Abstract
Background: (E)-methyl-cinnamate (EMC), a phytochemical constituent isolated from Alpinia katsumadai Hayata, is a natural flavor compound with anti-inflammatory properties, which is widely used in the food and commodity industry. However, the pharmacological effects of methyl-cinnamate on pre-osteoblasts remain unknown. This study aimed to investigate the pharmacological effects and mechanisms of EMC in pre-osteoblast MC3T3-E1 cells (pre-osteoblasts). Methods: Cell viability and apoptosis were evaluated using the MTT assay and TUNEL staining. Cell migration and osteoblast differentiation were examined using migration assays, as well as alkaline phosphatase activity and staining assays. Western blot analysis was used to examine intracellular signaling pathways and apoptotic proteins. Results: EMC decreased cell viability with morphological changes and increased apoptosis in pre-osteoblasts. EMC also induced the cleavage of Poly (ADP-ribose) polymerase (PARP) and caspase-3 and reduced the expression of anti-apoptotic proteins. In addition, EMC increased TUNEL-positive cells in pre-osteoblasts, decreased the activation of mitogen-activated protein kinases, and suppressed cell migration rate in pre-osteoblasts. Subsequently, EMC inhibited the osteoblast differentiation of pre-osteoblasts, as assessed by alkaline phosphatase staining and activity assays. Conclusion: These findings demonstrate that EMC has a pharmacological and biological role in cell survival, migration, and osteoblast differentiation. It suggests that EMC might be a potential phytomedicine for treating abnormalities of osteoblast function in bone diseases.
Highlights
Alpinia katsumadai Hayata is widely used in traditional Chinese medicine to treat emesis and gastric disorders [1]
EMC Decreases Cell Survival and Induces Morphological Changes in Pre-Osteoblasts (E)-methyl-cinnamate (EMC) was isolated from the seeds of A. katsumadai using High-performance liquid chromatography (HPLC) to evaluate its effect o(nE)p-mree-othsytel-ocbinlansatms.aTteh(eEsMtrCu)ctwuraes ainsodlaHtePdLfCrocmhrtohme asteoegdrsamof oAf.EkMatsCumaraedasihuoswinnginHFPiLgCurteo1A,B
Bone pathophysiology is regulated by the proliferation, apoptosis, migration, and differentiation of osBteoonbelpasattsh[o1p1h,1y3s,i1o7l]o.gTyhies rimegpualiartmedenbtyotfhoespteroolbiflaersat tfiuonnc,taiopnopctaoussies,smseivgeraratilobno, naenddidsiefafesreesn, stiuacthioans oofsotesotepoobrloassitss, [p1e1r,i1o3d,1o7n]t.aTlhdeisiemapsea,iromsteenopt oetfroossties,oabnladstPfaugnectt’siodnisceaaussee[s1s4e,1v9e–ra2l1b].oAnepodpisteoassisesis, siunicthiaallsy oisntdeoupceodrobsyis,thpeeraicotdivoanttiaolndoisfecaassep, aossete-8opaentdro-s9ist,harnoudgPhagtheet’sexdtirsienassiec [d1e4a,1th9–r2e1c]e.pAtoproppatothsiws aisyinanitdialtlhye induced by the activation of caspase-8 and -9 through the extrinsic death receptor pathway and the intrinsic mitochondrial pathway, leading to the activation of executioner caspase-3 [22]
Summary
Alpinia katsumadai Hayata is widely used in traditional Chinese medicine to treat emesis and gastric disorders [1]. The seeds of A. katsumadai have been used as an antioxidant and a stomachic [2]. (E)-methyl-cinnamate (EMC), isolated from the seeds of A. katsumadai, is a methyl ester synthesized by cinnamic acid methyltransferase from cinnamic acid. Cinnamic acid is an antioxidant phytochemical agent and is known to be a safe flavoring used in the food industry [4,5]. The following pharmacological properties of EMC have been demonstrated: antibacterial and anti-fungal [6,7]. This includes being antispasmodic and myorelaxant for anti-neuropathic pain and anti-inflammatory effects [8,9]. Sci. 2019, 20, x FOR PEER REVIEW there is no information on the pharmacolog2icoaf l1e4ffects and biological actions of EMC in osteoblasts
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