Pterostilbene-isothiocyanate impedes RANK/TRAF6 interaction to inhibit osteoclastogenesis, promoting osteogenesis in vitro and alleviating glucocorticoid induced osteoporosis in rats

IF 5.3 2区医学 Q1 PHARMACOLOGY & PHARMACY

Biochemical pharmacology Pub Date : 2022-12-01 DOI:10.1016/j.bcp.2022.115284

Viney Kumar , Swati Haldar , Souvik Ghosh , Samrat Chauhan , Abhishek Sharma , Poonam Dhankhar , Amit Kumar , Satish Jaiswal , Saakshi Saini , Sumeet Gupta , Debrupa Lahiri , Partha Roy

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引用次数: 1

Abstract

Prolonged glucocorticoid treatment often leads to glucocorticoid-induced osteoporosis (GIOP), a common iatrogenic complication. This study has explored the anti-osteoporotic potential of semi-synthetic compound, pterostilbene isothiocyanate (PTER-ITC) in GIOP rat model and bone formation potential in vitro. Dysregulated bone-remodelling leads to osteoporosis. PTER-ITC has shown anti-osteoclastogenic activity in vitro. However, its molecular target remains unidentified, which has been explored in this study through in silico and experimental approaches. Alizarin Red S and von-Kossa staining, and alkaline phosphatase (ALP) activity showed the osteogenic differentiation potential of PTER-ITC in pre-osteoblastic mouse MC3T3-E1 and human hFOB 1.19 cells, further, confirmed through the expressions of osteogenic markers at transcriptional (RT-qPCR) and translational (immunoblotting) levels. The anti-osteoclastogenic property of PTER-ITC was confirmed through inhibition of actin ring formation in mouse RAW 264.7 and human THP-1 macrophagic cells. Molecular docking and molecular dynamic simulation showed that PTER-ITC inhibited the crucial osteoclastogenic RANK/TRAF6 interaction, which was further confirmed biochemically through co-immunoprecipitation assay. Osteoporotic bone architecture [validated through scanning electron microscopy (SEM), X-ray radiography, and micro-computed tomography (µ-CT)], physiology (confirmed through compression testing, Young’s modulus and stress versus strain output) and histology (verified through hematoxylin-eosin, Alizarin Red S, von-Kossa and Masson-trichrome staining) of PTER-ITC-treated GIOP female Wistar rats were assuaged. Osteoporotic amelioration through PTER-ITC treatment was further substantiated through serum biomarkers, like, parathyroid hormone (PTH), ALP, calcium (Ca²⁺), Procollagen type I N-terminal propeptide (P1NP), and 25-hydroxy vitamin D. In conclusion, this study identifies the molecular target of PTER-ITC in impeding osteoclastogenesis and facilitating osteogenesis to ameliorate osteoporosis.

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异硫氰酸紫光芪阻碍RANK/TRAF6相互作用抑制破骨细胞生成，促进体外成骨，减轻糖皮质激素诱导的大鼠骨质疏松症

长期的糖皮质激素治疗经常导致糖皮质激素诱导的骨质疏松症(GIOP)，这是一种常见的医源性并发症。本研究探讨了半合成化合物pterostilbene isothiocyanate (PTER-ITC)在GIOP大鼠模型中的抗骨质疏松潜力和体外成骨潜力。骨重塑失调导致骨质疏松。PTER-ITC在体外显示出抗破骨细胞活性。然而，其分子靶点仍未确定，本研究通过计算机和实验方法对其进行了探索。茜素红S和von-Kossa染色，碱性磷酸酶(ALP)活性显示PTER-ITC在成骨前小鼠MC3T3-E1和人hFOB 1.19细胞中的成骨分化潜力，进一步通过转录(RT-qPCR)和翻译(免疫印迹)水平上成骨标志物的表达证实。通过抑制小鼠RAW 264.7和人THP-1巨噬细胞中肌动蛋白环的形成，证实了PTER-ITC的抗破骨性。分子对接和分子动力学模拟表明，PTER-ITC抑制破骨细胞关键的RANK/TRAF6相互作用，并通过共免疫沉淀实验进一步生化证实。经pter - tc处理的GIOP雌性Wistar大鼠骨质疏松性骨结构[通过扫描电镜(SEM)、x射线摄影和微计算机断层扫描(µ-CT)验证]、生理学(通过压缩测试、杨氏模量和应力与应变输出证实)和组织学(通过苏木精-伊红、茜素红S、von-Kossa和masson三色染色证实)均得到缓解。通过甲状旁腺激素(PTH)、ALP、钙(Ca2+)、I型前胶原n端前肽(P1NP)、25-羟基维生素d等血清生物标志物进一步证实了PTER-ITC治疗骨质疏松的改善作用。综上所述，本研究确定了PTER-ITC阻碍破骨细胞生成、促进成骨以改善骨质疏松的分子靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemical pharmacology 医学-药学

CiteScore

10.30

自引率

1.70%

发文量

420

审稿时长

17 days

期刊介绍： Biochemical Pharmacology publishes original research findings, Commentaries and review articles related to the elucidation of cellular and tissue function(s) at the biochemical and molecular levels, the modification of cellular phenotype(s) by genetic, transcriptional/translational or drug/compound-induced modifications, as well as the pharmacodynamics and pharmacokinetics of xenobiotics and drugs, the latter including both small molecules and biologics. The journal''s target audience includes scientists engaged in the identification and study of the mechanisms of action of xenobiotics, biologics and drugs and in the drug discovery and development process. All areas of cellular biology and cellular, tissue/organ and whole animal pharmacology fall within the scope of the journal. Drug classes covered include anti-infectives, anti-inflammatory agents, chemotherapeutics, cardiovascular, endocrinological, immunological, metabolic, neurological and psychiatric drugs, as well as research on drug metabolism and kinetics. While medicinal chemistry is a topic of complimentary interest, manuscripts in this area must contain sufficient biological data to characterize pharmacologically the compounds reported. Submissions describing work focused predominately on chemical synthesis and molecular modeling will not be considered for review. While particular emphasis is placed on reporting the results of molecular and biochemical studies, research involving the use of tissue and animal models of human pathophysiology and toxicology is of interest to the extent that it helps define drug mechanisms of action, safety and efficacy.