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郭志斌,吴春芳,张国彬,迟博婧,王玉丹,刘子洪,王宝,马超,田发明.辛伐他汀联合利塞膦酸钠对地塞米松诱导大鼠骨丢失的作用[J].中国实验动物学报,2020,28(6):742~748.
辛伐他汀联合利塞膦酸钠对地塞米松诱导大鼠骨丢失的作用
Effects of risedronate sodium combined with simvastatin on dexamethasone-induced bone loss in rats
投稿时间:2020-05-17  
DOI:10. 3969 / j.issn.1005-4847. 2020. 06. 002
中文关键词:  辛伐他汀  利塞膦酸钠  骨丢失  生物力学  微结构
英文关键词:simvastatin  risedronate  bone loss  biomechanics  microarchitecture Conflicts of Interest: The authors declare no conflict of interest
基金项目:
作者单位E-mail
郭志斌 开滦总医院林西医院骨外科,河北 唐山 063000 guozhibin1980@ 126.com 
吴春芳 开滦总医院林西医院骨外科,河北 唐山 063000  
张国彬 华北理工大学,医学实验研究中心, 河北 唐山 063000  
迟博婧 华北理工大学,医学实验研究中心, 河北 唐山 063000  
王玉丹 华北理工大学,医学实验研究中心, 河北 唐山 063000  
刘子洪 开滦总医院骨外科,河北 唐山 063000  
王宝 开滦总医院林西医院骨外科,河北 唐山 063000  
马超 开滦总医院骨外科,河北 唐山 063000  
田发明 华北理工大学,医学实验研究中心, 河北 唐山 063000 tfm9911316@ 163.com 
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中文摘要:
       目的 分析辛伐他汀和利塞膦酸钠单独使用和联合使用对地塞米松诱导的大鼠骨质疏松的作用。 方法 本研究使用雌性 3 月龄 SD 大鼠,分为五组(每组 8 只):对照组、骨质疏松模型组、辛伐他汀组、利塞膦酸钠 组和联合用药组。除对照组外,其余各组大鼠皮下注射地塞米松(2. 5 mg,每周 2 次),药物干预组分别给予辛伐他 汀 5 mg / kg、利塞膦酸钠 0. 1 mg / kg 和两者联合干预,均为每天 1 次,8 周后处死所有大鼠取材。 取左侧股骨检测其 骨密度后,采用三点弯曲实验分析其最大载荷,取左侧胫骨经微计算机断层扫描技术(micro-CT)检测近端松质骨骨量及微观结构参数。取右侧股骨脱钙石蜡包埋,采用免疫组织化学染色检测 OPG 和 RANKL 的表达,提取右侧胫骨 RNA,检测骨保护素(OPG) 和破骨细胞分化因子(RANKL) 的表达。 结果 ( 1) 骨密度:模型组( 0. 207 ± 0. 02) g / cmP>2P>> 显著低于其余各组,辛伐他汀组(0. 226 ± 0. 023) g / cmP>2P>> 、利塞膦酸钠组(0. 237 ± 0. 021) g / cmP>2P>> 显著低于对照组(0. 282 ± 0. 013) g / cmP>2P>> 、联合用药组(0. 257 ± 0. 012) g / cmP>2P>> (P< 0. 05)。 (2)生物力学:最大载荷:模型组(90. 2 ± 7. 1)显著低于对照组和联合用药组(P< 0. 05)。 (3)micro-CT:模型组骨小梁数量(Tb.N)、骨小梁体积分数(BV/ TV)显著低于其他 4 组,骨小梁分离度( Tb. Sp) 显著高于其他 4 组(P< 0. 05),辛伐他汀组( 21. 6 ± 2. 8)、利塞膦酸钠组(21. 9 ± 2. 6)BV/ TV 显著低于对照组(29. 5 ± 2. 7)、联合用药组(25. 3 ± 2. 9),Tb.Sp 显著高于对照组(P< 0. 05)。 (4)免疫组化染色:OPG:模型组、利塞膦酸钠组、联合用药组均显著低于对照组(P< 0. 05),C 组和 E 组显著高于模型组(P< 0. 05)。 模型组、辛伐他汀组、利塞膦酸钠组、联合用药组 RANKL 的表达均显著高于对照组(P< 0. 05),E 组显著低于模型组(P< 0. 05)。 (5)PCR:模型组(0. 74 ± 0. 17)OPG 表达显著低于对照组 (1. 00 ± 0. 16)、辛伐他汀组(1. 27 ± 0. 19) (P< 0. 05);RNAKL 在模型组表达显著高于其余各组(P< 0. 05),辛伐他汀组(1. 31 ± 0. 16)显著高于对照组(1. 00 ± 0. 18)。 结论 地塞米松可诱发建立大鼠骨丢失模型,辛伐他汀和利塞膦酸钠联合应用治疗可部分阻止该模型骨丢失和骨质量下降,效果优于单独用药。
英文摘要:
       Objective To analyze the effects of simvastatin and risedronate sodium, either alone or in combination, on bone loss induced by dexamethasone in rats. Methods Twelve-week-old female Sprague-Dawley rats were divided into five groups ( n = 8 ) as follows: control group, osteoporosis model group subcutaneously injected with dexamethasone (2. 5 mg, twice / week), simvastatin group (5 mg / ( kg·d)), risedronate group (0. 1 mg / ( kg·d)), and combination group. Eight weeks later, all rats were sacrificed, and the left femurs were harvested for bone mineral density (BMD) and three-point bending testing. In addition, micro-CT was performed on the left tibia. The right femurs were embedded in paraffin for immunohistochemical analysis of OPG and RANKL expression, and the right tibia samples were subjected to RNA extraction to assess OPG and RANKL expression. Results BMD was lowest in the osteoporosis group (0. 207 ± 0. 02) g / cmP>2P>> , and BMD was markedly lower in the simvastatin ( 0. 226 ± 0. 023) g / cmP>2P>> and risedronate (0. 237 ± 0. 021) g / cmP>2P>> groups than in the control (0. 282 ± 0. 013) g / cmP>2P>> and combination groups (0. 257 ± 0. 012) g / cmP>2P>> . The maximum load in the osteoporosis group was significantly lower than that in the control and combination groups. BV/ TV and Tb.N were significantly lower in the osteoporosis group than in the other groups, whereas the opposite trend was noted for Tp.Sp. BV/ TV was significantly lower in the simvastatin (21. 6 ± 2. 8) and risedronate groups (21. 9 ± 2. 6) than in the control (29. 5 ± 2. 7) and combination groups (25. 3 ± 2. 9). Tb.Sp was significantly higher in the simvastatin and risedronate groups than in the control group. On immunohistochemistry, OPG expression was significantly lower in the osteoporosis, risedronate, and combination groups than in the control group, whereas its expression was significantly higher in the simvastatin and combination groups than in the osteoporosis group. RANKL expression was significantly higher in the osteoporosis, simvastatin, risedronate, and combination groups than in the control group and significantly lower in the combination group than in the osteoporosis group. Meanwhile, OPG expression was significantly lower in the osteoporosis group (0. 74 ± 0. 17) than in the control (1. 00 ± 0. 16) and simvastatin groups (1. 27 ± 0. 19). RANKL expression was significantly higher in the osteoporosis group than in the other groups, and its expression was significantly higher in the simvastatin group (1. 31 ± 0. 16) than in the control group (1. 00 ± 0. 18). Conclusions Dexamethasone induced bone loss in rats, and this effect was more obviously prevented by combined treatment with simvastatin and risedronate than by either drug alone.
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