左归丸对去势骨质疏松大鼠钙磷代谢和细胞核因子κB受体活化因子配体/骨保护素的影响

摘要

目的 探讨左归丸对卵巢摘除大鼠钙磷代谢和细胞核因子κB受体活化因子配体 (receptor acti-vator of nuclear factor κB ligand, RANKL) /骨保护素 (osteoprotegerin, OPG) 的影响.方法 SD雌性大鼠60只, 采用去卵巢法制备骨质疏松症大鼠模型, 将大鼠分为假手术组, 切除卵巢组, 左归丸低、中、高剂量组,西药对照组 (戊酸雌二醇片), 每组10只.造模成功12 周后给予药物干预, 西药对照组给予戊酸雌二醇片0. 09 mg/kg灌胃治疗, 左归丸低、中、高剂量组按4. 75、 9. 5、 19 g/kg灌胃治疗, 假手术组和切除卵巢组大鼠给予等体积蒸馏水灌胃, 干预4 周后股动脉采血处死大鼠.比色分析法检测血清钙 (calcium, Ca)、磷(phosphorus, P) 含量; ELISA法检测骨保护素 (osteoprotegerin, OPG)、细胞核因子κB受体活化因子 (re-ceptor activator of nuclear factor κB, RANK ) 和 RANKL含量; 双能 X骨密度仪分析测定大鼠股骨的骨密度(bone mineral density, BMD); AG-IX生物力学万能实验机检测最大载荷和弹性模量, 比较各组间差异.结果 假手术组, 切除卵巢组, 西药对照组, 左归丸高、中、低剂量组的BMD分别为 (0. 145±0. 004)、 (0. 116± 0. 007)、 (0. 146±0. 006)、 (0. 142±0. 004)、 (0. 134±0. 002)、 (0. 129±0. 001) g/cm2; 这6组的体质量分别为(173. 20±13. 92)、 (264. 10±28. 61)、 (249. 44±26. 64)、 (243. 00±10. 25)、 (253. 45±18. 19)、 (268. 50± 22. 39) g; 这6组的子宫指数分别为 (1. 62±0. 34)、 (0. 45±0. 09)、 (0. 89±0. 22)、 (1. 03±0. 23)、 (0. 91± 0. 13)、 (0. 46±0. 07) mg/g; 这6组Ca、 P含量分别为 (2. 07±0. 07/0. 30±0. 02)、 (0. 92±0. 08/0. 10±0. 03)、(1. 89±0. 11/0. 22±0. 03)、 (1. 95±0. 13/0. 27±0. 02)、 (1. 67±0. 10/0. 19±0. 06)、 (1. 59±0. 15/0. 12±0. 02) mmol/L; 这6组OPG、 RANKL和RANK含量分别为 (755. 00±58. 05/5. 73±0. 27/10. 68±0. 69)、 (493. 67± 36. 77/10. 24± 0. 33/18. 62 ±0. 74)、 (653. 00±56. 93/7. 37±0. 19/12. 23±0. 76)、 (731. 00±38. 49/6. 77±0. 65/11. 52±0. 39)、 (704. 33±62. 92/7. 97±0. 41/14. 69±0. 86)、 (555. 00±97. 09/9. 31±0. 75/15. 04± 0. 52) pg/mL;这6组的最大载荷分别为 (142. 16±0. 10)、 (86. 26±0. 13)、 (126. 43±0. 31)、 (119. 77±0. 74)、 (111. 96± 1. 57)、 (98. 92±1. 44) N; 这6组弹性模量分别为 (19. 48±0. 24)、 (13. 10±0. 43)、 (16. 70±0. 37)、 (16. 17± 0. 15)、 (15. 99±0. 22)、 (14. 98±0. 15) Mpa.与假手术组比较, 切除卵巢组大鼠的BMD、 Ca、 P、 OPG、最大载荷和弹性模量水平均显著降低, 而RANK和RANKL含量均明显升高, 差异有统计学意义 (P<0. 05); 与切除卵巢组相比, 左归丸各剂量干预组和西药对照组大鼠BMD和弹性模量均显著升高, RANK和RANKL含量明显降低, 左归丸中、高剂量组和切除卵巢+西药对照组Ca、 P、 OPG含量和最大载荷显著升高, 差异均有统计学意义 (P<0. 05).结论 左归丸能升高去卵巢大鼠的骨密度, 提高骨强度, 并改善其力学性能, 其机制可能与调节 RANKL/OPG水平和Ca、 P代谢有关.%Objective To investigate the effects of Zuoguiwan treatment on calcium, phosphate metabolism and receptor activator of nuclear factor κB ligand (RANKL) /osteoprotegerin (OPG) in osteoporotic rats induced by ovariec-tomy (OVX). Methods Sixty female SD rats were randomly separated into sham operation group, OVX group, OVX+Zuoguiwan high, medium, low dose group, OVX+western medicine control group ( estradiol valerate tablets), with 10 rats in each group. The OVX model was successful preparation for 12 weeks. OVX+Western medicine control group was given estradiol valerate tablets 0. 09 mg/kg by gavage. OVX+Zuoguiwan low, medium and high dose group was given Zuoguiwan (4. 75, 9. 