补肾壮骨方对去卵巢大鼠骨结构和骨代谢作用的研究
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摘要
目的研究补肾壮骨方对骨质疏松大鼠骨结构和骨代谢的影响及其可能的作用机制。方法用SD雌性大鼠复制去卵巢骨质疏松模型,然后分别灌服补肾壮骨方水提液高剂量(1.4 g/kg)和低剂量(0.7 g/kg)12周。用生化法测定血清钙(S-Ca)、血清磷(S-P)、尿钙(U-Ca/Cr),尿磷(U-P/Cr)以及血清中高密度脂蛋白(high-density lipoprotein,HDL)、低密度脂蛋白(low-density lipoprotein,LDL)、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)的含量;Elisa法测定I型胶原氨基端前肽(procollagen I N-terminal propeptide,PINP)、I型胶原羧基端肽(collagen I carboxyl terminal peptide,CTX-I)、尿脱氧吡啶啉(deoxypyridine,DPD)、骨钙素(osteocalcin,OCN)的含量;用放免法测定血清碱性磷酸酶(alkaline phosphatase,ALP)的活性;用双能X线(DEXA)法、Viva CT和万能试验机测定骨密度、骨微结构和骨生物力学特性;用Western blot法测定骨组织蛋白酶(cathepsin)K表达。结果补肾壮骨方水提液能升高血清中的钙、磷、HDL-C和PINP的含量,降低血清中TC、TG、LDL、ALP、OCN和CTX-I及尿液中的钙、磷、DPD的含量。同时,补肾壮骨方水提液可以升高去卵巢大鼠股骨的骨密度,改善骨微结构,增加骨强度,降低胫骨Cathepsin K的表达水平。结论补肾壮骨方水提液能够抑制去卵巢大鼠的骨量丢失和骨强度下降,改善去卵巢大鼠的骨代谢,其作用机制可能与抑制Cathepsin K的表达有关。
Objective To investigate the anti-osteoporotic effects of Bushen Zhuanggu compound on bone structure and bone metabolism in ovariectomized(OVX) rats. Methods Osteoporotic rats model was established by bilaterally ovariectomy in Sprague-Dawley female rats. Bushen Zhuanggu compound aqueous extract(0.7 and 1.4 g/kg) were orally given to OVX rats for 12 weeks. Biochemical method was used to determine serum levels of calcium(S-Ca) and phosphorus(S-P), urinary levels of calcium(U-Ca/Cr) and phosphorus(U-P/Cr) as well as serum levels of high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein(LDL), total cholesterol(TC) and triglyceride(TG). ELISA was used to examine collagen I amino terminal peptide(PINP), collagen I carboxyl terminal peptide(CTX-I), osteocalcin(OCN) and urine deoxypyridine(DPD). The activity of serum alkaline phosphatase(ALP) was determined by radioimmunoassay. Western-blotting method was used to measure the expression of cathepsin K in the rat tibia. Bone mineral density, microstructure and biomechanics were determined by dual-energy X-ray(DEXA), VivaCT and bone biomechanical properties. Results Bushen Zhuanggu compound increased the serum levels of calcium, phosphorus, HDL-C and PINP, decreased the urinary levels of calcium and phosphorus and DPD, and reduced serum levels of TC, TG, LDL, ALP, OCN and CTX-I. At the same time, Bushen Zhuanggu Compound could increase the femur bone density, improve bone microstructure, increase bone strength and reduced the expression of tibia cathepsin K in OVX rats. Conclusion Bushen Zhuanggu Compound could exert anti-osteoporosis effect by increasing bone mineral density and strength and improving bone microstructure through regulation of bone metabolism and mineral content in OVX rats, and this effect might be related to inhibition of the cathepsin K expression in rat tibia.
Objective To investigate the anti-osteoporotic effects of Bushen Zhuanggu compound on bone structure and bone metabolism in ovariectomized(OVX) rats. Methods Osteoporotic rats model was established by bilaterally ovariectomy in Sprague-Dawley female rats. Bushen Zhuanggu compound aqueous extract(0.7 and 1.4 g/kg) were orally given to OVX rats for 12 weeks. Biochemical method was used to determine serum levels of calcium(S-Ca) and phosphorus(S-P), urinary levels of calcium(U-Ca/Cr) and phosphorus(U-P/Cr) as well as serum levels of high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein(LDL), total cholesterol(TC) and triglyceride(TG). ELISA was used to examine collagen I amino terminal peptide(PINP), collagen I carboxyl terminal peptide(CTX-I), osteocalcin(OCN) and urine deoxypyridine(DPD). The activity of serum alkaline phosphatase(ALP) was determined by radioimmunoassay. Western-blotting method was used to measure the expression of cathepsin K in the rat tibia. Bone mineral density, microstructure and biomechanics were determined by dual-energy X-ray(DEXA), VivaCT and bone biomechanical properties. Results Bushen Zhuanggu compound increased the serum levels of calcium, phosphorus, HDL-C and PINP, decreased the urinary levels of calcium and phosphorus and DPD, and reduced serum levels of TC, TG, LDL, ALP, OCN and CTX-I. At the same time, Bushen Zhuanggu Compound could increase the femur bone density, improve bone microstructure, increase bone strength and reduced the expression of tibia cathepsin K in OVX rats. Conclusion Bushen Zhuanggu Compound could exert anti-osteoporosis effect by increasing bone mineral density and strength and improving bone microstructure through regulation of bone metabolism and mineral content in OVX rats, and this effect might be related to inhibition of the cathepsin K expression in rat tibia.
