牛膝多糖对老年骨质疏松大鼠模型骨代谢及生物力学特征的影响
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摘要
目的探讨牛膝多糖对老年骨质疏松大鼠模型骨代谢及生物力学特征的影响。方法将120只10月龄雌性SD大鼠随机分为6组,除对照组和假手术组,均以双侧卵巢摘除术制备骨质疏松模型。术后饲养3个月,低、中、高剂量组以100、200、400 mg/(kg·d)的牛膝多糖灌胃。采用ELISA实验测腹腔静脉血骨钙素(OC)、骨特异性碱性磷酸酶(BAP)、抗酒石酸酸性磷酸酶(TPACP5b)、Ⅰ型胶原交联N-末端肽(NTX)与Ⅰ型胶原交联C-末端肽(CTX)水平;WB实验测定股骨与胫骨骨髓组织核因子κB受体活化因子(RANK)、RANK配体(RANK ligand,RANKL)及骨保护素(OPG)表达;液压伺服材料试验机测定椎体及股骨的最大应力和断裂吸收能量。结果与对照组、假手术组相比,模型组大鼠的血清OC和BAP含量,骨髓组织OPG和RANK表达降低,血清TPACP5b、NTX、CTX含量,骨髓组织RANKL表达升高;与模型组相比,低剂量组、中剂量组、高剂量组大鼠血清OC和BAP含量,骨髓组织OPG和RANK表达依次升高,血清TPACP5b、NTX、CTX含量,骨髓组织RANKL表达依次降低,组间差异有统计学意义(P<0.05);与对照组、假手术组相比,模型组大鼠的椎体和双侧股骨的最大应力及断裂吸收能量明显降低;与模型组相比,低剂量组、中剂量组、高剂量组大鼠的最大应力及断裂吸收能量依次升高;组间差异有统计学意义(P<0.05)。结论牛膝多糖能够改善老年骨质疏松大鼠的骨代谢,提高骨密度和骨抗压能力,在抗骨质疏松领域具有潜在的应用价值。
Objective To explore the influence of achyranthan on bone metabolism and biomechanical characteristics of elderly osteoporosis model rats. Methods Totally 120 female SD rats were divided into 6 groups. In addition to the rats in the control group(Control) and the sham operation group(SO), the others were prepared for osteoporosis modes by surgical castration. After the surgery, the rats in intervention 1, 2, 3 were given intragastric administration with achyranthan of 100, 200, 400 mg/(kg·d), for 3 months. The levels of osteocalcin(OC), bone specific alkaline phosphatase(BAP), tartrate-resistant acid phosphatase(TPACP5 b), Ⅰ type collagen cross-linking N-terminal peptide(NTX) and Ⅰ type collagen cross-linking C-terminal peptide(CTX) of abdominal venous blood of rats were detected by ELISA kits. The receptor activator of NF-kappa b(RANK), RANK ligand(RANKL) and osteoprotegerin(OPG) expression of myelosis of rats were detected by WB. Maximum stress and fracture absorb energy of centrum and thighbone of rats was detected by using servo-hydraulic materials testing machines. Results Compared with the Control/SO group, the serum OC and BAP level, and the medullispinal OPG and RANK expression of the model group decreased, and the serum TPACP5 b, NTX and CTX level, and the medullispinal RANKL expression of the model group increased. Compared with model group, the serum OC and BAP level, and the medullispinal OPG and RANK expression of intervention 1, 2, 3 group decreased successively, and the serum TPACP5 b, NTX and CTX level, and the medullispinal RANKL expression of intervention 1, 2, 3 group increased successively, the difference among the different groups was statistically significant(P<0.05). Compared with the control/SO group, the maximum stress and fracture absorb energy of the model group decreased. Compared with model group, the maximum stress and fracture absorb energy of the intervention 1, 2, 3 group increased successively, the difference among the groups was statistically significant(P<0.05). Conclusion Achyranthan can improve bone metabolism, increase bone density and bone compressive ability of elderly osteoporosis rats, and has potential application value in the field of anti-osteoporosis.
Objective To explore the influence of achyranthan on bone metabolism and biomechanical characteristics of elderly osteoporosis model rats. Methods Totally 120 female SD rats were divided into 6 groups. In addition to the rats in the control group(Control) and the sham operation group(SO), the others were prepared for osteoporosis modes by surgical castration. After the surgery, the rats in intervention 1, 2, 3 were given intragastric administration with achyranthan of 100, 200, 400 mg/(kg·d), for 3 months. The levels of osteocalcin(OC), bone specific alkaline phosphatase(BAP), tartrate-resistant acid phosphatase(TPACP5 b), Ⅰ type collagen cross-linking N-terminal peptide(NTX) and Ⅰ type collagen cross-linking C-terminal peptide(CTX) of abdominal venous blood of rats were detected by ELISA kits. The receptor activator of NF-kappa b(RANK), RANK ligand(RANKL) and osteoprotegerin(OPG) expression of myelosis of rats were detected by WB. Maximum stress and fracture absorb energy of centrum and thighbone of rats was detected by using servo-hydraulic materials testing machines. Results Compared with the Control/SO group, the serum OC and BAP level, and the medullispinal OPG and RANK expression of the model group decreased, and the serum TPACP5 b, NTX and CTX level, and the medullispinal RANKL expression of the model group increased. Compared with model group, the serum OC and BAP level, and the medullispinal OPG and RANK expression of intervention 1, 2, 3 group decreased successively, and the serum TPACP5 b, NTX and CTX level, and the medullispinal RANKL expression of intervention 1, 2, 3 group increased successively, the difference among the different groups was statistically significant(P<0.05). Compared with the control/SO group, the maximum stress and fracture absorb energy of the model group decreased. Compared with model group, the maximum stress and fracture absorb energy of the intervention 1, 2, 3 group increased successively, the difference among the groups was statistically significant(P<0.05). Conclusion Achyranthan can improve bone metabolism, increase bone density and bone compressive ability of elderly osteoporosis rats, and has potential application value in the field of anti-osteoporosis.
