Contractile properties of the cultured vascular smooth muscle cells - The crucial role played by RhoA in the regulation of contractility

被引:44
作者
Bi, D [1 ]
Nishimura, J [1 ]
Niiro, N [1 ]
Hirano, K [1 ]
Kanaide, H [1 ]
机构
[1] Kyushu Univ, Grad Sch Med Sci, Angiocardiol Res Inst, Div Mol Cardiol, Fukuoka 812, Japan
关键词
vascular smooth muscle; contraction; RhoA; CPI-17; RNA interference; vascular disease;
D O I
10.1161/01.RES.0000163018.66460.85
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Vascular smooth muscle cells (VSMCs) have a remarkable degree of plasticity and in response to vascular injury, they can change to a dedifferentiated state that can be typically seen in cell cultures. Recently, Y27632, a Rho kinase inhibitor, has been reported to preferentially correct hypertension in a hypertensive rat model. We thus tested the hypothesis that the contraction of the cultured VSMCs might be more dependent on the function of RhoA than the VSMCs in fresh tissue. For this purpose, a tissue-like ring preparation was made using the cultured porcine coronary artery SMCs (CASMCs) and collagen gel ( reconstituted ring: R-ring). The R-ring developed an isometric tension on stimulation by high external K+ or various receptor agonists. The phorbol ester ( a protein kinase C (PKC) activator)- induced contraction of the intact R-ring was greatly inhibited, while the GTP gamma S ( an activator of RhoA)- induced and Ca2+-independent contraction of permeabilized R-ring was greatly enhanced, in comparison to the fresh coronary artery ring. An immunoblot analysis showed the expression levels of RhoA and myosin phosphatase subunits (MYPT1 and PP1c delta) to be up-regulated, while the levels of CPI-17 ( PKC-potentiated protein phosphatase-1 inhibitory protein), h1-calponin and PKC isoforms were downregulated in cultured CASMCs. The knock down of RhoA by RNA interference decreased the contractility of the cultured CASMCs. It is concluded that the contractility of the cultured VSMCs thus appears to be much more dependent on the function of RhoA than VSMCs in fresh tissue. The expression level of RhoA thus plays a crucial role in regulating the contractility of cultured VSMCs.
引用
收藏
页码:890 / 897
页数:8
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