Abstract: Hypertension is a prevalent systemic disease in the elderly, who can suffer from several pathological skeletal conditions simultaneously, including osteoporosis. Benidipine (BD), which is widely used to treat hypertension, has been proved to have a beneficial effect on bone metabolism. In order to confirm the osteogenic effects of BD, we investigated its osteogenic function using mouse MC3T3-E1 preosteoblast cells in vitro. The proliferative ability of MC3T3-E1 cells was significantly associated with the concentration of BD, as measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cell cycle assay. With BD treatment, the osteogenic differentiation and maturation of MC3T3-E1 cells were increased, as established by the alkaline phosphatase (ALP) activity test, matrix mineralized nodules formation, osteogenic genetic test, and protein expression analyses. Moreover, our data showed that the BMP2/Smad pathway could be the partial mechanism for the promotion of osteogenesis by BD, while BD might suppress the possible function of osteoclasts through the OPG/RANKL/RANK (receptor activator of nuclear factor-κB (NF-κB)) pathway. The hypothesis that BD bears a considerable potential in further research on its dual therapeutic effect on hypertensive patients with poor skeletal conditions was proved within the limitations of the present study.

降压药贝尼地平在体外对小鼠MC3T3-E1细胞促成骨作用的初步研究

[1]AfzalF, PratapJ, ItoK, et al., 2005. Smad function and intranuclear targeting share a Runx2 motif required for osteogenic lineage induction and BMP2 responsive transcription. J Cell Physiol, 204(1):63-72.

[2]BoyleWJ, SimonetWS, LaceyDL, 2003. Osteoclast differentiation and activation. Nature, 423(6937):337-342.

[3]CaoX, ChenD, 2005. The BMP signaling and in vivo bone formation. Gene, 357(1):1-8.

[4]ChenMM, ZhangYH, DuYP, et al., 2019. Epidemiological and clinical study of hip fracture in hospitalized elderly patients in Shanghai, China. Arch Osteoporos, 14:37.

[5]ChenP, LiZZ, HuYH, 2016. Prevalence of osteoporosis in China: a meta-analysis and systematic review. BMC Public Health, 16:1039.

[6]ChenX, WangZQ, DuanN, et al., 2018. Osteoblast‒osteoclast interactions. Connect Tissue Res, 59(2):99-107.

[7]CuiZY, MengXY, FengH, et al., 2019. Estimation and projection about the standardized prevalence of osteoporosis in mainland China. Arch Osteoporos, 15:2.

[8]DucyP, StarbuckM, PriemelM, et al., 1999. A Cbfa1-dependent genetic pathway controls bone formation beyond embryonic development. Genes Dev, 13(8):1025-1036.

[9]GaoSW, LiuF, 2019. Novel insights into cell cycle regulation of cell fate determination. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(6):467-475.

[10]GhoshM, MajumdarSR, 2014. Antihypertensive medications, bone mineral density, and fractures: a review of old cardiac drugs that provides new insights into osteoporosis. Endocrine, 46(3):397-405.

[11]HiguchiA, LingQD, HsuST, et al., 2012. Biomimetic cell culture proteins as extracellular matrices for stem cell differentiation. Chem Rev, 112(8):4507-4540.

[12]HuangZN, RenPG, MaT, et al., 2010. Modulating osteogenesis of mesenchymal stem cells by modifying growth factor availability. Cytokine, 51(3):305-310.

[13]IlićK, ObradovićN, Vujasinović-StuparN, 2013. The relationship among hypertension, antihypertensive medications, and osteoporosis: a narrative review. Calcif Tissue Int, 92(3):217-227.

[14]InayoshiA, SugimotoY, FunahashiJ, et al., 2011. Mechanism underlying the block of human Ca_v3.2 T-type Ca²⁺ channels by benidipine, a dihydropyridine Ca²⁺ channel blocker. Life Sci, 88(19-20):898-907.

[15]KosakaH, HirayamaK, YodaN, et al., 2010. The L-, N-, and T-type triple calcium channel blocker benidipine acts as an antagonist of mineralocorticoid receptor, a member of nuclear receptor family. Eur J Pharmacol, 635(1-3):49-55.

[16]LarijaniB, BekheirniaMR, SoltaniA, et al., 2004. Bone mineral density is related to blood pressure in men. Am J Hum Biol, 16(2):168-171.

[17]LeeKS, HongSH, BaeSC, 2002. Both the Smad and p38 MAPK pathways play a crucial role in Runx2 expression following induction by transforming growth factor-β and bone morphogenetic protein. Oncogene, 21(47):7156-7163.

