中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2012年
11期
833-838
,共6页
陈雪英%姜醒华%赖晓阳%方向南%杨雅
陳雪英%薑醒華%賴曉暘%方嚮南%楊雅
진설영%강성화%뢰효양%방향남%양아
雌激素类%骨质疏松,绝经后%钙结合蛋白质类%细胞外基质蛋白质类%阉割%大鼠,Sprague-Dawley
雌激素類%骨質疏鬆,絕經後%鈣結閤蛋白質類%細胞外基質蛋白質類%閹割%大鼠,Sprague-Dawley
자격소류%골질소송,절경후%개결합단백질류%세포외기질단백질류%엄할%대서,Sprague-Dawley
Estrogens%Osteoporosis,postmenopausal%Calcium-binding proteins%Extracellular matrix proteins%Castration%Rats,Sprague-Dawley
目的 观察雌二醇对去卵巢SD大鼠基质GLA蛋白(MGP)表达的影响,探讨雌二醇在改善绝经后骨质疏松中的作用.方法 36只10月龄雌性SD大鼠,随机分为去卵巢组、雌二醇组、对照组,每组12只.去卵巢组、雌二醇组大鼠行双侧卵巢切除术,雌二醇组在去卵巢3周后给予苯甲酸雌二醇肌内注射;对照组仅切除卵巢附近的相当于卵巢大小的脂肪组织;各组大鼠每3周留取血清及尿液,至术后15周处死大鼠,取子宫、腰椎组织行HE染色观察病理学变化;双能X线骨密度仪(DEXA)测定大鼠腰椎骨密度;ELISA法检测血清MGP、雌二醇水平及尿液MGP水平;免疫组化法检测腰椎组织中未梭化MGP蛋白的表达,实时荧光定量PCR技术检测腰椎组织中MGP mRNA的表达.结果(1)术后15周,去卵巢组大鼠子宫内膜萎缩,腰椎骨小梁结构破坏最明显.术后15周血清雌二醇水平去卵巢组为(454±66)pmol/L,对照组为(527±77)pmol/L,雌二醇组为(556±80)pmol/L,去卵巢组与其他两组分别比较,差异均有统计学意义(P＜0.05).雌二醇组、去卵巢组、对照组骨密度分别为(0.279 ±0.024)、(0.263±0.030)、(0.295 ±0.024)g/cm2,去卵巢组与其他两组分别比较,差异均有统计学意义(P＜0.01).(2)雌二醇干预后血清及尿液中MGP出现动态变化.去卵巢组及雌二醇组血清MGP水平均下降,术后9周去卵巢组下降至(104 ±6)ng/L,下降最明显,雌二醇组下降至(134±6)ng/L,两组比较,差异有统计学意义(P＜0.05).对照组术前至术后15周尿液中MGP水平无明显变化(P＞0.05);雌二醇组从术后12周起尿液中MGP水平开始上升,至术后15周时为(110.0±3.4)ng/L,去卵巢组为(86.5±2.5)ng/L,两组比较,差异有统计学意义(P＜0.05).(3)免疫组化染色结果显示,去卵巢组腰椎组织中可见未梭化MGP蛋白呈棕色阳性表达,其他各组表达不明显.(4)腰椎组织中MGP mRNA相对表达水平在去卵巢组为1,雌二醇组为0.289±0.260,对照组为0.103 ±0.098,去卵巢组表达水平较其他各组明显增高,分别比较,差异均有统计学意义(P＜0.01).结论 雌二醇可提高去卵巢大鼠MGP mRNA及蛋白的表达水平,可能在改善绝经后骨质疏松中发挥重要作用.
