The mechanism of matrix-synthesis and-degradation in vascular smooth muscle cells.

血管平滑肌细胞基质合成和降解的机制。

• 批准号: 10670220
• 负责人: KATO Seiya
• 金额: $ 1.92万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670220/
• 关键词: Atherosclerosis; Smooth muscle, vascular; Phenotype; Matrix metalloproteinase; Cell cycle; p21 Waf-1; Adenovirus vector; マトリッックスメタロプロテイナーゼ; アデノウイルスベクター

To understanding the relationship between phenotypic modulation of vascular smooth muscle cells (SMC), and matrix remodeling process, in vitro culture model for the phenotypic modulation of SMC was established. The culture conditions were verified by the alteration of serum-concentration and final cell densities. The expressions of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) were determined with Western blotting and enzyme immunoassays. Collogenolytic activity and gelationolytic activity were also measured. The Expressions and activities of MMP-1, TIMP-1-3, and MMP-1rrIMP-1 expressions were maximal in the cells displaying the proliferative phenotype. MMP-2 expression was not changed with SMC phenotypes. The adenovirus vector expressing TIMP-1 was prepared and the analysis for SMC phenotypes or functions using this vector is addressing.The amount of extra-cellular matrix also regulates SMC phenotypes via the function of cell-cycle regulatory genes. The expression patte … More rn of p2lWaf-1, cyclin-dependent kinase inhibitor was observed in either cultured SMC or human atherosclrotic lesions. p2lWaf-1 expression was enhanced in the proliferating cells in accordance with G0-G1 transition. In vivo, the expression was dominantly detected in neo-intimal atherosclerotic lesions. p21Waf-1 may contribute not only in cell-cycle arrest, but also in an appropriate cell cycle progression. Adenovirus vector expressing p21Waf-1 induced cell cycle arrest and hypertrophy in SMC, but not promoted re-differentiation and apoptosis.In conclusion, Matrix-degradation is an important function of SMC, which is coordinately regulated with SMC phenotypes. These SMC functions may be mediated by both endogenous and exogenous growth factor signalings. Proliferation is considered to be tightly regulated by cell-cycle regulatory proteins, such as p21Waf-1, of which function may modulate SMC phenotypes. These finding may contribute to understanding the biology of vascular smooth muscle cells. The adenovirus-mediated gene transfer method used in this project, may imply clinical alteration of human atherosclerosis in the therapeutic implications. Less

为了解血管平滑肌细胞（SMC）表型调控与基质重塑过程的关系，建立了SMC表型调控的体外培养模型。通过血清浓度和最终细胞密度的变化来验证培养条件。采用免疫印迹法和酶免疫分析法检测基质金属蛋白酶（MMPs）及其抑制剂（TIMPs）的表达。还测量了胶原分解活性和胶凝分解活性。MMP-1、TIMP-1-3和MMP-1 rrIMP-1的表达和活性在显示增殖表型的细胞中最高。MMP-2表达不随SMC表型改变而改变。制备了表达TIMP-1的腺病毒载体，并利用该载体对SMC的表型和功能进行了分析，细胞外基质的量也通过细胞周期调控基因的功能调节SMC的表型。表达式patte ...更多信息 在培养的SMC或人动脉粥样硬化病变中观察到细胞周期蛋白依赖性激酶抑制剂p21 Waf-1的rn。p21 Waf-1在增殖期细胞中表达增强，与G 0-G1期相一致。在体内，主要在新生内膜动脉粥样硬化病变中检测到该表达。p21 Waf-1不仅参与细胞周期阻滞，而且参与细胞周期的适当进程。腺病毒介导的p21 Waf-1表达可诱导SMC细胞周期阻滞和肥大，但不促进再分化和凋亡。这些SMC功能可能是由内源性和外源性生长因子信号传导介导的。增殖被认为是由细胞周期调控蛋白，如p21 Waf-1，其功能可能会调节SMC表型的严格调节。这些发现可能有助于了解血管平滑肌细胞的生物学。本研究所采用的腺病毒介导的基因转移方法，可能对人类动脉粥样硬化的临床改变具有治疗意义。少

项目成果

Seiya Kato, Miki Yamaguchi, Teruhiko Fujii, Naohisa Miyagi, Mizuhiko Terasaki, Tetsuya Hamada, Yasuo Sugita, Minoru Morimatsu: "Over-expression of p21Waf-1 in vascular smooth muscle cells : Regulation oinproliferation, differentiation, and cell size."Exp

Seiya Kato、Miki Yamaguchi、Teruhiko Fujii、Naohisa Miyagi、Mizuhiko Terasaki、Tetsuya Hamada、Yasuo Sugita、Minoru Morimatsu：“血管平滑肌细胞中 p21Waf-1 的过度表达：增殖、分化和细胞大小的调节。”Exp

Seiya Kato他: "Basic fibroblast growth factor regulatea extracellular matrix and contractile protein expression independent of proliferation in vascular smooth muscle cells"In Vitro Cell Dev Biol-Animal. 34. 341-346 (1998)

Seiya Kato 等人：“碱性成纤维细胞生长因子独立于血管平滑肌细胞的增殖调节细胞外基质和收缩蛋白表达”In Vitro Cell Dev Biol-Animal 34. 341-346 (1998)。

Seiya Kato他: "Over-expression of p21 Waf-1 in vascular smooth muscle cells : Regulation in proliferation,differentiation,and cell size." Exp Mol Pathol. (in print).

Seiya Kato 等人：“血管平滑肌细胞中 p21 Waf-1 的过度表达：增殖、分化和细胞大小的调节”（Exp Mol Pathol）。

Seiya Kato, Hideo Yasukawa, Teruhiko Fujii, Miki Ymaguchi, Naohise, Miyagi, Kenichi Okamoto, Yoshihiro Wada, Morimatsu, Jonathan. C. Fox.: "Coordinate regulation of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 expression in human

加藤圣哉、安川秀夫、藤井辉彦、山口美纪、尚濑、宫城、冈本健一、和田义弘、森松、乔纳森。

Seiya Kato他: "Corrdinate regulation of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 expression in vascular smmothe muscle cells."Connect Tissue Res. in print.

Seiya Kato 等人：“血管平滑肌细胞中基质金属蛋白酶-1 和金属蛋白酶-1 组织抑制剂表达的协调调节。”《Connect Tissue Res》印刷版。