引文
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[7] 王丽丽,马如风,于娜,等.丹酚酸B通过抗氧化作用改善高脂饮食小鼠牙槽骨骨质疏松的实验研究[J].中国骨质疏松杂志,2017,23(3):281-285,317.
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[11] 张晓,张国庆,顾伯林,等.丹参及其有效成分对骨代谢影响的实验研究进展[J].中国骨质疏松杂志,2015,21(1):112-116.
[12] Liu C, Wang L, Zhu R, et al. Rehmanniae radix preparata suppresses bone loss and increases bonestrength through interfering with canonical Wnt/β-catenin signaling pathway in OVX rats[J]. Osteoporos Int, 2018 Aug 27. doi: 10.1007/s00198-018-4670-y.
[13] 李建国,谢兴文,李鼎鹏,等.中药淫羊藿治疗骨质疏症的研究进展[J].中国骨质疏松杂志,2018,24(3):389-393.
[14] 卢建华,卢烨超,金红婷,等.红曲对去卵巢大鼠骨质疏松模型血清ALP,TRAP及骨组织TNF-α,RANK表达的影响[J].中国中医骨伤科杂志,2017,25(9):5-8.
[15] 章明放,黄凤歧,谭郁彬,等.去势大鼠血清骨钙素水平及意义[J].中国骨质疏松杂志,1999,5(3):25-27.
[16] Yuan H, Xiao L, Min W, et al. Bu-Shen-Tong-Luo decoction prevents bone loss via inhibition of bone resorption and enhancement of angiogenesis in ovariectomy-induced osteoporosis of rats[J]. J Ethnopharmacol,2018,220:228-238.
[17] Sun P, Liu Y, Deng X, et al. An inhibitor of cathepsin K, icariin suppresses cartilage and bone degradation in mice of collagen-induced arthritis[J]. Phytomedicine,2013,20(11):975-979.
[18] Panwar P,Law S,Jamroz A,et al.Tanshinones: A novel class of ectosteric anti-resorptives[J]. Br J Pharmacol,2018,175(6):902-923.
[2] 白璧辉,谢兴文,李鼎鹏,等.我国近5年来骨质疏松症流行病学研究现状[J].中国骨质疏松杂志,2018,24(2):253-258.
[3] 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.
[4] 孙亚奇,卞艳芳,周春娜,等.中药治疗骨质疏松症研究进展[J].中国临床医学,2018,25(2):307-313.
[5] 郭鱼波,王丽丽,马如风,等.骨质疏松的中医病因病机分析及其中医药治疗的前景探讨[J].世界科学技术-中医药现代化,2015,17(4):768-772.
[6] 郭鱼波,王丽丽,马如风,等.女贞子水提液对去卵巢大鼠骨结构和骨代谢的影响研究[J]. 中草药,2016,47(7):1155-1162.
[7] 王丽丽,马如风,于娜,等.丹酚酸B通过抗氧化作用改善高脂饮食小鼠牙槽骨骨质疏松的实验研究[J].中国骨质疏松杂志,2017,23(3):281-285,317.
[8] 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[J]. Cell Physiol Biochem,2017,41(1):55-66.
[9] Tuazon M, Campbell S, Klein D, et al. Effects of ovariectomy and exercise training intensity on energy substrate and hepatic lipid metabolism, and spontaneous physical activity in mice[J]. Metabolism,2018,83:234-244.
[10] 董世健,杨博文,刘晨,等.从脾肾论治绝经后骨质疏松症的研究进展[J].光明中医,2018,33(17):2620-2622.
[11] 张晓,张国庆,顾伯林,等.丹参及其有效成分对骨代谢影响的实验研究进展[J].中国骨质疏松杂志,2015,21(1):112-116.
[12] Liu C, Wang L, Zhu R, et al. Rehmanniae radix preparata suppresses bone loss and increases bonestrength through interfering with canonical Wnt/β-catenin signaling pathway in OVX rats[J]. Osteoporos Int, 2018 Aug 27. doi: 10.1007/s00198-018-4670-y.
[13] 李建国,谢兴文,李鼎鹏,等.中药淫羊藿治疗骨质疏症的研究进展[J].中国骨质疏松杂志,2018,24(3):389-393.
[14] 卢建华,卢烨超,金红婷,等.红曲对去卵巢大鼠骨质疏松模型血清ALP,TRAP及骨组织TNF-α,RANK表达的影响[J].中国中医骨伤科杂志,2017,25(9):5-8.
[15] 章明放,黄凤歧,谭郁彬,等.去势大鼠血清骨钙素水平及意义[J].中国骨质疏松杂志,1999,5(3):25-27.
[16] Yuan H, Xiao L, Min W, et al. Bu-Shen-Tong-Luo decoction prevents bone loss via inhibition of bone resorption and enhancement of angiogenesis in ovariectomy-induced osteoporosis of rats[J]. J Ethnopharmacol,2018,220:228-238.
[17] Sun P, Liu Y, Deng X, et al. An inhibitor of cathepsin K, icariin suppresses cartilage and bone degradation in mice of collagen-induced arthritis[J]. Phytomedicine,2013,20(11):975-979.
[18] Panwar P,Law S,Jamroz A,et al.Tanshinones: A novel class of ectosteric anti-resorptives[J]. Br J Pharmacol,2018,175(6):902-923.