5, 19 g/kg) by gavage. Sham group and OVX group were given distilled water. The rats were ga-vaged with corresponding drugs for 4 weeks. And colorimetric analysis was used to check the contents of Ca and P. ELISA kits were used to check the contents of OPG, RANK, and RANKL. Dual energy X ray absorptiometer was used to meas-ure bone mineral density (BMD). AG-IX biomechanical universal testing machine was used to measure maximum load and elasticity modulusr. Results BMD in sham operation group, OVX group, OVX+western medicine control group, OVX+Zuoguiwan high, medium, low dose group were (0. 145±0. 004), (0. 116±0. 007), (0. 146±0. 006), (0. 142± 0. 004), (0. 134±0. 002), (0. 129±0. 001) g/cm2, respectively. Body weight in above groups were (173. 20± 13. 92), (264. 10±28. 61), (249. 44±26. 64), (243. 00±10. 25), (253. 45±18. 19), (268. 50±22. 39) g, re-spectively. The uterus index in above groups were (1. 62±0. 34), (0. 45±0. 09), (0. 89±0. 22), (1. 03±0. 23), (0. 91±0. 13), (0. 46±0. 07) mg/g, respectively. Ca, P cotents in above groups were (2. 07±0. 07/0. 30±0. 02), (0. 92±0. 08/0. 10±0. 03), (1. 89±0. 11/0. 22±0. 03), (1. 95±0. 13/0. 27±0. 02), (1. 67±0. 10/0. 19±0. 06), (1. 59±0. 15/0. 12±0. 02) mmol/L, respectively. OPG, RANKL and RANK in above groups were (755. 00±58. 05/5. 73±0. 27/10. 68±0. 69), (493. 67±36. 77/10. 24±0. 33/18. 62±0. 74), (653. 00±56. 93/7. 37±0. 19/12. 23± 0. 76), (731. 00±38. 49/6. 77±0. 65/11. 52 ±0. 39), (704. 33±62. 92/7. 97±0. 41/14. 69±0. 86), (555. 00±97. 09/9. 31±0. 75/15. 04± 0. 52) pg/mL, respectively. Maximum load in above groups were (142. 16±0. 10), (86. 26± 0. 13), (126. 43±0. 31), (119. 77±0. 74), (111. 96±1. 57), (98. 92±1. 44) N, respectively. Elasticity modulus in above groups were (19. 48±0. 24), (13. 10±0. 43), (16. 70±0. 37), (16. 17±0. 15), (15. 99±0. 22), (14. 98± 0. 15) MPa, respectively. Compared with sham group, the levels of BMD, Ca, P, OPG, maximum load, and elasticity modulus were all declined obviously in OVX group ( P<0. 05), but the contents of RANK and RANKL were markedly elevated (P<0. 05). Compared with OVX group, the BMD, elasticity modulus were all increased sharply in OVX+Zouguiwan group and OVX+Western medicine control group (P<0. 05). The Ca, P, OPG, maximum load were also significantly increased in OVX+Zuoguiwan middle, high dose group and OVX+ western medicine control group (P<0. 05). The contents of RANK and RANKL were markedly decreased in ovx+zuoguiwan treatment group and OVX+ west-ern medicine control group (P<0. 05). Conclusion Zuoguiwan is effective in treating ovariectomy induced osteoporosis in rats and the mechanism is mainly achieved by inhibiting bone loss of osteoclasts through regulating the ratio of RANKL and OPG and the metabolism of calcium and phosphorus.