Objective To observe the effect of Wenshen Tongluo Zhitong Recipe(WTZR)on adipocyte exosomes on bonefat balance in osteoporosis model mice.Methods Totally 80 Balb/c mice were randomly divided into sham operation group...Objective To observe the effect of Wenshen Tongluo Zhitong Recipe(WTZR)on adipocyte exosomes on bonefat balance in osteoporosis model mice.Methods Totally 80 Balb/c mice were randomly divided into sham operation group(Sham),model group(Ovx),exosome group(EXO),and WTZR intervention exosome group(EXO+WTZR).An ovariectomized osteoporosis mouse model was constructed.The Sham and OVX group were injected physiological saline into the caudal vein at a dose of 200μL respectively,and EXO and EXO+WTZR group were injected exosomes into the caudal vein at a dose of 30μg(200μL),once a week for 12 weeks.The contents of bone tissue-specific alkaline phosphatase(B-ALP),osteocalcin(OCN),tartrate-resistant acid phosphatase(TRAP),and interleukin-6(IL-6)in serum were detected by ELISA,and the levels of triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),and high density lipoprotein cholesterol(HDL-C)in serum were detected by a fully automatic biochemical analyzer.The histopathological changes of femur were observed by HE staining and TRAP staining under optical microscope,and the number of osteoclasts(N.Oc/BS)and the ratio of osteoclasts surface area to bone surface(Oc.S/BS)were calculated.Micro-CT was used to detect the microstructure of mouse femur bone[trabecular separation(Tb.Sp),bone mineral density(BMD),bone volume fraction(BV/TV)J and the distribution of surrounding adipose tissue[regulated bone marrow adipose tissue(rMAT and constitutive bone marrow adipose tissue(cMAT)J,and the three-dimensional reconstruction image analysis was performed.IHC and qPCR methods were used to detect the expression of mouse Runt-related transcription factor 2(RUNX2),osteogenesis-related transcription factor(Osterix),CCAT enhancer binding proteinα(CEBP-α)and peroxisome proliferation-activated receptor 2(PPAR 2)protein and mRNA in bone tissue,respectively.Results Compared with the Sham group,the levels of BALP,OCN,TRAP,IL-6,TC,TG,LDL-C,rMAT,and cMAT increased,the mRNA and protein levels of RUNX2 and Osterix increased(P<0.05,P<0.01),the content of HDL-C decreased(P<0.05),and the expressions of CEBP-α,PPAR γ2 mRNA and protein decreased(P<0.01)in the OvX group.Moreover,in the OvX group,trabecular bone was destroyed,the reticular structure was broken,the number of trabecular bone was reduced,the bone was lost,the bone microstructure was destroyed,and the Tb.Sp,BMD,BV/TV decreased(P<0.01).Compared with OVX group,the levels of B-ALP,OCN,and HDL-C in serum increased,the mRNA and protein of Tb.Sp,BMD,BV/TV,CEBP-α,PPAR 2 increased(P<0.05,P<O.O1),and the content of CHOL,TG,LDL-C,the proportion of rMAT,cMAT,and the expressions of RUNX2,Osterix mRNA and protein decreased(P<0.05,P<0.01)in the EXO and EX0+WTZR group.Moreover,in the EXO and EXO+WTZR groups,the number of trabeculae increased,the gap narrowed,and the fracture situation improved.Conclusion Adipocytederived exosomes can regulate bone-fat metabolism in osteoporotic,mice,improve bone microstructure,and reduce the formation of bone marrow adipose tissue.In addition,WTZR may exert an anti-osteoporotic effect by regulating bone-fat balance.展开更多
文摘Objective To observe the effect of Wenshen Tongluo Zhitong Recipe(WTZR)on adipocyte exosomes on bonefat balance in osteoporosis model mice.Methods Totally 80 Balb/c mice were randomly divided into sham operation group(Sham),model group(Ovx),exosome group(EXO),and WTZR intervention exosome group(EXO+WTZR).An ovariectomized osteoporosis mouse model was constructed.The Sham and OVX group were injected physiological saline into the caudal vein at a dose of 200μL respectively,and EXO and EXO+WTZR group were injected exosomes into the caudal vein at a dose of 30μg(200μL),once a week for 12 weeks.The contents of bone tissue-specific alkaline phosphatase(B-ALP),osteocalcin(OCN),tartrate-resistant acid phosphatase(TRAP),and interleukin-6(IL-6)in serum were detected by ELISA,and the levels of triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),and high density lipoprotein cholesterol(HDL-C)in serum were detected by a fully automatic biochemical analyzer.The histopathological changes of femur were observed by HE staining and TRAP staining under optical microscope,and the number of osteoclasts(N.Oc/BS)and the ratio of osteoclasts surface area to bone surface(Oc.S/BS)were calculated.Micro-CT was used to detect the microstructure of mouse femur bone[trabecular separation(Tb.Sp),bone mineral density(BMD),bone volume fraction(BV/TV)J and the distribution of surrounding adipose tissue[regulated bone marrow adipose tissue(rMAT and constitutive bone marrow adipose tissue(cMAT)J,and the three-dimensional reconstruction image analysis was performed.IHC and qPCR methods were used to detect the expression of mouse Runt-related transcription factor 2(RUNX2),osteogenesis-related transcription factor(Osterix),CCAT enhancer binding proteinα(CEBP-α)and peroxisome proliferation-activated receptor 2(PPAR 2)protein and mRNA in bone tissue,respectively.Results Compared with the Sham group,the levels of BALP,OCN,TRAP,IL-6,TC,TG,LDL-C,rMAT,and cMAT increased,the mRNA and protein levels of RUNX2 and Osterix increased(P<0.05,P<0.01),the content of HDL-C decreased(P<0.05),and the expressions of CEBP-α,PPAR γ2 mRNA and protein decreased(P<0.01)in the OvX group.Moreover,in the OvX group,trabecular bone was destroyed,the reticular structure was broken,the number of trabecular bone was reduced,the bone was lost,the bone microstructure was destroyed,and the Tb.Sp,BMD,BV/TV decreased(P<0.01).Compared with OVX group,the levels of B-ALP,OCN,and HDL-C in serum increased,the mRNA and protein of Tb.Sp,BMD,BV/TV,CEBP-α,PPAR 2 increased(P<0.05,P<O.O1),and the content of CHOL,TG,LDL-C,the proportion of rMAT,cMAT,and the expressions of RUNX2,Osterix mRNA and protein decreased(P<0.05,P<0.01)in the EXO and EX0+WTZR group.Moreover,in the EXO and EXO+WTZR groups,the number of trabeculae increased,the gap narrowed,and the fracture situation improved.Conclusion Adipocytederived exosomes can regulate bone-fat metabolism in osteoporotic,mice,improve bone microstructure,and reduce the formation of bone marrow adipose tissue.In addition,WTZR may exert an anti-osteoporotic effect by regulating bone-fat balance.
