middle and low dose groups and alendronate sodium group(P>0.05).Compared with the blank group,the BALP and PINP contents in the castration group and model group were significantly increased,with significant differences(P<0.05).And the model group was higher than the castration group(P<0.05),indicating that the use of letrozole to further increase bone metabolism indexes after castration;compared with the model group,Zuoguiwan high,medium and low dose groups,alendronic acid The contents of BALP and PINP in the sodium group were significantly reduced(P<0.05).2.Observation results of mouse bone histomorphologyObservation under electron microscope: the bone tissue of the blank group mice showed that the trabecular bone structure was intact,the trabecular bone was connected evenly,and the morphology of the fat cells and bone cells was clear;The bone trabeculae of the castration group and the model group were sparsely and slenderly distributed,and the structure was incomplete,but the model group was more severe.The trabecula had poor continuity,obvious fractures,and increased fat cells.Increased diameter,indicating the effect of letrozole on bone quality after castration.Compared with the model group,the bone morphology of Zuogui Pill's high,medium and low dose group and alendronate group was not obvious.The trabecular bone structure was not continuous and the trabecular bone was connected evenly,Indicating that the intervention drugs all improve the changes of osteoporosis-like bone.3.Results of femur bone density and bone microstructure in miceCompared with the blank group,the bone volume fraction(BV/TV),trabecular bone thickness(Tb.Th),trabecular bone number(Tb.N),and bone mineral density(BMD)of the castration group and model group were significantly reduced(P<0.05),with statistical significance,and the trabecular bone separation(Tb.Sp)was significantly increased(P<0.05);compared with the model group,Zuogui Pill high,middle dose group,alendronate sodium The bone mineral density and bone microstructure of the group all improved,and the difference was statistically significant(P<0.05).There was no significant difference in the low dose of Zuoguiwan(P>0.05).4.Expression of Wnt3 a and Runx2 proteins in bone tissueCompared with the blank group,the Wnt3 a and Runx2 protein content of the femur in the castration group and the model group decreased significantly,which was statistically significant(P<0.05);compared with the model group,the Zuogui pill high,medium,and low dose groups,alen The content of Wnt3 a and Runx2 protein of femur in the sodium phosphonate group increased significantly(P<0.05).Conclusions:1.Zuogui Pill can reduce the content of BALP and PINP in serum of mice with secondary osteoporosis after endocrine therapy,promote bone formation,delay bone loss,and prevent osteoporosis.2.Zuogui pill can improve the bone microstructure of mice with secondary osteoporosis after endocrine therapy.3.Zuogui pill can up-regulate Wnt3 a and Runx2 proteins in mice with secondary osteoporosis after endocrine treatment,activate and regulate the protective mechanism of Wnt/?-catenin pathway,and promote osteogenic differentiation and bone formation.4.The high-dose,medium-dose and low-dose groups of zuogui pill interfered with mice with secondary osteoporosis after endocrine therapy for breast cancer respectively,and found that the high-dose group had the best effect.5.Zuogui pill has therapeutic effect on secondary osteoporosis after endocrine therapy for breast cancer.