白藜芦醇复合新黄芪茯苓对抗大鼠高原骨质疏松的作用

EFFECT OF RESVERATROL COMBINED WITH NEW HUANGQI-FULING ON OSTEOPOROSIS IN RATS EXPOSED TO HIGH ALTITUDE

目的研究白藜芦醇复合新黄芪茯苓对高原骨代谢紊乱的影响。方法雄性Wistar大鼠36只随机分为常氧对照组(0.5%羧甲基纤维素钠,n=12)、缺氧组(模拟海拔6000米20 h/d,0.5%羧甲基纤维素钠,n=12)及缺氧复合制剂干预组(模拟海拔6000米20 h/d,400mg/(kg·d)白藜芦醇及新黄芪茯苓干预9w,n=12)。低压缺氧干预8w后,使用双能X射线骨密度仪检测大鼠股骨骨密度。于低压缺氧9w时实施动物安乐死,并采集骨组织及血清样本。采用双抗体一步夹心法酶联免疫吸附试验检测大鼠血清中骨钙素含量;使用黄嘌呤氧化酶法测定血清超氧化物歧化酶水平;使用TBA法检测血清丙二醛水平及ABTS法测定血清总抗氧化能力,使用HE染色和免疫组织化学染色于光镜下观察骨组织显微结构及破骨分化情况。结果与常氧对照组相比,缺氧组骨密度下降约15%(P<0.01),缺氧复合制剂组较缺氧组,骨密度明显升高(P<0.05)。与常氧对照组相比,缺氧组大鼠血清骨钙素、超氧化物歧化酶、总抗氧化能力下降(P<0.05),血清丙二醛含量显著升高(P<0.05),与缺氧组相比,缺氧复合制剂干预组血清骨钙素、超氧化物歧化酶、总抗氧化能力升高(P<0.05),血清丙二醛含量下降(P<0.01)。与常氧对照组相比,缺氧组大鼠骨小梁排列紊乱、稀疏,破骨分化标志物TRAP染色区域明显增多,与缺氧组相比,缺氧复合制剂组骨小梁排列及分布趋于正常,破骨分化标志物显著减少。结论白藜芦醇和新黄芪茯苓组成的新型复合制剂可纠正机体骨质代谢紊乱。白藜芦醇和新黄芪茯苓组成的新型复合制剂为抗氧化制剂,本文观察的结果提示抗氧化制剂可能对低氧所致骨代谢紊乱具有积极的改善作用,其具体机制尚需进一步探讨。

Objective To explore the therapeutic effect of resveratrol combined with new Huangqi-Fuling on bone metabolism disorders in rats exposed to simulated high-altitude hypoxia.Methods Thirty-six male Wistar rats were randomly divided into the normal group(0.5%sodium carboxymethyl cellulose,n=12),hypoxia group(a simulated altitude of 6000 meters 20 h/d,0.5%sodium carboxymethyl cellulose,n=12)and hypoxia compound preparation intervention group(a simulated altitude of 6000 meters 20 h/d,400 mg/(kg·d)resveratrol and new Huangqi-Fuling intervention for 9 weeks,n=12).After 8 weeks of low-pressure hypoxia exposure,the dual-energy X-ray bone densitometer was used to detect the bone mineral density of the rats’femur.The rats were euthanized after 9 weeks of hypobaric hypoxia exposure before bone tissue and serum samples were collected.The double-antibody one-step sandwich enzyme-linked immunosorbent assay was used to detect the content of osteocalcin(OCN)in the serum.The xanthine oxidase method was used to determine the serum superoxide dismutase(SOD)activity.The serum malondialdehyde(MDA)was determined using the TBA method and the total antioxidant capacity(T-AOC)using the ABTS method.Results The BMD of the hypoxia group was significantly decreased by approximately 15%(P<0.01)compared with the normal control group.The BMD of the compound group was significantly increased(P<0.05)compared with hypoxia group.Compared with the normal control group,the content of serum OCN and the activity of SOD,T-AOC were decreased(P<0.05),but the level of MDA was increased significantly(P<0.05)in the hypoxia group.Compared with the hypoxia group,the content of serum OCN and the activity of SOD,T-AOC were increased(P<0.05),but the level of MDA was decreased(P<0.01)after intragastric administration of compound preparation.Compared with the normal group,the bone rabecula arrangement of the rats in the hypoxia group was disordered and sparse,and the osteoclast differentiation marker TRAP staining area was significantly increased.Compared with the hypoxia group,the arrangement and distribution of bone trabecula in the compound intervention group tended to be recovered,and osteoclast differentiation markers were significantly reduced.Conclusion The new compound preparation composed of resveratrol and new Huangqi-Fuling is an antioxidant preparation,which can correct bone metabolism disorders induced by hypoxia.The specific mechanism needs to be further explored.