Bone metabolism in a rat model of diabetes mellitus intervened by Xianling Gubao Capsules

doi:10.12307/2022.941

Abstract

Abstract: BACKGROUND: Xianling Gubao Capsules (a commonly used Chinese medicine in orthopedic surgery) can improve bone metabolism and prevent osteoporosis; however, whether it has a preventive effect or a possible mechanism on osteoporosis complicated by diabetes has been rarely reported.
OBJECTIVE: To investigate the effects of Xianling Gubao Capsules on bone metabolism and related osteogenic marker factors Wnt3a, β-catenin and bone morphogenetic protein-2 in rats with diabetes mellitus complicated by osteoporosis and to explore their mechanisms of action.
METHODS: Thirty-six male healthy 8-week-old Sprague-Dawley rats were treated with streptozotocin at a dose of 60 mg/kg to establish a diabetes mellitus model and were then randomly divided into model group, Xianling Gubao Capsule group and alendronate group, with 12 rats in each group. Another 12 normal rats were used as the normal group. Rats in the Xianling Gubao Capsule group and alendronate group were given Xianling Gubao Capsule and alendronate sodium suspension by gavage for 12 weeks, respectively. Rats in the normal and model groups were gavaged with the same amount of double-distilled water for 12 weeks. Blood glucose level of model rats was measured once a week during the administration period to ensure the accuracy of the experiment. Femur, tibia and systemic bone mineral density in each group were measured by dual energy X-ray absorptiometry. Hematoxylin-eosin staining was used to observe the morphological changes of the right femur in each group. Immunohistochemistry and RT-PCR were applied to detect the expression of osteogenic marker factors Wnt3a, β-catenin, and bone morphogenetic protein-2 at protein and mRNA levels.
RESULTS AND CONCLUSION: Compared with the model group, the femur, tibia and systemic bone mineral density values of rats increased significantly in the Xianling Gubao Capsule group (P < 0.05). The mRNA and protein expressions of Wnt3a, β-catenin, and bone morphogenetic protein-2 in bone tissues decreased significantly in the model group (P < 0.05). While compared with the model group, the mRNA and protein expressions of Wnt3a, β-catenin, and bone morphogenetic protein-2 were significantly increased in the Xianling Gubao Capsule group (P < 0.05). In addition, there was no significant difference in the expressions of above-mentioned indicators between the Xianling Gubao Capsule group and alendronate group. These findings confirmed that Xianling Gubao Capsules exert a protective effect on bone metabolism in diabetic rats by the Wnt /β-catenin signaling pathway.

Key words: diabetes, osteoporosis, Wnt/β-catenin, signaling pathway, Xianling Gubao Capsules, rat

CLC Number:

Jiao Yinghua, Bao Kairan, Song Jieqiong, Xing Lei, Cui Lihua, Gao Jingyuan, Qi Ning, Liu Xiangyu. Bone metabolism in a rat model of diabetes mellitus intervened by Xianling Gubao Capsules[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(32): 5173-5178.

