Objective To observe the effects of Zhuang Gu Zhi Tong Formula (ZGZTF) on antagonist SOST in canonical Wnt/β-catenin signaling pathway in osteoporosis. Methods We analyzed the differential genes of patients with oste...Objective To observe the effects of Zhuang Gu Zhi Tong Formula (ZGZTF) on antagonist SOST in canonical Wnt/β-catenin signaling pathway in osteoporosis. Methods We analyzed the differential genes of patients with osteoporosis and normal subjects from the GEO database and then found SOST with specific expression. We analyzed SOST as an antagonist of the Wnt signaling pathway by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Then we studied the effect of ZGZTF on SOST in Wnt signaling pathway. Osteoporosis model was induced by ovariectomy (OVX) in 8-week-old female Sprague–Dawley (SD) rats. After 12 weeks of treatment with ZGZTF by intragastric administration, the rats were put to death in batch. The changes of alkaline phosphatase (ALP), bone gla protein (BGP) and estradiol (E2) in serum were determined, and bone mineral density (BMD) and histomorphology of right femur were observed. Biomechanics of lumbar vertebra were measured, and the expression of SOST, Wnt3a,β-catenin, LRP5, Runx2, Osx and their mRNA involving the canonical Wnt/β-catenin signaling pathway were detected by Western blot, RT-PCR and Immunohistochemical analysis. All data were analyzed by SPSS 22.0. Results Twelve weeks of treatment with ZGZTF could significantly decrease the level of ALP and BGP in serum, increase the BMD of femurs, and improve the biomechanical capability of vertebral body in maximum loading and elastic modulus. Concerning histomorphology, we found ordered arrangement of trabeculae, slightly thinning of trabeculae and none obvious slight fractures in femurs after 12 weeks of treatment with ZGZTF. The expression of LRP5,β-catenin, Runx2 and Osx involved in the canonical Wnt/β-catenin signaling pathway was significantly up-regulated in the presence of ZGZTF,and the expression of SOST in this pathway was down-regulated. Conclusions These results suggest that ZGZTF may be antiosteoporosis by down-regulating SOST protein to promote Wnt/β-catenin signaling pathway.展开更多
Previous studies by us and others demonstrated that activation of Wnt/β-catenin signaling plays a pathogenic role in chronic kidney diseases(CKD).Wnt co-receptor LRP5 variants are reported to associate with autosomal...Previous studies by us and others demonstrated that activation of Wnt/β-catenin signaling plays a pathogenic role in chronic kidney diseases(CKD).Wnt co-receptor LRP5 variants are reported to associate with autosomal dominant polycystic kidney disease;but their exact roles in this disease and renal fibrosis have not been explored.Here,we observed the upregulation of LRP5 in the renal tubules of both type 1 and type 2 diabetic models and of an obstructive nephropathy model.In the obstructed kidneys,Lrp5 knockout significantly ameliorated tubulointerstitial fibrosis and tubular injury without changing Wnt/β-catenin signaling.Instead,decreased levels of TGF-β1 and TGF-βreceptors(TβRs)were detected in Lrp5 knockout kidneys,followed by attenuated activation and nuclear translocation of Smad2/3 in the renal tubules,suggesting a regulatory effect of LRP5 on TGF-β/Smad signaling.In consistent with this hypothesis,LRP5 overexpression resulted in enhanced TGF-β/Smad signaling activation in renal tubule epithelial cells.Furthermore,LRP5 was co-immunoprecipitated with TβRI and TβRII,and its extracellular domain was essential for interacting with TβRs and for its pro-fibrotic activity.In addition to stabilizing TβRs,LRP5 increased the basal membrane presentation and TGF-β1-induced internalization of these receptors.Notably,TGF-β1 also induced LRP5 internalization.These findings indicate that LRP5 promotes tubulointerstitial fibrosis,at least partially,via direct modulation of TGF-β/Smad signaling,a novel,Wnt-independent function.展开更多
