Lipopolysaccharide inhibits proliferation of the cultured vascular smooth muscle cells by stimulating inducible nitric oxide synthase and subsequent activation of guanylate cyclase.
- Author:
Hyoung Chul CHOI
1
;
Sang Gon LEE
;
Jong Ho KIM
;
Joo Young KIM
;
Uy Dong SOHN
;
Jeoung Hee HA
;
Kwang Youn LEE
;
Won Joon KIM
Author Information
1. Department of Pharmacology, College of Medicine, Yeungnam University, Daegu, South Korea. youny@medical.yeung-nam.ac.kr
- Publication Type:Original Article
- MeSH:
Animals;
Aorta;
Arginine;
Blotting, Western;
Cell Count;
Cell Death;
Gene Expression;
Guanylate Cyclase*;
Immunohistochemistry;
L-Lactate Dehydrogenase;
Muscle, Smooth, Vascular*;
Myosins;
Nitric Oxide;
Nitric Oxide Synthase Type II*;
Nitroprusside;
Rats;
RNA, Messenger
- From:The Korean Journal of Physiology and Pharmacology
2001;5(4):343-351
- CountryRepublic of Korea
- Language:English
-
Abstract:
This study was undertaken to investigate the mechanism of lipopolysaccharide (LPS) and nitric oxide (NO) as a regulator of vascular smooth muscle cell (VSMC) proliferation. VSMC was primarily cultured from rat aorta and confirmed by the immunocytochemistry with anti-smooth muscle myosin antibody. The number of viable VSMCs were counted, and lactate dehydrogenase (LDH) activity was measured to assess the degree of cell death. Concentrations of nitrite in the culture medium were measured as an indicator of NO production. LPS was introduced into the medium to induce the inducible nitric oxide synthase (iNOS) in VSMC, and Western blot for iNOS protein and RT-PCR for iNOS mRNA were performed to confirm the presence of iNOS. Inhibitors of iNOS and soluble guanylate cyclase (sGC), sodium nitroprusside (SNP) and L-arginine were employed to observe the action of LPS on the iNOS-NO-cGMP signalling pathway. LPS and SNP decreased number of VSMCs and increased the nitrite concentration in the culture medium, but there was no significant change in LDH activity. A cell permeable cGMP derivative, 8-Bromo-cGMP, decreased the number of VSMCs with no significant change in LDH activity. L-arginine, an NO substrate, alone tended to reduce cell count without affecting nitrite concentration or LDH level. Aminoguanidine, an iNOS specific inhibitor, inhibited LPS-induced reduction of cell numbers and reduced the nitrite concentration in the culture medium. LY 83583, a guanylate cyclase inhibitor, suppressed the inhibitory actions of LPS and SNP on VSMC proliferation. LPS increased amounts of iNOS protein and iNOS mRNA in a concentration-dependent manner. These results suggest that LPS inhibits the VSMC proliferation via production of NO by inducing iNOS gene expression. The cGMP which is produced by subsequent activation of guanylate cyclase would be a major mediator in the inhibitory action of iNOS-NO signalling on VSMC proliferation.