Figures/Tables 4

References

[1] ISHTAYA GA, ANABTAWI YM, ZYOUD SH, et al. Osteoporosis knowledge and beliefs in diabetic patients: a cross sectional study from Palestine. BMC Musculoskelet Disord. 2018;19(1):43.
[2] KURRA S, FINK DA, SIRIS ES. Osteoporosis-associated fracture and diabetes. Endocrinol Metab Clin North Am. 2014;43(1):233-243.
[3] CHANDRAN M, TAY D, HUANG XF, et al. The burden of inpatient care for diabetic and non-diabetic patients with osteoporotic hip fractures—does it differ? An analysis of patients recruited into a fracture liaison service in Southeast Asia. Arch Osteoporos. 2018;13(1):27.
[4] WU J, LI W, YE B, et al. The efficacy and safety of Xianling Gubao capsules in the treatment of knee osteoarthritis: A protocol for a randomized, double-blind, controlled trial. Medicine (Baltimore). 2021; 100(36):e27086.
[5] INDRAN IR, LIANG RLZ, MIN TE, et al. Preclinical studies and clinical evaluation of compounds from the genus Epimedium for osteoporosis and bone health. Pharmacol Ther. 2016;162:188-205.
[6] LI WD, YAN CP, WU Y, et al. Osteoblasts proliferation and differentiation stimulating activities of the main components of Fructus Psoraleae corylifoliae. Phytomedicine.2014;21(4):400-405.
[7] NIU YB, LI YH, KONG XH, et al. The beneficial effect of Radix Dipsaci total saponins on bone metabolism in vitro and in vivo and the possible mechanisms of action. Osteoporos Int. 2012;23(11):2649-2660.
[8] XING Y, BI HY, ZHANG QN. Introduction of common Chinese patent medicines for the treatment of osteoporosis. Chin J Osteoporos. 2013; 19(1):83-85.
[9] ZHU HM, QIN L, GARNERO P, et al. The first multicenter and randomized clinical trial of herbal Fufang for treatment of postmenopausal osteoporosis. Osteoporos Int. 2012;23(4):1317-1327.
[10] RAWADI G, ROMAN-ROMAN S. Wnt signalling pathway: a new target for the treatment of osteoporosis. Expert Opin Ther Targets. 2005;9(5): 1063-1077.
[11] MANOLAGAS SC, ALMEIDA M. Gone with the Wnts: β-catenin, T-cell factor, forkhead box O, and oxidative stress in age-dependent diseases of bone, lipid, and glucose metabolism. Mol Endocrinol. 2007;21(11): 2605-2614.
[12] NIE X, WEI X, MA H, et al. The complex role of Wnt ligands in type 2 diabetes mellitus and related complications. J Cell Mol Med. 2021; 25(14):6479-6495.
[13] LI X, LIU D, LI J, et al. Wnt3a involved in the mechanical loading on improvement of bone remodeling and angiogenesis in a postmenopausal osteoporosis mouse model.FASEB J. 2019;33(8):8913-8924.
[14] WANG BW, FANG WJ, SHYU KG. Micro RNA‐145 regulates disabled‐2 and Wnt3a expression in cardiomyocytes under hyperglycaemia. Eur J Clin Invest. 2018;48(1). doi: 10.1111/eci.12867.
[15] CHEN G, DENG C, LI YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci. 2012;8(2):272-288.
[16] ZHANG JM, YU RQ, WU FZ, et al. BMP‑2 alleviates heart failure with type 2 diabetes mellitus and doxorubicin‑induced AC16 cell injury by inhibiting NLRP3 inflammasome-mediated pyroptosis. Exp Ther Med. 2021;22(2):897.
[17] ISLAM MS. Experimental rodent models of type 2 diabetes: a review. Methods Find Exp Clin Pharmacol. 2009;31(4):249-261.
[18] INABA M, TERADA M, KOYAMA H, et al. Influence of high glucose on 1, 25‐dihydroxyvitamin D3‐induced effect on human osteoblast‐like MG‐63 cells.J Bone Miner Res. 1995;10(7):1050-1056.
[19] MACDONALD BT, TAMAI K, HE X. Wnt/β-catenin signaling: components, mechanisms, and diseases. Dev Cell. 2009;17(1):9-26.
[20] KRISHNAN V, BRYANT HU, MACDOUGALD OA.Regulation of bone mass by Wnt signaling.J Clin Invest. 2006;116(5):1202-1209.
[21] BARON R, KNEISSEL M. WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med. 2013;19(2):179-192.
[22] KOBAYASHI Y, UEHARA S, UDAGAWA N, et al. Regulation of bone metabolism by Wnt signals. J Biochem. 2016;159(4):387-392.
[23] INOUE J, IHARA Y, TSUKAMOTO D, et al. Identification of BCL11B as a regulator of adipogenesis. Sci Rep. 2016;6:32750.
[24] FIORENZANO A, PASCALE E, D’ANIELLO C, et al. Cripto is essential to capture mouse epiblast stem cell and human embryonic stem cell pluripotency. Nat Commun. 2016;7:12589.
