Medicarpin, a legume phytoalexin, stimulates osteoblast differentiation and promotes peak bone mass achievement in rats: evidence for estrogen receptor β-mediated osteogenic action of medicarpin

Bhargavan, Biju; Singh, Divya; Gautam, Anurag; Mishra, Jay S.; Kumar, Amit; Goel, Atul; Dixit, Manish; Pandey, Rashmi; Manickavasagam, Lakshmi; Dwivedi, Shailendra D.; Chakravarti, Bandana; Jain, Girish Kumar; Ramachandran, Ravishankar; Maurya, Rakesh; Trivedi, Arun Kumar; Chattopadhyay, Naibedya; Sanyal, Sabyasachi

doi:10.1016/j.jnutbio.2010.11.002

Cited by 60 publications

(56 citation statements)

References 54 publications

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“…Our previous studies established the osteogenic efficacy of Med [5]. Med enhanced osteoblast differentiation and mineralization via the stimulation of ER-b/BMP-2-mediated pathway [5].…”

Section: De and Ms Analysis To Identify Protein Differentially Regulmentioning

confidence: 95%

“…Primary culture of osteoblast cells isolated from calvariae of neonatal mice using collagenase and dispase digestion method was carried out using our previously published protocol [5,8]. Briefly, calvariae were subjected to five sequential digestions at 37°C in 0.1 % dispase and 0.1 % collagenase P solution.…”

Section: Calvarial Osteoblast Culturementioning

confidence: 99%

“…Studies by our group established that Med promoted the attainment of peak bone mass in growing rats and also prevented ovariectomy-induced bone loss in BALB/c mice [5,8]. Med stimulated the ER/BMP2 signaling pathway to enhance bone formation [5]. Med was also found to heal cortical bone defect in ovariectomized rats and was almost equivalent in its bone healing capacity to the known anabolic agent, PTH [9].…”

Section: Introductionmentioning

confidence: 99%

“…Out of the many, five compounds were found to contribute majorly to the osteogenic efficacy of B. monosperma. These included methoxyisoflavones like formononetin, isoformononetin, cladrin, cajanin, and the pterocarpan medicarpin [3][4][5][6][7][8][9]. Among these five pure compounds, the most effective osteogenic agent was medicarpin (Med), a natural pterocarpan.…”

Section: Introductionmentioning

confidence: 99%

“…Med was the most effective osteogenic compound on virtue of its dual mode of action where it enhanced osteoblast differentiation and also potently inhibited osteoblast differentiation. Studies by our group established that Med promoted the attainment of peak bone mass in growing rats and also prevented ovariectomy-induced bone loss in BALB/c mice [5,8]. Med stimulated the ER/BMP2 signaling pathway to enhance bone formation [5].…”

Section: Introductionmentioning

confidence: 99%

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Identification of GRP78 as a molecular target of medicarpin in osteoblast cells by proteomics

et al. 2016

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Osteogenic activity was identified in medicarpin (Med), a natural pterocarpan. Further, it was decided to study the differentially regulated protein expression during osteoblast differentiation in the presence of Med. Using 2D proteomic approach, we found that Med treatment to osteoblasts significantly downregulated GRP78, an ER chaperone with anti-apoptotic properties which also controls the activation of unfolded protein response signaling, a pro-survival strategy for normal ER functioning. However, severe stress leads to triggering of apoptotic responses and signaling switches to pro-apoptotic. In order to elucidate the effect of Med downregulation of GRP78, osteoblasts were transfected with SiGRP78 or SiGRP78+ Med or Med alone. It was seen that mRNA and protein levels of ER stress markers like GRP78, ATF-4, and CHOP were decreased in all the three groups with maximum reduction in SiGRP78+ Med group. Med targets GRP78 by inhibiting mitochondrial-mediated apoptosis which is evident by reduced levels of cytochrome c, caspase-3, Bax/BCL2 ratio, and enhanced expression of survivin. Finally, Annexin-PI staining of apoptotic cells revealed that MED inhibition of GRP78 leads to reduced osteoblast apoptosis and increased osteoblast survival. Altogether, our data show that Med inhibits ER stress-induced apoptosis and promotes osteoblast cell survival by targeting GRP78.

