Apocynum venetum leaf extract ameliorates postmenopausal osteoporosis by inhibiting RANKL-induced osteoclastogenesis

Abstract

Identifying novel therapeutic efficacies of traditional Chinese medicine (TCM) and their underlying mechanisms is essential for TCM modernization and development. Apocynum venetum leaf (AVL) is used both as an antihypertensive medicine and as a food. Whether AVL extract possesses additional therapeutic efficacies remains to be investigated. Here, we examined the potential effect of AVL extract on ameliorating postmenopausal osteoporosis (PMO) and identified the effective fractions and underlying mechanisms of its efficacy. We found that the total extract of AVL substantially relieved osteoporosis symptoms in both mouse and rat ovariectomized models. Moreover, the AVL-95% fraction, accounting for only about 2% of the total extract, exhibited a therapeutic effect on osteoporosis comparable to that of the total extract. Notably, the AVL-95% fraction improved both cancellous and cortical bones. Additionally, it exerted beneficial effects on both femurs and vertebrae in ovariectomized (OVX) mice. Mechanistic studies showed that AVL-95% inhibited osteoclastogenesis by suppressing RANKL signaling. Importantly, AVL-95% did not exhibit apparent toxicity in acute and chronic toxicity evaluations. Collectively, our work reveals a previously unknown efficacy of AVL in PMO intervention and an underlying mechanism of this efficacy, highlighting the anti-PMO application potential of AVL.

Keywords

Apocynum venetum leaf / AVL fraction / Postmenopausal osteoporosis / Osteoclastogenesis / RANKL

Funding

This work was supported by the National Natural Science Foundation of China (Nos. 32270758, 32070779 and 31770811).

Supporting information

Supporting information for this work can be obtained by contacting the corresponding authors via E-mail.

Declaration of competing interest

These authors have no conflict of interest to declare.

References

[1] Yu H, Jiang L, Wan B, et al. The role of aryl hydrocarbon receptor in bone remodeling. Prog Biophys Mol Biol. 2018; 134:44-49. https://doi.org/10.1016/j.pbiomolbio.2017.12.005.

[2] Li Z, Li D, Chen R, et al. Cell death regulation: a new way for natural products to treat osteoporosis. Pharmacol Res. 2023; 187:106635. https://doi.org/10.1016/j.phrs.2022.106635.

[3] Lu L, Tian L. Postmenopausal osteoporosis coexisting with sarcopenia: the role and mechanisms of estrogen. J Endocrinol. 2023; 259(1):e230116. https://doi.org/10.1530/JOE—23—0116.

[4] Lisco G, Triggiani D, Giagulli VA, et al. Endocrine, metabolic, and immune pathogenesis of postmenopausal osteoporosis: is there a therapeutic role in natural products? Endocr Metab Immune Disord Drug Targets. 2023; 23(10):1278-1290. https://doi.org/10.2174/1871530323666230330121301.

[5] Jiang X, Kagan R. Hormone therapy for postmenopausal osteoporosis management. Climacteric. 2022; 25(1):50-55. https://doi.org/10.1080/13697137.2021.1957818.

[6] Novak S, Roeder E, Kalinowski J, et al. Osteoclasts derive predominantly from bone marrow—resident CX3CR1+ precursor cells in homeostasis, whereas circulating CX3CR1+ cells contribute to osteoclast development during fracture repair. J Immunol. 2020; 204(4):868-878. https://doi.org/10.4049/jimmunol.1900665.

[7] Lacey DL, Boyle WJ, Simonet WS, et al. Bench to bedside: elucidation of the OPG—RANK—RANKL pathway and the development of denosumab. Nat Rev Drug Discov. 2012; 11(5):401-419. https://doi.org/10.1038/nrd3705.

[8] Wang J, Ye T, Wang H, et al. The role of TAK1 in RANKL—induced osteoclastogenesis. Calcif Tissue Int. 2022; 111(1):1-12. https://doi.org/10.1007/s00223—022—00967—z.