[25] MALEKI DANA P, SADOUGHI F, MANSOURNIA MA, et al. Targeting Wnt signaling pathway by polyphenols: implication for aging and age-related diseases. Biogerontology. 2021;22(5):479-494.
[26] DAY TF, YANG Y. Wnt and hedgehog signaling pathways in bone development.J Bone Joint Surg Am. 2008;90 Suppl 1:19-24.
[27] KURRA S, FINK DA, SIRIS ES. Osteoporosis-associated fracture and diabetes.Endocrinol Metab Clin North Am. 2014;43(1):233-243.
[28] GARCÍA-MARTÍN A, ROZAS-MORENO P, REYES-GARCÍA R, et al. Circulating levels of sclerostin are increased in patients with type 2 diabetes mellitus.J Clin Endocrinol Metab. 2012;97(1):234-241.
[29] MA R, WANG L, ZHAO B, et al. Diabetes perturbs bone microarchitecture and bone strength through regulation of Sema3A/IGF-1/β-catenin in rats. Cell Physiol Biochem. 2017;41(1):55-66.
[30] XIONG Y, ZHANG Y, XIN N, et al. 1α, 25-Dihydroxyvitamin D3 promotes osteogenesis by promoting Wnt signaling pathway. J Steroid Biochem Mol Biol. 2017;174:153-160.
[31] Willert K, Brown JD, Danenberg E, et al. Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature. 2003; 423(6938):448-452.
[32] Shimizu H, Julius MA, Giarre M, et al. Transformation by Wnt family proteins correlates with regulation of beta-catenin. Cell Growth Differ. 1997;8(12):1349-1358.
[33] RAWADI G, VAYSSIÈRE B, DUNN F, et al. BMP‐2 controls alkaline phosphatase expression and osteoblast mineralization by a Wnt autocrine loop. J Bone Miner Res. 2003;18(10):1842-1853.
[34] CHEN Y, WANG G, MA Z, et al. Adverse effects of high glucose levels on somite and limb development in avian embryos.Food Chem Toxicol. 2014;71:1-9.
[35] MA XY, FENG YF, MA ZS, et al. The promotion of osteointegration under diabetic conditions using chitosan/hydroxyapatite composite coating on porous titanium surfaces. Biomaterials. 2014;35(26):7259-7270.
[36] SHAO X, YANG Y, TAN Z, et al. Amelioration of bone fragility by pulsed electromagnetic fields in type 2 diabetic KK-Ay mice involving Wnt/β-catenin signaling. Am J Physiol Endocrinol Metab. 2021;320(5): E951-E966.
[37] SCHMITT B, RINGE J, HÄUPL T, et al. BMP2 initiates chondrogenic lineage development of adult human mesenchymal stem cells in high-density culture. Differentiation. 2003;71(9-10):567-577.
[38] INTINI G, NYMAN JS. Dkk1 haploinsufficiency requires expression of Bmp2 for bone anabolic activity. Bone. 2015;75:151-160.
[39] RUAN W, XUE Y, ZONG Y, et al. Effect of BMPs and Wnt3a co-expression on the osteogenetic capacity of osteoblasts. Mol Med Rep. 2016;14(5): 4328-4334.
[40] DUMIC-CULE I, PERIC M, KUCKO L, et al. Bone morphogenetic proteins in fracture repair. Int Orthop. 2018;42(11):2619-2626.
[41] LEE EJ, NA W, KANG MK, et al. Hydroxycoumarin scopoletin inhibits bone loss through enhancing induction of bone turnover markers in a mouse model of type 2 diabetes. Biomedicines. 2021;9(6):648.
[42] KARIM K, GIRIBABU N, SALLEH N. Marantodes pumilum Var Alata (Kacip Fatimah) ameliorates derangement in RANK/RANKL/OPG pathway and reduces inflammation and oxidative stress in the bone of estrogen-deficient female rats with type-2 diabetes. Phytomedicine. 2021;91:153677.
[43] SHAO H, WU R, CAO L, et al. Trelagliptin stimulates osteoblastic differentiation by increasing runt-related transcription factor 2 (RUNX2): a therapeutic implication in osteoporosis. Bioengineered. 2021;12(1): 960-968.
[44] DONG C, YANG H, WANG Y, et al. Anagliptin stimulates osteoblastic cell differentiation and mineralization. Biomed Pharmacother. 2020; 129:109796.
[45] XUE S, GAO P, TAN X, et al. Effect of Down-Regulating AK045490 on Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells (BMSCs) Under High Glucose Environment by Targeting Fat Mass and Obesity-Associated (FTO). Journal of Biomaterials and Tissue Engineering. 2020;10(10):1541-1547.
[46] FATCHIYAH F, SETIAWAN B, SASASE T, et al. The amelioration of T2DM rat femoral bone achieved by anti-osteoporosis of caprine CSN1S2 protein through bone morphogenetic protein signaling pathway. Acta Biochim Pol. 2021;68(2):265-275.