HuoguⅠformula prevents steroid-induced osteonecrosis in rats by down-regulating PPARγ expression and activating Wnt/LRP5/β-catenin signaling

OBJECTIVE: To investigate the effects of HuoguⅠformula on regulation of lipid metabolism in steroid-induced osteonecrosis of the femoral head(SONFH) rats and verify our hypothesis that HuoguⅠ formula regulates lipid metabolism by down-regulating peroxisome proliferator-activated receptor gamma(PPARγ) expression and activating Wnt signaling pathways.METHODS: Eighty-five rats were divided into four groups: control, model, Huogu 15 g/kg and Huogu30 g/kg. Six weeks later, animals were anaesthetized, femora were dissected for histopathological examination of the osteonecrotic changes and repair processes, micro computed tomography(Micro-CT)-based micro-angiography was performed to assess vascularization. Serum lipid levels were detected by haematological examination. The expressions of PPARγ, Wnt3a, low density lipoprotein receptor-related protein 5(LRP5) and β-catenin were evaluated by immunohistochemistry,Western blot and quantitative real-time polymerase chain reaction analyses.RESULTS: The incidence of osteonecrosis, ratio of empty lacuna, adipose tissue area and adipocyte perimeter in the bone marrow were dramatically lower in the Huogu Ⅰ formula treatment groups.Bymicro-CTquantification,HuoguⅠformulatreatment dose-dependently increased vessel volume,vessel surface, percentage of vessel volume and vessel thickness of the femoral heads of SONFH rats. Levels of serum lipid in Huogu 15 g/kg and Huogu 30 g/kg groups reduced significantly.HuoguⅠformula treatment could suppress the expression of PPARγ and increase the expressions of Wnt3a, LRP5 and β-catenin at both protein and mRNA levels.CONCLUSION: The results of our present study highlight the lipid-lowering potential of Huogu Ⅰformula, and provide further evidence of the involvement of the PPARγ inhibition and Wnt/LRP5/β-catenin signaling activation in the effects of HuoguⅠformula.

OBJECTIVE： To investigate the effects of Huogu I formula on regulation of lipid metabolism in ste- roid-induced osteonecrosis of the femoral head （SONFH） rats and verify our hypothesis that Huogu I formula regulates lipid metabolism by down-regulating peroxisome proliferator-activated receptor gamma （PPARy） expression and activating Wnt signaling pathways. METHODS： Eighty-five rats were divided into four groups： control, model, Huogu 15 g/kg and Huogu 30 g/kg. Six weeks later, animals were anaesthe- tized, femora were dissected for histopathologicalexamination of the osteonecrotic changes and re- pair processes, micro computed tomography （Mi- cro-CT）-based micro-angiography was performed to assess vascularization. Serum lipid levels were detected by haematological examination. The ex- pressions of PPARy, Wnt3a, low density lipoprotein receptor-related protein 5 （LRP5） and 13-catenin were evaluated by immunohistochemistry, Western blot and quantitative real-time polymerase chain reaction analyses. RESULTS： The incidence of osteonecrosis, ratio of empty lacuna, adipose tissue area and adipocyte perimeter in the bone marrow were dramatically lower in the Huogu ~ formula treatment groups. By micro-CT quantification, Huogu ~ formula treat- ment dose-dependently increased vessel volume, vessel surface, percentage of vessel volume and vessel thickness of the femoral heads of SONFH rats. Levels of serum lipid in Huogu 15 g/kg and Huogu 30 g/kg groups reduced significantly. HuoguⅠformula treatment could suppress the ex- pression of PPARy and increase the expressions of Wnt3a, LRP5 and 13-catenin at both protein and mRNA levels. CONCLUSION： The results of our present study highlight the lipid-lowering potential of Huogu Ⅰ formula, and provide further evidence of the in- volvement of the PPARy inhibition and Wnt/LRPS/ 13-catenin signaling activation in the effects of Huogu Ⅰ formula.