基金the funding support from the National Natural Science Foundation of China (No. 81573956)Natural Science Foundation of Hunan Province (No. 2018JJ2297)+3 种基金Key Program of Chinese Medicine Science Research Plan of Hunan Province (No. 201612)Key Program of Scientific Research Fund of Hunan Provincial Education Department (No. 16A162)National College Students Innovation and Entrepreneurship Project (No. 201710541002)The Project of Research Learning and Innovative Experiment for College Students in Hunan (No. 2015217, No. 2015220, No. 2016284 and No. 2016281)
文摘Objective To observe the effects of Zhuang Gu Zhi Tong Formula (ZGZTF) on antagonist SOST in canonical Wnt/β-catenin signaling pathway in osteoporosis. Methods We analyzed the differential genes of patients with osteoporosis and normal subjects from the GEO database and then found SOST with specific expression. We analyzed SOST as an antagonist of the Wnt signaling pathway by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Then we studied the effect of ZGZTF on SOST in Wnt signaling pathway. Osteoporosis model was induced by ovariectomy (OVX) in 8-week-old female Sprague–Dawley (SD) rats. After 12 weeks of treatment with ZGZTF by intragastric administration, the rats were put to death in batch. The changes of alkaline phosphatase (ALP), bone gla protein (BGP) and estradiol (E2) in serum were determined, and bone mineral density (BMD) and histomorphology of right femur were observed. Biomechanics of lumbar vertebra were measured, and the expression of SOST, Wnt3a,β-catenin, LRP5, Runx2, Osx and their mRNA involving the canonical Wnt/β-catenin signaling pathway were detected by Western blot, RT-PCR and Immunohistochemical analysis. All data were analyzed by SPSS 22.0. Results Twelve weeks of treatment with ZGZTF could significantly decrease the level of ALP and BGP in serum, increase the BMD of femurs, and improve the biomechanical capability of vertebral body in maximum loading and elastic modulus. Concerning histomorphology, we found ordered arrangement of trabeculae, slightly thinning of trabeculae and none obvious slight fractures in femurs after 12 weeks of treatment with ZGZTF. The expression of LRP5,β-catenin, Runx2 and Osx involved in the canonical Wnt/β-catenin signaling pathway was significantly up-regulated in the presence of ZGZTF,and the expression of SOST in this pathway was down-regulated. Conclusions These results suggest that ZGZTF may be antiosteoporosis by down-regulating SOST protein to promote Wnt/β-catenin signaling pathway.
基金supported by a grant from the Oklahoma Center for the Advancement of Science and Technology(HR16-041)and NIH grants EY012231,EY018659,EY028949,EY019309,GM122744a grant from National Nature Science Foundation of China NO.81700631Science&Technology Development Fund of Tianjin Education Commission for Higher Education 2016YD05.
文摘Previous studies by us and others demonstrated that activation of Wnt/β-catenin signaling plays a pathogenic role in chronic kidney diseases(CKD).Wnt co-receptor LRP5 variants are reported to associate with autosomal dominant polycystic kidney disease;but their exact roles in this disease and renal fibrosis have not been explored.Here,we observed the upregulation of LRP5 in the renal tubules of both type 1 and type 2 diabetic models and of an obstructive nephropathy model.In the obstructed kidneys,Lrp5 knockout significantly ameliorated tubulointerstitial fibrosis and tubular injury without changing Wnt/β-catenin signaling.Instead,decreased levels of TGF-β1 and TGF-βreceptors(TβRs)were detected in Lrp5 knockout kidneys,followed by attenuated activation and nuclear translocation of Smad2/3 in the renal tubules,suggesting a regulatory effect of LRP5 on TGF-β/Smad signaling.In consistent with this hypothesis,LRP5 overexpression resulted in enhanced TGF-β/Smad signaling activation in renal tubule epithelial cells.Furthermore,LRP5 was co-immunoprecipitated with TβRI and TβRII,and its extracellular domain was essential for interacting with TβRs and for its pro-fibrotic activity.In addition to stabilizing TβRs,LRP5 increased the basal membrane presentation and TGF-β1-induced internalization of these receptors.Notably,TGF-β1 also induced LRP5 internalization.These findings indicate that LRP5 promotes tubulointerstitial fibrosis,at least partially,via direct modulation of TGF-β/Smad signaling,a novel,Wnt-independent function.
