CAJ | 학술논문

目的观察甲状旁腺激素对去卵巢SD大鼠及其原代成骨细胞基质GLA蛋白(MGP)表达的影响,探讨MGP在绝经后骨质疏松症发病机制中的可能作用.方法 36只SD雌鼠被分为3组,每组12只:假手术组、单纯去卵巢组(OVX组)和去卵巢加甲状旁腺激素干预组(PTH组).PTH组大鼠去卵巢3周后,予甲状旁腺激素(2 μg/kg,每周3次,持续12周).所有大鼠18周后处死.每3周收集尿及血液.观察腰椎组织及骨密度变化,酶联免疫吸附( ELISA)法检测血清及尿液中MGP的量;免疫组化法观察腰椎未羧化MGP的表达,荧光实时定量观察腰椎及成骨细胞MGP mRNA的表达.结果甲状旁腺激素干预后去卵巢SD大鼠血清及尿液中MGP量,腰椎未羧化MGP表达、MGP mRNA表达与单纯去卵巢组比较差异有显著性(P＜0.05),甲状旁腺激素( 1-34)在10-2mol/L、10-8mol/L、10-9mol/L上调成骨细胞MGPmRNA的表达较空白对照组分别为6.78倍、5.31倍和2.23倍,呈剂量依赖性,差异均有统计学意义(P＜0.05).结论甲状旁腺激素对MGP基因及蛋白表达的影响可能在绝经后骨质疏松发病机制中发挥重要作用.
목적 관찰갑상방선격소대거란소SD대서급기원대성골세포기질GLA단백(MGP)표체적영향,탐토MGP재절경후골질소송증발병궤제중적가능작용.방법 36지SD자서피분위3조,매조12지:가수술조、단순거란소조(OVX조)화거란소가갑상방선격소간예조(PTH조).PTH조대서거란소3주후,여갑상방선격소(2 μg/kg,매주3차,지속12주).소유대서18주후처사.매3주수집뇨급혈액.관찰요추조직급골밀도변화,매련면역흡부( ELISA)법검측혈청급뇨액중MGP적량;면역조화법관찰요추미최화MGP적표체,형광실시정량관찰요추급성골세포MGP mRNA적표체.결과 갑상방선격소간예후거란소SD대서혈청급뇨액중MGP량,요추미최화MGP표체、MGP mRNA표체여단순거란소조비교차이유현저성(P＜0.05),갑상방선격소( 1-34)재10-2mol/L、10-8mol/L、10-9mol/L상조성골세포MGPmRNA적표체교공백대조조분별위6.78배、5.31배화2.23배,정제량의뢰성,차이균유통계학의의(P＜0.05).결론 갑상방선격소대MGP기인급단백표체적영향가능재절경후골질소송발병궤제중발휘중요작용.
Objective To observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.Methods Thirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.Conclusion The effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis