title: Kaempferol targets Krt-14 and induces cytoskeletal mineralization in osteoblasts: A mechanistic approach.
abstract: Kaempferol (KEM) has been observed to stimulate Krt-14 protein which subsequently contributes to matrix maturation and mineralization in rat primary osteoblast cells. Incorporation of Krt-14 siRNA results in reduced mRNA and protein expression after 48h post transfection and remained low for 9days. At day 9 Krt-14 siRNA significantly reduced mineralization without concomitant change in the cell number. ColI and OCN gene expression was reduced in Krt-14 siRNA-treated osteoblast cells. Soluble osteocalcin and collagen levels were markedly decreased in conditioned medium as well as in acid-salt soluble cell-ECM layer treated with Krt-14 siRNA compared to control siRNA treated cells corroborated at the ultrastructral level by AFM. Further, knockdown of Krt-14 and inhibitors against AMPK and mTOR, repressed the activation of mTOR and mineralization attenuated by KEM confirmed the role of Krt-14 in mineralization. These findings strongly suggest that Krt-14 regulates osteoblast mineralization by organizing osteoblast derived ECM.
authors.affiliation: Division of Endocrinology, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Vikram
authors.initials: V
authors.lastname: Khedgikar
authors.affiliation: Division of Endocrinology, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Kushwaha
authors.affiliation: Division of Endocrinology, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Gautam
authors.affiliation: Division of Pharmaceutics, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Shewta
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Division of Pharmaceutics, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Ashwni
authors.initials: A
authors.lastname: Verma
authors.affiliation: Division of Endocrinology, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Dharmendra
authors.initials: D
authors.lastname: Choudhary
authors.affiliation: Division of Pharmaceutics, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India.
authors.forename: Prabhat R
authors.initials: PR
authors.lastname: Mishra
authors.affiliation: Division of Endocrinology, Central Drug Research Institute, (CSIR-CDRI), Lucknow 226031, India. Electronic address: ritu_trivedi@cdri.res.in.
authors.forename: Ritu
authors.initials: R
authors.lastname: Trivedi
mesh_terms: D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects*; D002452:Cell Count; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D055785:Gene Knockdown Techniques; D044949:Kaempferols / Q000494:pharmacology*; D053547:Keratin-14 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism* / Q000648:ultrastructure; D015675:Osteocalcin / Q000378:metabolism; D011720:Pyrazoles / Q000037:antagonists & inhibitors; D011743:Pyrimidines / Q000037:antagonists & inhibitors; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D020123:Sirolimus / Q000037:antagonists & inhibitors; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: D044949:Kaempferols; D053547:Keratin-14; C507539:Krt14 protein, rat; D011720:Pyrazoles; D011743:Pyrimidines; D034741:RNA, Small Interfering; D015675:Osteocalcin; C516138:dorsomorphin; C006552:kaempferol; D003094:Collagen; C546845:mTOR protein, rat; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus
keywords: Atomic force microscopy; Cytokeratin-14; Kaempferol; Mineralization; Osteoblast; mTOR signaling