引文
[1] 中华医学会骨质疏松和骨矿盐疾病分会.原发性骨质疏松症诊疗指南(2017)[J].中国实用内科杂志,2018,38(2):127-150.
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[10] 黄象维,丁陈颖,徐霞,等.骨代谢标志物与糖尿病患者骨质疏松症的关系[J].中国卫生检验杂志,2018,28(2):206-208.
[11] DOUSTIMOTLAGH A H,DEHPOUR A R,ETEMAD-MOGHADAM S,et al.A study on OPG/RANK/RANKL axis in osteoporotic bile duct-ligated rats and the involvement of nitrergic and opioidergic systems[J].Res Pharm Sci,2018,13(3):239-249.
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[13] 庄海娜,钱婧,杨泽冠,等.基于文献分析的中医药改善骨质疏松症生物力学研究[J].中医学报,2018,33(9):1690-1695.
[14] 张瑶,张维丽,赵筱雯,等.不同补肾方对去卵巢骨质疏松模型大鼠生物力学的影响[J].中国医院药学杂志,2016,36(16):1379-1382.
[15] 齐兵,戴维群,尤建宇,等.不同种药物干预骨质疏松模型大鼠股骨压缩力学的变化[J].中国组织工程研究,2015,19(42):6770-6775.
[2] 胡婷婷,张振凌.中药牛膝化学成分、药理作用及储藏保管[J].中国老年学杂志,2016,36(13):3321-3322.
[3] 王昌盛.怀牛膝糖聚物的分离纯化、结构鉴定及生物活性测试[D].广州:广东药科大学,2016.
[4] 周叶,高越,王晓楠,等.牛膝多糖对硝普钠诱导的兔关节软骨细胞凋亡的影响及机制研究[J].浙江医学,2018,40(7):688-692.
[5] WENG X,LIN P,LIU F,et al.Achyranthes bidentata polysaccharides activate the Wnt/β-catenin signaling pathway to promote chondrocyte proliferation[J].Int J Mol Med,2014,34(4):1045-1050.
[6] 楼家晖,杨鸫祥.牛膝及其复方治疗骨质疏松症临床及实验进展[J].辽宁中医药大学学报,2017,19(10):160-161.
[7] ZHANG M,WANG Y,ZHANG Q,et al.The genus Achyranthes:A review on traditional uses,phytochemistry,and pharmacological activities[J].J Ethnopharmacol,2017,203(1):260-278.
[8] 张萌萌,毛未贤,马倩倩,等.骨代谢标志物在骨质疏松诊疗中的应用指南(2012年版)(日本骨质疏松症学会制定)[J].中国骨质疏松杂志,2013,19(7):645-657.
[9] 巩玲俊.鲑鱼降钙素联合阿仑膦酸钠维D3片及钙剂治疗老年骨质疏松的临床效果观察[J].健康研究,2016,36(6):696-697,699.
[10] 黄象维,丁陈颖,徐霞,等.骨代谢标志物与糖尿病患者骨质疏松症的关系[J].中国卫生检验杂志,2018,28(2):206-208.
[11] DOUSTIMOTLAGH A H,DEHPOUR A R,ETEMAD-MOGHADAM S,et al.A study on OPG/RANK/RANKL axis in osteoporotic bile duct-ligated rats and the involvement of nitrergic and opioidergic systems[J].Res Pharm Sci,2018,13(3):239-249.
[12] SINGH M M,KUMAR R,TEWARI S,et al.Investigation of OPG/RANK/RANKL Genes as a Genetic Marker for Cardiac abnormalities in Thalassemia Major Patients[J].Ann Hum Genet,2017,81(3):117-124.
[13] 庄海娜,钱婧,杨泽冠,等.基于文献分析的中医药改善骨质疏松症生物力学研究[J].中医学报,2018,33(9):1690-1695.
[14] 张瑶,张维丽,赵筱雯,等.不同补肾方对去卵巢骨质疏松模型大鼠生物力学的影响[J].中国医院药学杂志,2016,36(16):1379-1382.
[15] 齐兵,戴维群,尤建宇,等.不同种药物干预骨质疏松模型大鼠股骨压缩力学的变化[J].中国组织工程研究,2015,19(42):6770-6775.