[18]LimLS, FinkHA, BlackwellT, et al., 2009. Loop diuretic use and rates of hip bone loss and risk of falls and fractures in older women. J Am Geriatr Soc, 57(5):855-862.

[19]LiuL, WangD, QinY, et al., 2019. Astragalin promotes osteoblastic differentiation in MC3T3-E1 cells and bone formation in vivo. Front Endocrinol, 10:228.

[20]LynnH, KwokT, WongSYS, et al., 2006. Angiotensin converting enzyme inhibitor use is associated with higher bone mineral density in elderly Chinese. Bone, 38(4):584-588.

[21]MaJ, WangZ, ZhaoJQ, et al., 2018. Resveratrol attenuates lipopolysaccharides (LPS)‍-induced inhibition of osteoblast differentiation in MC3T3-E1 cells. Med Sci Monit, 24:2045-2052.

[22]MaJY, YouD, LiWY, et al., 2019. Bone morphogenetic proteins and inner ear development. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(2):131-145.

[23]MaZP, LiaoJC, ZhaoC, et al., 2015. Effects of the 1,4-dihydropyridine L-type calcium channel blocker benidipine on bone marrow stromal cells. Cell Tissue Res, 361(2):467-476.

[24]MetzJA, MorrisCD, RobertsLA, et al., 1999. Blood pressure and calcium intake are related to bone density in adult males. Br J Nutr, 81(5):383-388.

[25]MundyGR, ChenD, ZhaoM, et al., 2001. Growth regulatory factors and bone. Rev Endocr Metab Disord, 2(1):105-115.

[26]NeveA, CorradoA, CantatoreFP, 2013. Osteocalcin: skeletal and extra-skeletal effects. J Cell Physiol, 228(6):1149-1153.

[27]NishiyaY, SugimotoS, 2001. Effects of various antihypertensive drugs on the function of osteoblast. Biol Pharm Bull, 24(6):628-633.

[28]NishiyaY, KosakaN, UchiiM, et al., 2002. A potent 1,4-dihydropyridine L-type calcium channel blocker, benidipine, promotes osteoblast differentiation. Calcif Tissue Int, 70(1):30-39.

[29]NoëlD, GazitD, BouquetC, et al., 2004. Short-term BMP-2 expression is sufficient for in vivo osteochondral differentiation of mesenchymal stem cells. Stem Cells, 22(1):74-85.

[30]ShimizuH, NakagamiH, YasumasaN, et al., 2012. Links between hypertension and osteoporosis: benidipine ameliorates osteoporosis in ovariectomized hypertensive rats through promotion of osteoblast proliferation and inhibition of osteoclast differentiation. Curr Cardiovasc Risk Rep, 6(4):274-280.

[31]SimsNA, GooiJH, 2008. Bone remodeling: multiple cellular interactions required for coupling of bone formation and resorption. Semin Cell Dev Biol, 19(5):444-451.

[32]SudoH, KodamaHA, AmagaiY, et al., 1983. In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol, 96(1):191-198.

[33]SuzukiH, YokoyamaK, AkimotoY, et al., 2007. Clinical efficacy of benidipine for vasospastic angina pectoris. Arzneimittelforschung, 57(1):20-25.

[34]TetiA, 2011. Bone development: overview of bone cells and signaling. Curr Osteoporos Rep, 9(4):264-273.

[35]WangBX, BiM, ZhuZ, et al., 2014. Effects of the antihypertensive drug benidipine on osteoblast function in vitro. Exp Ther Med, 7(3):649-653.

[36]WangZW, ChenZ, ZhangLF, et al., 2018. Status of hypertension in China: results from the China hypertension survey, 2012‒2015. Circulation, 137(22):2344-2356.

[37]WuMR, ChenGQ, LiYP, 2016. TGF-β and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Res, 4:16009.

[38]XueC, ZhouCC, YangB, et al., 2017. Comparison of efficacy and safety between benidipine and hydrochlorothiazide in fosinopril-treated hypertensive patients with chronic kidney disease: protocol for a randomised controlled trial. BMJ Open, 7(2):e013672.

[39]YangSM, NguyenND, EismanJA, et al., 2012. Association between beta-blockers and fracture risk: a Bayesian meta-analysis. Bone, 51(5):969-974.

[40]YaoK, NagashimaK, MikiH, 2006. Pharmacological, pharmacokinetic, and clinical properties of benidipine hydrochloride, a novel, long-acting calcium channel blocker. J Pharmacol Sci, 100(4):243-261.

[41]YuD, WangJ, QianKJ, et al., 2020. Effects of nanofibers on mesenchymal stem cells: environmental factors affecting cell adhesion and osteogenic differentiation and their mechanisms. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(11):871-884.