[9] Lee K, Chung YH, Ahn H, et al. Selective regulation of MAPK signaling mediates RANKL—dependent osteoclast differentiation. Int J Biol Sci. 2016; 12(2):235-245. https://doi.org/10.7150/ijbs.13814.

[10] Xu Q, Chen G, Liu X, et al. Icariin inhibits RANKL—induced osteoclastogenesis via modulation of the NF—κB and MAPK signaling pathways. Biochem Biophys Res Commun. 2019; 508(3):902-906. https://doi.org/10.1016/j.bbrc.2018.11.201.

[11] Hiligsmann M, Evers SM, Ben Sedrine W, et al. A systematic review of cost—effectiveness analyses of drugs for postmenopausal osteoporosis. Pharmacoeconomics. 2015; 33(3):205-224. https://doi.org/10.1007/s40273—014—0231—1.

[12] Lewiecki EM. Emerging drugs for postmenopausal osteoporosis. Expert Opin Emerg Drugs. 2009; 14(1):129-144. https://doi.org/10.1517/14728210902766813.

[13] Cranney A, Wells GA. Hormone replacement therapy for postmenopausal osteoporosis. Clin Geriatr Med. 2003; 19(2):361-370. https://doi.org/10.1016/S0749—0690(02)00078—2.

[14] Gennari L, Rotatori S, Bianciardi S, et al. Treatment needs and current options for postmenopausal osteoporosis. Expert Opin Pharmacother. 2016; 17(8):1141-1152. https://doi.org/10.1080/14656566.2016.1176147.

[15] Xie W, Zhang X, Wang T, et al. Botany, traditional uses, phytochemistry and pharmacology of Apocynum venetum L. (Luobuma): a review. J Ethnopharmacol. 2012; 141(1):1-8. https://doi.org/10.1016/j.jep.2012.02.003.

[16] Wei JM. Progress in the study on the medical effects of Apocynum venetum (A. lancifolium). J Tradit Chin Med. 1988; 8(1):34-36.

[17] Xiang T, Wu L, Isah MB, et al. Apocynum venetum, a medicinal, economical and ecological plant: a review update. Peer J. 2023; 11:e14966. https://doi.org/10.7717/peerj.14966.

[18] Fukuda T, Takeda S, Xu R, et al. Sema3A regulates bone—mass accrual through sensory innervations. Nature. 2013; 497(7450):490-493. https://doi.org/10.1038/nature12115.

[19] Sato S, Hanada R, Kimura A, et al. Central control of bone remodeling by neuromedin U. Nat Med. 2007; 13(10):1234-1240. https://doi.org/10.1038/nm1640.

[20] Xu R, Zhang C, Shin DY, et al. c—Jun N—terminal kinases (JNKs) are critical mediators of osteoblast activity in vivo. J Bone Miner Res. 2017; 32(9):1811-1815. https://doi.org/10.1002/jbmr.3184.

[21] Tou JC. Resveratrol supplementation affects bone acquisition and osteoporosis: pre—clinical evidence toward translational diet therapy. Biochim Biophys Acta Mol Basis Dis. 2015; 1852(6):1186-1194. https://doi.org/10.1016/j.bbadis.2014.10.003.

[22] Mosekilde L. Assessing bone quality: animal models in preclinical osteoporosis research. Bone. 1995; 17(4 Suppl):343S-352S.

[23] Ma HR, Wang J, Qi HX, et al. Assessment of the estrogenic activities of chickpea (Cicer arietinum L.) sprout isoflavone extract in ovariectomized rats. Acta Pharmacol Sin. 2013; 34(3):380-386. https://doi.org/10.1038/aps.2012.160.

[24] Windahl SH, Andersson G, Gustafsson JA. Elucidation of estrogen receptor function in bone with the use of mouse models. Trends Endocrinol Metab. 2002; 13(5):195-200. https://doi.org/10.1016/S1043—2760(02)00594—5.

