右归饮联合盐酸雷洛昔芬给药治疗小鼠骨质疏松症

摘要

Objective To observe the effect of Yougui Drink (YGD) and raloxifene hydrochloride (RH) combined administration on mice with osteoporosis for investigating the mechanisms,so as to provide experimental basis for the treatment of osteoporosis.Methods The osteoporosis models were simulated by removal of the ovary in C57BL/6 mice.The mice were randomly divide into four groups after 12 weeks of surgery:model,YGD (0.1 g/per mouse),RH (10 mg/kg) and combined administration (YGD 0.1 g/per mouse and RH 10 mg/kg),and sham-operated group.All the mice were killed after 7 weeks of daily ig administration.Bone mineral density (BMD) and the bone microtomographic histomorphometry were assessed by Micro-CT,and the bone histomorphometry index:bone volume/total volume (BV/TV),bone surface area/bone volume (BS/BV),trabecular thickness (Tb.Th),trabecular number (Tb.N),and trabecular separation (Tb.Sp) were detected.Next,the levels of P1NP and CTx in serum were assessed.Finally,the isolated bone mesenchymal stem cells (BMSCs) were cultured for 7 d in vitro for assessing the cell proliferation by CCK-8 method and the ability of osteogenic differentiation by quantative analysis of alkaline phosphatase (ALP).Results Compared with model group,BMD,BV/TV,Tb.Th and Tb.N of YGD group,RH group and combined administration group significantly increased,BS/BV and Tb.Sp decreased significantly (P ＜ 0.05);Compared with RH group,BV/TV,Tb.Th,Tb.N in combined treatment group increased significantly,BS/BV and Tb.Sp decreased significantly (P ＜ 0.05).The P 1NP level and cell proliferation rate of the combined drug group were much higher than those in the other groups (P ＜ 0.05,0.01).The ALP level was much higher than that in sham group,model group and RH group,but much lower than that of YGD group (P ＜ 0.01).Conclusion YGD combined with RH were more effective to attenuate bone loss of the mice than single dose of YGD and RH,by promoting cell proliferation and differentiation to improve the ability of bone formation,thus increasing the bone density and the treatment effect of osteoporosis.%目的 观察右归饮和盐酸雷洛昔芬联合给药对骨质疏松模型小鼠的治疗作用,并研究其作用机制.方法 将C57BL/6雌性小鼠切除卵巢制备骨质疏松模型,手术12周后将小鼠随机分为4组:模型组、右归饮(0.1g/只)组、盐酸雷洛昔芬(10mg/kg)组和联合给药(右归饮0.1g/只与盐酸雷洛昔芬10 mg/kg联合给药)组,另设假手术组,连续7周每天ig给药1次.给药结束后,将动物处死,Micro-CT测定骨密度和骨组织微形态,并测定骨形态计量指标——骨体积分数(BV/TV)、骨表面积/骨体积(B S/BV)、骨小梁的平均厚度(Tb.Th)、骨小梁数量(Tb.N)、骨小梁分离度(Tb.Sp);试剂盒法测定血清中Ⅰ型胶原氨基端(P1NP)、胶原羧基端(CTx)水平;分离并培养骨髓间充质干细胞,体外培养7d,CCK-8法测定细胞增殖率,试剂盒法检测分化相关指标碱性磷酸酶(ALP)水平.结果 与模型组比较,右归饮组、盐酸雷洛昔芬组和联合给药组的骨密度、BV/TV、Tb.Th、Tb.N显著升高,BS/BV和Tb.Sp显著降低(P＜0.05);与盐酸雷洛昔芬组比较,联合给药组的BV/TV、Tb.Th、Tb.N显著升高,BS/BV和Tb.Sp显著降低(P＜0.05).联合给药组P1NP水平、细胞增殖率均显著高于其他各组(P＜0.05、0.01).ALP水平显著高于假手术组、模型组、盐酸雷洛昔芬组,显著低于右归饮组(P＜0.01).结论 在给药时间相同的前提下,右归饮和盐酸雷洛昔芬联合给药比单独给药更有效地抑制小鼠骨丢失,通过促进细胞的增殖和分化提高骨形成能力,达到增加骨密度、治疗骨质疏松的效果.