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“…Our previous studies established the osteogenic efficacy of Med [5]. Med enhanced osteoblast differentiation and mineralization via the stimulation of ER-b/BMP-2-mediated pathway [5].…”

Section: De and Ms Analysis To Identify Protein Differentially Regulmentioning

confidence: 95%

Section: Calvarial Osteoblast Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of GRP78 as a molecular target of medicarpin in osteoblast cells by proteomics

et al. 2016

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MiR 376c inhibits osteoblastogenesis by targeting Wnt3 and ARF‐GEF‐1 ‐facilitated augmentation of beta‐catenin transactivation

Kureel

John

Prakash

et al. 2017

J of Cellular Biochemistry

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Wnt signaling pathway plays important role in all aspects of skeletal development which include chondrogenesis, osteoblastogenesis, and osteoclastogenesis. Induction of the Wnt-3 signaling pathway promotes bone formation while inactivation of the pathway leads to bone related disorders like osteoporosis. Wnt signaling thus has become a desired target to treat osteogenic disorders. MicroRNAs (miRNAs) represent an important category of elements that interact with Wnt signaling molecules to regulate osteogenesis. Here, we show that miR-376c, a well-characterized tumor suppressor which inhibits cell proliferation and invasion in osteosarcoma by targeting to transforming growth factor-alpha, suppresses osteoblast proliferation, and differentiation. Over-expression of miR-376c inhibited osteoblast differentiation, whereas inhibition of miR-376c function by antimiR-376c promoted expression of osteoblast-specific genes, alkaline phosphatase (ALP) activity, and matrix mineralization. Target prediction analysis tools and experimental validation by luciferase 3' UTR reporter assay along with qRT-PCR identified Wnt-3 and ARF-GEF-1 as direct targets of miR-376c. It was seen that over-expression of miR-376c leads to repression of canonical Wnt/β-catenin signaling. Our overall results suggest that miR-376c targets Wnt-3 and ARF-GEF-1 suppresses ARF-6 activation which prevents the release of β-catenin and its transactivation thereby inhibiting osteoblast differentiation. Although miR-376c is known to be a tumor repressor; we have identified a second complementary function of miR-376c where it inhibits Wnt-3-mediated osteogenesis and promotes bone loss.

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Maintenance of increased bone mass after PTH withdrawal by sequential medicarpin treatment via augmentation of cAMP‐PKA pathway

Sharma

Awasthi

Prakash

et al. 2022

J of Cellular Biochemistry

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Osteoporosis is a metabolic bone disorder associated with impaired bone microarchitecture leading to fragility fractures. Long‐term usage of parathyroid hormone (PTH) enhances bone resorption and leads to osteosarcoma in rats which limits its exposure to maximum 2 years in human. Notably, the anabolic effects of PTH do not endure in the absence of sustained administration. Studies in our lab identified osteogenic and antiresorptive activity in medicarpin, a phytoestrogen belonging to the pterocarpan class. Considering dual‐acting property of medicarpin and limitations of PTH therapy, we envisaged that medicarpin sequential treatment after PTH withdrawal could serve as promising therapeutic approach for osteoporosis treatment. As PTH exerts its bone anabolic effect by increasing osteoblast survival, our study aims to determine whether medicarpin amplifies this effect of PTH. Our results show that PTH withdrawal led to reduced bone mineral density and bone parameters, while sequential treatment of medicarpin after PTH withdrawal significantly enhanced these parameters. Remarkably, these effects were more pronounced than 8‐week PTH treatment. Sequential therapy also significantly increased P1NP levels and decreased CTX levels and TRAP positive cells compared to PTH 8W group where CTX levels were quite high due to bone resorptive action of PTH. Protein expression studies revealed that medicarpin along with PTH betters the antiapoptotic potential compared to PTH alone, through augmentation of cyclic adenosine monophosphate‐PKA‐CREB pathway. These results proclaim that medicarpin sequential treatment prevented the reduction in bone accrual and strength accompanying PTH withdrawal and also aided in antiapoptotic role of PTH. The study points toward the potential use of medicarpin as a replacement therapeutic option postdiscontinuation of PTH.

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Medicarpin, a legume phytoalexin, stimulates osteoblast differentiation and promotes peak bone mass achievement in rats: evidence for estrogen receptor β-mediated osteogenic action of medicarpin

Cited by 60 publications

References 54 publications

Identification of GRP78 as a molecular target of medicarpin in osteoblast cells by proteomics

Identification of GRP78 as a molecular target of medicarpin in osteoblast cells by proteomics

MiR 376c inhibits osteoblastogenesis by targeting Wnt3 and ARF‐GEF‐1 ‐facilitated augmentation of beta‐catenin transactivation

Maintenance of increased bone mass after PTH withdrawal by sequential medicarpin treatment via augmentation of cAMP‐PKA pathway

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