中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2011年
9期
674-677
,共4页
王玉东%郎虓%陶敏芳%程蔚蔚
王玉東%郎虓%陶敏芳%程蔚蔚
왕옥동%랑효%도민방%정위위
去氢表雄酮%雌激素受体β%成骨细胞
去氫錶雄酮%雌激素受體β%成骨細胞
거경표웅동%자격소수체β%성골세포
Dehydroepiandrosterone%Estrogen receptor beta%Osteoblasts
目的：明确Erβ介导脱氢表雄酮(DHEA)对成骨细胞的选择性作用机制。方法 构建Erβ沉默表达的pLVTHM-GFP/Erβ-shRNA质粒和ERβ3高表达的pWPT-Erβ重组质粒，转染人成骨细胞系hMG63，使hMG63细胞中的Erβ沉默表达（命名为hMG63-shERβ细胞）和高表达（命名为hMG63-Erβ细胞），分别给予DHEA(1 × 10-7 mol/L)、加或不加入U0126、依托泊苷作用后，流式细胞仪检测细胞的增殖和凋亡情况；同时利用逆转录PCR技术检测DHEA对hMG,63细胞雌激素受体亚型mRNA表达的影响。结果hMG63-Erβ和hMG63-shERβ细胞中Erβ基因的相对表达量分别比hMG63细胞升高7.39倍和降低为17％。DHEA对hMG63-shERβ、hMG63-Erβ和对照细胞hMG63细胞Erβ mRNA的表达有升调节作用，而对Erα mRNA的表达无明显影响。与hMG63-shERβ细胞相比，DHEA能明显促进Erβ高表达时（即hMG63-Erβ细胞）和对照细胞hMG63的细胞增殖[增殖指数分别为(81.6±7.6)％、(75.0±5.3)％，P均＜0.05]并拮抗细胞的凋亡[凋亡率分别为(12.2±1.6)％、(14.6±1.5)％，P均＜0.01]，该作用可被U0126部分阻断[增殖指数分别为(33.2±2.0)％、(41.2±2.4)％，凋亡率分别为(40.5±4.3)％、(43.3±4.1)％，P均＜0.05]。结论优势表达的Erβ是介导DHEA对成骨细胞选择性作用的主要受体。
目的：明確Erβ介導脫氫錶雄酮(DHEA)對成骨細胞的選擇性作用機製。方法 構建Erβ沉默錶達的pLVTHM-GFP/Erβ-shRNA質粒和ERβ3高錶達的pWPT-Erβ重組質粒，轉染人成骨細胞繫hMG63，使hMG63細胞中的Erβ沉默錶達（命名為hMG63-shERβ細胞）和高錶達（命名為hMG63-Erβ細胞），分彆給予DHEA(1 × 10-7 mol/L)、加或不加入U0126、依託泊苷作用後，流式細胞儀檢測細胞的增殖和凋亡情況；同時利用逆轉錄PCR技術檢測DHEA對hMG,63細胞雌激素受體亞型mRNA錶達的影響。結果hMG63-Erβ和hMG63-shERβ細胞中Erβ基因的相對錶達量分彆比hMG63細胞升高7.39倍和降低為17％。DHEA對hMG63-shERβ、hMG63-Erβ和對照細胞hMG63細胞Erβ mRNA的錶達有升調節作用，而對Erα mRNA的錶達無明顯影響。與hMG63-shERβ細胞相比，DHEA能明顯促進Erβ高錶達時（即hMG63-Erβ細胞）和對照細胞hMG63的細胞增殖[增殖指數分彆為(81.6±7.6)％、(75.0±5.3)％，P均＜0.05]併拮抗細胞的凋亡[凋亡率分彆為(12.2±1.6)％、(14.6±1.5)％，P均＜0.01]，該作用可被U0126部分阻斷[增殖指數分彆為(33.2±2.0)％、(41.2±2.4)％，凋亡率分彆為(40.5±4.3)％、(43.3±4.1)％，P均＜0.05]。結論優勢錶達的Erβ是介導DHEA對成骨細胞選擇性作用的主要受體。
목적：명학Erβ개도탈경표웅동(DHEA)대성골세포적선택성작용궤제。방법 구건Erβ침묵표체적pLVTHM-GFP/Erβ-shRNA질립화ERβ3고표체적pWPT-Erβ중조질립，전염인성골세포계hMG63，사hMG63세포중적Erβ침묵표체（명명위hMG63-shERβ세포）화고표체（명명위hMG63-Erβ세포），분별급여DHEA(1 × 10-7 mol/L)、가혹불가입U0126、의탁박감작용후，류식세포의검측세포적증식화조망정황；동시이용역전록PCR기술검측DHEA대hMG,63세포자격소수체아형mRNA표체적영향。결과hMG63-Erβ화hMG63-shERβ세포중Erβ기인적상대표체량분별비hMG63세포승고7.39배화강저위17％。DHEA대hMG63-shERβ、hMG63-Erβ화대조세포hMG63세포Erβ mRNA적표체유승조절작용，이대Erα mRNA적표체무명현영향。여hMG63-shERβ세포상비，DHEA능명현촉진Erβ고표체시（즉hMG63-Erβ세포）화대조세포hMG63적세포증식[증식지수분별위(81.6±7.6)％、(75.0±5.3)％，P균＜0.05]병길항세포적조망[조망솔분별위(12.2±1.6)％、(14.6±1.5)％，P균＜0.01]，해작용가피U0126부분조단[증식지수분별위(33.2±2.0)％、(41.2±2.4)％，조망솔분별위(40.5±4.3)％、(43.3±4.1)％，P균＜0.05]。결론우세표체적Erβ시개도DHEA대성골세포선택성작용적주요수체。
Objective To investigate the selective mechanism of dehydroepiandrosterone (DHEA)for osteoblast via ERβ. Methods High expression of ERβ in hMG63-ERβ group ( infected with pWPTERβ), gene silencing of ERβ in hMG63-shERβ group (infected with pLVTHM-GFP/ERβ-shRNA) and hMG,63 group (control) were cultured and treated with 1 × 10-7 mol/L DHEA, with or without U0126 and etoposide. The proliferation and apoptosis of hMG63 were evaluated by flow cytometry. The mRNA level of estrogen receptor subtype was detected by reverse transcription-PCR. Results The expression of ERβ in hMG63-ERβ group and hMG63-shERβ group were increased 7. 39 times and decreased 17％ compared with that in hMG63 group (control). DHEA could increase ERβ expression in hMG63 in each group, however, it did not influence the expression of ERα mRNA. When the level of ERβ was high, DHEA could accelerate the proliferation [proliferation index were ( 81.6 ± 7.6) ％ in hMG,63-ERβ, ( 75.0 ± 5.3 ) ％ in hMG63, P ＜ 0. 05]and inhibit the apoptosis [apoptosis rate were ( 12.2 ± 1.6) ％ in hMG63-ERβ, ( 14. 6 ± 1.5 ) ％in hMG63, P ＜0. 0 1], which was blocked by U0126 [proliferation index were (33. 2 ± 2. 0)％ in hMG63-ERβ, (41.2 ± 2. 4) ％ in hMG63, apoptosis rate were (40. 5 ± 4. 3 ) ％ in hMG63-ERβ, (43.3 ± 4. 1 ) ％in hMG63, all P ＜0. 05]. When the expression of ERβ was silenced, DHEA could not inhibit the apoptosis of hMG63 anymore. Conclusion DHEA selectively act on osteoblasts via the dominant expression of ERβ.