[25] Xiang J, Lan R, Tang YP, et al. Apocynum venetum leaf extract attenuates disruption of the blood—brain barrier and upregulation of matrix metalloproteinase—9/—2 in a rat model of cerebral ischemia—reperfusion injury. Neurochem Res. 2012; 37(8):1820-1828.

[26] Wang WQ, Liang XY, Fu D, et al. Apocynum venetum leaf attenuates myocardial ischemia/reperfusion injury by inhibiting oxidative stress. Am J Chin Med. 2015; 43(1):71-85. https://doi.org/10.1142/S0192415X15500056.

[27] Lim SY, Bolster MB. Current approaches to osteoporosis treatment. Curr Opin Rheumatol. 2015; 27(3):216-224. https://doi.org/10.1097/BOR.0000000000000169.

[28] Honig S, Rajapakse CS, Chang G. Current treatment approaches to osteoporosis. Bull Hosp Jt Dis. 2013; 71(3):184-188. https://doi.org/10.1097/bor.0000000000000169.

[29] Sindel D. Osteoporosis: spotlight on current approaches to pharmacological treatment. Turk J Phys Med Rehabil. 2023; 69(2):140-152. https://doi.org/10.5606/tftrd.2023.13054.

[30] Robinson LJ, Yaroslavskiy BB, Griswold RD, et al. Estrogen inhibits RANKL—stimulated osteoclastic differentiation of human monocytes through estrogen and RANKL—regulated interaction of estrogen receptor—alpha with BCAR1 and TRAF6. Exp Cell Res. 2009; 315(7):1287-1301. https://doi.org/10.1016/j.yexcr.2009.01.014.

[31] Chen F, Ouyang Y, Ye T, et al. Estrogen inhibits RANKL—induced osteoclastic differentiation by increasing the expression of TRPV5 channel. J Cell Biochem. 2014; 115(4):651-658. https://doi.org/10.1002/jcb.24700.

[32] Banu J, Varela E, Fernandes G. Alternative therapies for the prevention and treatment of osteoporosis. Nutr Rev. 2012; 70(1):22-40. https://doi.org/10.1111/j.1753—4887.2011.00451.x.

[33] Zhang ND, Han T, Huang BK, et al. Traditional Chinese medicine formulas for the treatment of osteoporosis: implication for antiosteoporotic drug discovery. J Ethnopharmacol. 2016; 189:61-80. https://doi.org/10.1016/j.jep.2016.05.025.

[34] Ai C, Zou Y, Liu H, et al. Traditional Chinese herbal medicine for allergic diseases: a review. Am J Chin Med. 2023; 51(4):779-806. https://doi.org/10.1142/S0192415X23500374.

[35] Zeng P, Zhou H, Guo P, et al. Efficacy and safety of traditional Chinese herbal medicine in the treatment of threatened abortion: a protocol for systematic review and meta—analysis. Medicine (Baltimore). 2021; 100(5):e23288. https://doi.org/10.1097/MD.0000000000023288.

[36] Yamatsu A, Yamashita Y, Maru I, et al. The improvement of sleep by oral intake of GABA and Apocynum venetum leaf extract. J Nutr Sci Vitaminol (Tokyo). 2015; 61(2):182-187. https://doi.org/10.3177/jnsv.61.182.

[37] Ma YX, Chen SY. Observations on the anti—aging, antihypertensive and antihyperlipemic effect of Apocynum venetum leaf extract. Chin J Integr Med. 1989; 9(6):335-337, 323.

[38] Reinhardt RA, Payne JB, Maze CA, et al. Influence of estrogen and osteopenia/osteoporosis on clinical periodontitis in postmenopausal women. J Periodontol. 1999; 70(8):823-828. https://doi.org/10.1902/jop.1999.70.8.823.

[39] Kusumbe AP, Ramasamy SK, Starsichova A, et al. Sample preparation for high—resolution 3D confocal imaging of mouse skeletal tissue. Nat Protoc. 2015; 10(12):1904-1914. https://doi.org/10.1038/nprot.2015.125.