目的 觀察雌二醇對去卵巢SD大鼠基質GLA蛋白(MGP)錶達的影響,探討雌二醇在改善絕經後骨質疏鬆中的作用.方法 36隻10月齡雌性SD大鼠,隨機分為去卵巢組、雌二醇組、對照組,每組12隻.去卵巢組、雌二醇組大鼠行雙側卵巢切除術,雌二醇組在去卵巢3週後給予苯甲痠雌二醇肌內註射;對照組僅切除卵巢附近的相噹于卵巢大小的脂肪組織;各組大鼠每3週留取血清及尿液,至術後15週處死大鼠,取子宮、腰椎組織行HE染色觀察病理學變化;雙能X線骨密度儀(DEXA)測定大鼠腰椎骨密度;ELISA法檢測血清MGP、雌二醇水平及尿液MGP水平;免疫組化法檢測腰椎組織中未梭化MGP蛋白的錶達,實時熒光定量PCR技術檢測腰椎組織中MGP mRNA的錶達.結果(1)術後15週,去卵巢組大鼠子宮內膜萎縮,腰椎骨小樑結構破壞最明顯.術後15週血清雌二醇水平去卵巢組為(454±66)pmol/L,對照組為(527±77)pmol/L,雌二醇組為(556±80)pmol/L,去卵巢組與其他兩組分彆比較,差異均有統計學意義(P＜0.05).雌二醇組、去卵巢組、對照組骨密度分彆為(0.279 ±0.024)、(0.263±0.030)、(0.295 ±0.024)g/cm2,去卵巢組與其他兩組分彆比較,差異均有統計學意義(P＜0.01).(2)雌二醇榦預後血清及尿液中MGP齣現動態變化.去卵巢組及雌二醇組血清MGP水平均下降,術後9週去卵巢組下降至(104 ±6)ng/L,下降最明顯,雌二醇組下降至(134±6)ng/L,兩組比較,差異有統計學意義(P＜0.05).對照組術前至術後15週尿液中MGP水平無明顯變化(P＞0.05);雌二醇組從術後12週起尿液中MGP水平開始上升,至術後15週時為(110.0±3.4)ng/L,去卵巢組為(86.5±2.5)ng/L,兩組比較,差異有統計學意義(P＜0.05).(3)免疫組化染色結果顯示,去卵巢組腰椎組織中可見未梭化MGP蛋白呈棕色暘性錶達,其他各組錶達不明顯.(4)腰椎組織中MGP mRNA相對錶達水平在去卵巢組為1,雌二醇組為0.289±0.260,對照組為0.103 ±0.098,去卵巢組錶達水平較其他各組明顯增高,分彆比較,差異均有統計學意義(P＜0.01).結論 雌二醇可提高去卵巢大鼠MGP mRNA及蛋白的錶達水平,可能在改善絕經後骨質疏鬆中髮揮重要作用.
목적 관찰자이순대거란소SD대서기질GLA단백(MGP)표체적영향,탐토자이순재개선절경후골질소송중적작용.방법 36지10월령자성SD대서,수궤분위거란소조、자이순조、대조조,매조12지.거란소조、자이순조대서행쌍측란소절제술,자이순조재거란소3주후급여분갑산자이순기내주사;대조조부절제란소부근적상당우란소대소적지방조직;각조대서매3주류취혈청급뇨액,지술후15주처사대서,취자궁、요추조직행HE염색관찰병이학변화;쌍능X선골밀도의(DEXA)측정대서요추골밀도;ELISA법검측혈청MGP、자이순수평급뇨액MGP수평;면역조화법검측요추조직중미사화MGP단백적표체,실시형광정량PCR기술검측요추조직중MGP mRNA적표체.결과(1)술후15주,거란소조대서자궁내막위축,요추골소량결구파배최명현.술후15주혈청자이순수평거란소조위(454±66)pmol/L,대조조위(527±77)pmol/L,자이순조위(556±80)pmol/L,거란소조여기타량조분별비교,차이균유통계학의의(P＜0.05).자이순조、거란소조、대조조골밀도분별위(0.279 ±0.024)、(0.263±0.030)、(0.295 ±0.024)g/cm2,거란소조여기타량조분별비교,차이균유통계학의의(P＜0.01).(2)자이순간예후혈청급뇨액중MGP출현동태변화.거란소조급자이순조혈청MGP수평균하강,술후9주거란소조하강지(104 ±6)ng/L,하강최명현,자이순조하강지(134±6)ng/L,량조비교,차이유통계학의의(P＜0.05).대조조술전지술후15주뇨액중MGP수평무명현변화(P＞0.05);자이순조종술후12주기뇨액중MGP수평개시상승,지술후15주시위(110.0±3.4)ng/L,거란소조위(86.5±2.5)ng/L,량조비교,차이유통계학의의(P＜0.05).(3)면역조화염색결과현시,거란소조요추조직중가견미사화MGP단백정종색양성표체,기타각조표체불명현.(4)요추조직중MGP mRNA상대표체수평재거란소조위1,자이순조위0.289±0.260,대조조위0.103 ±0.098,거란소조표체수평교기타각조명현증고,분별비교,차이균유통계학의의(P＜0.01).결론 자이순가제고거란소대서MGP mRNA급단백적표체수평,가능재개선절경후골질소송중발휘중요작용.
Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.
