葛根素和白藜芦醇对青年大鼠峰值骨量的影响研究

目的:比较葛根素、白藜芦醇单体与二者的复合药物对大鼠峰值骨量的影响。方法 :选取雌性SPF级Wistar大鼠40只,体重109.45～119.44 g,平均115.87 g,大鼠适应3 d后采用随机数字表法将其分为4组,每组10只,分别为空白对照组(CON组,每日灌胃等体积的蒸馏水),葛根素组(PR组,每日灌胃15.4 mg/kg葛根素),白藜芦醇组(RES组,每日灌胃8.4 mg/kg白藜芦醇),复合药物组(P+R组,每日灌胃8.4 mg/kg白藜芦醇+15.4 mg/kg葛根素复合药)。每7 d监测1次大鼠体重,每月检测1次大鼠全身骨密度。3个月后处死全部大鼠。采用双能X线骨密度仪检测股骨和椎骨离体骨密度;采用万能材料试验机分析骨生物力学;采用VG染色分析骨形态计量学;采用ELISA试剂盒检测血清骨代谢指标及用Micro-CT扫描仪扫描股骨微结构。结果:各组大鼠在实验期间的体重变化无统计学意义;骨密度结果显示:与CON组比较,其余3组股骨和椎骨骨密度均显著升高,且R+P组显著高于PR组和RES组。股骨三点弯曲和椎骨压缩试验结果显示:与CON组相比,其余3组股骨和椎骨最大载荷值均显著升高,且P+R组高于PR组和RES组,而各组间弹性模量值无统计学意义;股骨VG染色结果显示:与CON组相比,其余3组骨小梁数目显著增加,分离度明显减低,且R+P组明显优于PR组和RES组,而各组间骨小梁厚度无显著变化。Micro-CT扫描重建图结果显示:与CON相比,其余3组股骨骨小梁结构紧密,数目较多,分离度小,连续性好,且R+P组显著优于RES和PR组;Micro-CT扫描指标结果显示:与CON组相比,其余3组股骨骨小梁数目(trabecular number,Tb.N)、骨小梁厚度(trabecular thickness,Tb.Th)显著增加,骨小梁分离度(trabecular spacing,Tb.Sp)显著降低,且差异具有统计学意义(P0.05),且R+P组显著优于RES和PR组。血清生化指标结果显示:与CON组相比,其余3组血清中骨钙素(osteocalcin,OC)水平均升高,抗酒石酸酸性磷酸酶5b(TRACP 5b)含量均下降,差异具有显著性,且P+R组显著高于PR组、RES组,而TRACP 5b含量无显著变化。结论:葛根素与白藜芦醇单体按1∶1比例组合而成的复合药在提高青年大鼠峰值骨密度和骨质量,增强骨生物力学性能,促进成骨细胞的矿化成熟,抑制破骨细胞骨吸收方面的药效显著优于葛根素和白藜芦醇单体的作用,表明中药复方或复合药将作为预防和治疗骨质疏松的新途径。

Resveratrol and puerarin on peak bone mass in young rats

Objective: To compare effects of resveratrol,puerarin and the compounds on peak bone mass in rats.Methods: Forty SPF Wistar rats weighed 109.45 g to 119.44 g with an average of 115.87 g were selected. After 3 days'' adaption,rats were divided into control group (the same volume of distilled water per day),puerarin group(15.4 mg/kg puerarin daily),resveratrol group (8.4 mg/kg resveratrol daily),compound drug group (daily dose of 8.4 resveratrol added 15.4 mg/kg of puerarin) and 10 in each group. The body weight of the rats was monitored at every 7 days and body bone density was measured at every month. All rats were sacrificed after 3 months. The bone mineral density of femur and vertebrae was detected by dual energy X-ray absorptiometry;bone biomechanics,VG staining was used to analyze bone histomorphometry;ELISA was used to detect serum bone metabolic index and microstructure of femur were scanned with Micro-CT scanner.Results: There were no significant differences in body weight among groups during exoeriment. Bone mineral density results showed BMD of femur and vertebrae in the other three groups were significantly increased,and R+P group was significantly higher than PR group and RES group(P<0.05) by compared with CON group;three-point bending and compression test results showed compared with CON group,other three groups of femoral and vertebral maximum load values were significantly increased,and P+R group was higher than PR group and RES group,but elastic modulus was not statistically significant. Bone histomorphometry showed that number of trabecular bone in other three groups were significantly increased compared with CON group,separation of trabecular bone were significantly reduced,continuity was improved,and R+P group was significantly better than RES and PR group. The results of Micro-CT scan showed that separation of trabecular bone were significantly reduced,continuity were improved in other three groups,and R+P group was significantly better than RES and PR group. The numbers of trabecular bone (Tb.N),trabecular bone thickness (Tb.Th),volume of trabecular bone (BV/TV) in PR group,RES group and R+P group were significantly higher than CON group,but trabecular bone separation (Tb.Sp) was significantly reduced. Serum levels results showed,level of OC in the other three groups were higher than control group(P<0.05),content of TRACP 5b decreased,and level of OC in P+R group was significantly higher than PR group and RES group,content of TRACP 5b was no significant change.Conclusion: Compound of puerarin and resveratrol assigned in a 1:1 ratio could improve bone mineral density and bone mass in young rats,enhance biomechanical properties of bone,promote mineralization and maturation of osteoblasts,inhibit osteoblastic bone resorption,and is better than the role of their respective monomers. The paper showed that traditional Chinese medicine compound medicine will be used as a new way to prevent and treat osteoporosis.