中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2014年
3期
238-244
,共7页
田利民%倪菁%刘静%张陆燕
田利民%倪菁%劉靜%張陸燕
전이민%예정%류정%장륙연
促甲状腺激素%血管内皮细胞%血管平滑肌细胞%动脉粥样硬化
促甲狀腺激素%血管內皮細胞%血管平滑肌細胞%動脈粥樣硬化
촉갑상선격소%혈관내피세포%혈관평활기세포%동맥죽양경화
Thyroid stimulating hormone%Vascular endothelial cells%Vascular smooth muscle cells%Atherosclerosis
目的 通过促甲状腺激素(TSH)对人血管内皮细胞和平滑肌细胞影响的研究,探讨TSH在动脉粥样硬化发生发展过程中的作用.方法 体外培养人血管内皮细胞和平滑肌细胞,采用MTT法检测TSH对细胞活性的影响,实时荧光定量PCR观察TSH对内皮细胞内皮型一氧化氮合酶(eNOS)、前列环素(PGI2)、内皮素1、纤溶酶原激活物抑制剂1(PAI-1)mRNA水平以及平滑肌细胞表型转换的影响;流式细胞仪检测TSH对平滑肌细胞周期的影响,应用免疫组化、Western印迹检测TSH对平滑肌细胞周期素A、D1以及内皮细胞相关因子的蛋白表达.结果 与对照组比较不同浓度TSH作用于内皮细胞后eNOS、PGI2 mRNA水平降低,内皮素1、PAI-1 mRNA水平增加(P＜0.05),eNOS蛋白表达水平逐渐降低,而内皮素1蛋白表达水平逐渐增加(P＜0.05).不同浓度TSH作用于平滑肌细胞后,可以促进平滑肌细胞发生表型转换、促进细胞周期向G2/M期转换,并使细胞周期素D1、A的表达量增加.结论 TSH可能损伤血管内皮细胞功能,促进血管平滑肌细胞增殖.
目的 通過促甲狀腺激素(TSH)對人血管內皮細胞和平滑肌細胞影響的研究,探討TSH在動脈粥樣硬化髮生髮展過程中的作用.方法 體外培養人血管內皮細胞和平滑肌細胞,採用MTT法檢測TSH對細胞活性的影響,實時熒光定量PCR觀察TSH對內皮細胞內皮型一氧化氮閤酶(eNOS)、前列環素(PGI2)、內皮素1、纖溶酶原激活物抑製劑1(PAI-1)mRNA水平以及平滑肌細胞錶型轉換的影響;流式細胞儀檢測TSH對平滑肌細胞週期的影響,應用免疫組化、Western印跡檢測TSH對平滑肌細胞週期素A、D1以及內皮細胞相關因子的蛋白錶達.結果 與對照組比較不同濃度TSH作用于內皮細胞後eNOS、PGI2 mRNA水平降低,內皮素1、PAI-1 mRNA水平增加(P＜0.05),eNOS蛋白錶達水平逐漸降低,而內皮素1蛋白錶達水平逐漸增加(P＜0.05).不同濃度TSH作用于平滑肌細胞後,可以促進平滑肌細胞髮生錶型轉換、促進細胞週期嚮G2/M期轉換,併使細胞週期素D1、A的錶達量增加.結論 TSH可能損傷血管內皮細胞功能,促進血管平滑肌細胞增殖.
목적 통과촉갑상선격소(TSH)대인혈관내피세포화평활기세포영향적연구,탐토TSH재동맥죽양경화발생발전과정중적작용.방법 체외배양인혈관내피세포화평활기세포,채용MTT법검측TSH대세포활성적영향,실시형광정량PCR관찰TSH대내피세포내피형일양화담합매(eNOS)、전렬배소(PGI2)、내피소1、섬용매원격활물억제제1(PAI-1)mRNA수평이급평활기세포표형전환적영향;류식세포의검측TSH대평활기세포주기적영향,응용면역조화、Western인적검측TSH대평활기세포주기소A、D1이급내피세포상관인자적단백표체.결과 여대조조비교불동농도TSH작용우내피세포후eNOS、PGI2 mRNA수평강저,내피소1、PAI-1 mRNA수평증가(P＜0.05),eNOS단백표체수평축점강저,이내피소1단백표체수평축점증가(P＜0.05).불동농도TSH작용우평활기세포후,가이촉진평활기세포발생표형전환、촉진세포주기향G2/M기전환,병사세포주기소D1、A적표체량증가.결론 TSH가능손상혈관내피세포공능,촉진혈관평활기세포증식.
Objective To study the effect of thyroid-stimulating hormone (TSH) on human vascular endothelial cells and smooth muscle cells and to explore the roles of TSH in the development of atherosclerosis.Methods Human vascular endothelial cells and smooth muscle cells were cultured in vitro.MTT method was used to assay the effect of TSH on cell viability.Real-time PCR was used to determine the levels of endothelial nitric oxide synthase (eNOS),prostacyclin(PGI2),endothelin-1 (ET-1),plasminagen activator inhibitor(PAI-1) (mRNA) in endothelial cells and the phenotype transition of smooth muscle cells.The effect of TSH to the cycle of smooth muscle cells was detected by using flow cytometry.Immunocytochemistry and Western blot were used to investigate the expression of the cell cyclin A,D1,and the expression of endothelial cell associated factors eNOS and ET-1.Results Compared with the control group,eNOS and PGI2 mRNA levels decreased while ET-1 and PAI-1 mRNA levels increased when different concentrations of TSH were applied to endothelial cells(P＜0.05).The level of eNOS protein was decreased gradually while the level of ET-1 protein was gradually increased(P＜0.05).Different concentrations of TSH applied to smooth muscle cells could promote the transition of cell cycle phase G2 to phase M and increase the expression of cell cyclin A and D1.Conclusion TSH may damage the function of vascular endothelial cells and promote the proliferation of smooth muscle cells.
