摘要
目的 研究淫羊藿苷是否能够预防模拟微重力引起的大鼠骨丢失及氧化应激并探讨其作用机制,为开发源自中草药的预防太空微重力环境骨丢失药物奠定基础.方法 2 月龄Wistar雄性大鼠 30 只,随机分为对照组(CON)、尾吊组(HLS)和淫羊藿苷组(ICA)3 组,每组 10 只.HLS组和ICA组均建立尾吊模型,其中ICA组灌服 50 mg/kg淫羊藿苷,CON组和HLS组只给予等体积蒸馏水.给药28d后处死所有大鼠,左侧股骨和腰椎置于Mirco-CT扫描仪中进行骨微结构分析和三维重建;通过大鼠腹主动脉抽取动脉血,分离血清后用ELISA法测定PINP、CTX-1、OPG、RANKL、8-iso-PGF2α、8-OHdG等的含量;提取右侧胫骨总蛋白,Western blotting法分析BMP2、COL-1、OSX、Runx2的表达水平;TBA法检测血清中的MDA含量,WST-1 法检测血清中的SOD活力.结果 ①Mirco-CT扫描分析结果:与CON组相比,HLS组的骨小梁分布稀疏,而ICA组明显优于HLS组,与CON组接近.骨小梁参数量化情况为:与CON组相比,HLS组的BMD、BS/TV、BV/TV、Tb.N及Tb.Th均降低(P＜0.05),而Tb.Sp升高(P＜0.05).ICA组的BMD、BS/TV、BV/TV、Tb.N及Tb.Th均显著高于HLS组(P＜0.05),而Tb.Sp显著降低(P＜0.05).第4腰椎骨小梁及其量化结果与股骨一致.②血清骨代谢指标的变化情况:HLS组的骨形成指标PINP 和OPG均显著低于CON组(P＜0.05),而骨吸收指标CTX-1和RANKL均高于CON组(P＜0.05);ICA组PINP、OPG含量均显著高于HLS组(P＜0.05),CTX-1、RANKL含量均显著低于HLS组(P＜0.05),其中PINP、OPG和CTX-1含量与CON组水平相近.③胫骨骨组织中HLS组的骨形成蛋白BMP2、Runx2、COL-1和OSX表达水平均低于CON组.ICA组BMP2、Runx2、COL-1、OSX表达水平均高于HLS组,其中Runx2 和OSX表达水平与CON组相近.④氧化与抗氧化指标检测结果:与CON组相比,HLS组血清中 8-iso-PGF2α、8-OHdG、MDA含量升高(P＜0.05),SOD酶活力降低(P＜0.05).与 HLS 组相比,ICA 组血清中 8-iso-PGF2α、8-OHdG、MDA含量降低(P＜0.05),SOD酶活力升高(P＜0.05).结论 模拟微重力环境会引起骨密度下降,最大载荷下降,骨微结构退化,而淫羊藿苷能促进骨形成,抑制骨吸收,减轻体内氧化程度,提升抗氧化能力从而有效预防大鼠产生氧化应激反应导致骨丢失.
Abstract
Objective To study whether icariin can antagonize bone loss induced by simulated microgravity in rats and to explore its mechanism,so as to lay a foundation for the development of anti-space microgravity bone loss drugs derived from Chinese herbal medicine.Methods Thirty 2-month-old Wistar male rats were randomly divided into three groups:control group(CON),tail-hanger group(HLS),and icariside group(ICA),with 10 rats in each group.The tail-hanger model was established in both HLS and ICA groups.Rats in ICA group received 50 mg/kg icariside,while rats in CON and HLS groups received an equal volume of distilled water only.After 28 days of administration,all rats were executed.The left femurs and lumbar vertebrae were placed in a micro-CT scanner for bone microstructure analysis and 3D reconstruction.The arterial blood was extracted from the abdominal aorta of rats.The serum was separated and the contents of PINP,CTX-1,OPG,RANKL,8-iso-PGF2α,and 8-OHdG were determined using ELISA.The total protein was extracted from the right tibia.The expression levels of BMP2,COL-1,OSX,and Runx2 were analyzed using Western blotting method.The MDA content in serum was measured with TBA method.The SOD activity in serum was measured with WST-1 method.Results(1)The result of micro-CT scan analysis showed that compared to the CON group,the distribution of bone trabeculae was sparse in the HLS group,while the ICA group was significantly better than the HLS group and close to the CON group.BMD,BS/TV,BV/TV,Tb.N,and Tb.Th were lower(P＜0.05)and Tb.Sp was higher(P＜0.05)in the HLS group compared to the CON group.BMD,BS/TV,BV/TV,Tb.N,and Tb.Th were significantly higher(P＜0.05)in the ICA group compared to the HLS group,while Tb.Sp was significantly lower(P＜0.05).The trabeculae of the fourth lumbar vertebra and their quantification result were consistent with those of the femur.(2)The changes of serum bone metabolic indexes showed that the bone formation indexes PINP and OPG were significantly lower in the HLS group than in the CON group(P＜0.05),while the bone resorption indexes CTX-1 and RANKL were higher than in the CON group(P＜0.05).The levels of PINP and OPG were significantly higher in the ICA group than in the HLS group(P＜0.05),CTX-1 and RANKL were significantly lower than those in the HLS group(P＜0.05),with the levels of PINP,OPG and CTX-1 similar to those in the CON group.(3)The expression levels of bone formation proteins BMP2,Runx2,COL-1,and OSX in tibial bone tissue were lower in the HLS group than in the CON group.The expression levels of BMP2,Runx2,COL-1,and OSX were higher in the ICA group than in the HLS group,with Runx2 and OSX expression levels similar to those in the CON group.(4)The result of oxidative and antioxidant indexes showed that compared to the CON group,the serum levels of 8-iso-PGF2α,8-OHdG,and MDA increased(P＜0.05)and the SOD enzyme activity decreased(P＜0.05)in the HLS group.The serum levels of 8-iso-PGF2α,8-OHdG,and MDA decreased(P＜0.05)andthe SODenzyme activity increased(P＜0.05)in the ICA group compared to the HLS group.Conclusion The simulated microgravity environment causes a decrease in bone mineral density,a decrease in maximum load,and a degradation of bone microstructure.Icariin promotes bone formation,inhibits bone resorption,reduces the degree of oxidation in vivo,and enhances antioxidant capacity thus effectively preventing bone loss due to oxidative stress in rats.
基金项目
国家自然科学基金面上项目(81770879)
联勤保障部队第九四○医院实验室培育项目(2021yxky081)
甘肃省自然科学基金(22JR11RA013)
NETL
NSTL
维普
万方数据