Dissertation
Dissertation > Medicine, health > Pharmacy > Pharmacology

5-HT1B/1D-receptor Mechanism of Pulmonary Artery Smooth Muscle Cell Proliferation and Mitogenesis in the Rat

Author GaoShanHong
Tutor WangHuaiLiang
School China Medical University
Course Pharmacology
Keywords Pentateuch tryptamine Sumatriptan SB224289 BRL15572 Pulmonary Smooth muscle cells 5-HT1B/1D receptor Aortic Selective 5-HT1B/1D receptor agonist 5-HT1B receptor antagonist 5-HT1D receptor antagonist Cells Mitosis Vascular structure remodeling Pulmonary hypertension
CLC R96
Type PhD thesis
Year 2003
Downloads 122
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Objective pulmonary vascular structure remodeling of pulmonary hypertension of the basic pathological changes. Pulmonary vascular smooth muscle cell proliferation and hypertrophy of pulmonary vascular structure remodeling the main feature. 5-HT in the pathogenesis of pulmonary hypertension has a very important role. Study found Monocrotaline induced pulmonary hypertension in rats with pulmonary blood flow in the 5-HT levels and pulmonary vascular smooth muscle 5-HT 1B receptor mRNA increases. 5-HT 1B receptor knockout mice were resistant to hypoxia-induced pulmonary hypertension and pulmonary vascular structure remodeling. 5-HT is an important vasoactive substances ,5-HT only stimulated contraction of pulmonary artery smooth muscle cell proliferation and hypertrophy may cause it, and with a variety of cell growth factors in synergy. Many studies have shown that 5-HT's mitogenic effect is achieved by cell surface receptors, but the 5-HT on pulmonary artery smooth muscle cell mitogenic receptor subtype involved in the mechanism has not been able to fully elucidated. Therefore we assume that 5-HT 1B receptors in pulmonary artery smooth muscle cell proliferation and pulmonary vascular structure remodeling plays an important role, and do the following studies: 1. Rat vascular smooth muscle cells in vitro methods, analysis by flow cytometry and 3 HTdR incorporation in cell proliferation was observed indicators of relevance for the study of drugs on cell proliferation and mitosis influence and role mechanisms to establish a reliable experimental methods. 2. Observe the 5-HT and 5-HT 1B/1D receptor agonist sumatriptan right pulmonary artery smooth muscle cells mitogenic effects of selective 5-HT 1B by antagonist SB 224289 and selective 5-HT 1D receptor antagonist BRL 15572 on 5-HT-induced pulmonary artery smooth muscle cell proliferation , and explore the 5-HT on pulmonary artery smooth muscle cell mitogenic effect of 5-HT 1B receptor mechanism. Explore the 5-HT 1B receptors and 5-HT 1D receptors in pulmonary vascular structure remodeling in rats. 3. Observe the 5-HT and 5-HT 1B/1D receptor agonist sumatriptan right pulmonary artery smooth muscle cells and vascular smooth muscle cells mitogenic effects of differences. Compare selective 5-HT 1B receptor antagonists and selective 5-HT 1D receptor antagonist on 5-HT-induced pulmonary vascular smooth muscle cell proliferation and vascular smooth muscle the impact of differences in cell proliferation explore 5-HT aortic smooth muscle cells mitogenic mechanism. According to the aorta and pulmonary artery smooth muscle cells of 5-HT mitogenic response difference and 5-HT 1B receptor antagonists on the aorta and pulmonary artery smooth muscle cell proliferation differences, explore the 5 - HT 1B receptor antagonists become highly selective anti pulmonary hypertension drug lead compounds possibilities. Method 1. Attached method using isolated rat pulmonary artery smooth muscle cells and vascular smooth muscle cells. Containing 20? S in DMEM primary cultured smooth muscle cells, containing 5? S Vehicle in DMEM for smooth muscle cells subcultured. FBS with different concentrations stimulate cell growth, and the flow cytometry analysis' H TdR incorporation method a cell proliferation index of the correlation. (2) applying 5 a HT, sumatriptan and selective 5-HTIB receptor antagonist SB224289, a selective 5 HTID receptor antagonist BRL15572 other processing factors. With four wow salt (M, I'r) colorimetric count smooth muscle cell proliferation was observed 5-HT, sumatriptan, SB224289 and BRL15572 on smooth muscle cell proliferation. 3, with, H a thymus addicted throat doped DNA synthesis was observed, and to compare the experimental group and the control group cPm value of specific 5-HT receptor agonist and 5-HT receptor antagonists on the pulmonary artery and aortic smooth muscle cells and inhibition of DNA synthesis to promote, demonstrate 5-HTI. 5-HT receptor is induced pulmonary artery smooth muscle cell mitosis role in spots. Explore the 5-HT of aortic smooth muscle cells mitogenic mechanism. Research selective 5 HT; B receptor antagonist SB2242s9, selective 5 HT,. Receptor antagonist BRL15572 on 5-HT-induced smooth muscle cell mitosis impact. 4 Flow cytometry analysis of cell cycle, with M, rr detect cell proliferation was observed, and to explore the 5-HT effects on cell division cycle and selective 5-HT receptor antagonists affect smooth muscle cell mitosis role in cellular cycles. Results 1. Cultured rat vascular smooth muscle cell proliferation and cell flow cytometry method investigated indicators (PI and SPF) and, H a TdR incorporation experiments indicator (cPm) there is a very significant correlation. Flow cytometry and live cell staining techniques MTr combined investigation of cell proliferation and mitosis than alone an indicator of cell proliferation and more accurately reflect the situation in mitosis. 2.5 a HT right pulmonary artery smooth muscle cells mitogenic mechanism of action (1) 5-HT and sumatriptan on PASMC the mitogenic effect M bamboo assay 5 a HT10 a 6 moFL a 10 a 'moFL can promote pulmonary artery smooth muscle cells proliferation, proliferation rates were 256% ± 18%; 228% soil 29%; 201% soil 17%. Sumatriptan 10 a 6 moFL a 10 a 8 moFL also promote pulmonary artery smooth muscle cell proliferation, proliferation rates were from 5% ± 2 one percent; 220% ± 25% and 199% ± 25%. 5-HT from 20 a 6 a 10 a 'moFL and sumatriptan from 10 a 6 a 10 a \HT10 a 5 moFL a 10 a 7 moFL cell proliferation index (Pro-liferation Index) is 23.7% Land 1.1% and 22.6% ± 0.7%, 20.5% ± 0.8%; S phase cell fraction (S a Phase Cell Fraetion) of 8.5% Land 1.3%, 6.9% ± 0.4%, 5.9% ± 0.5%. sumatriptan 10 a, 10 a 6 moUL proliferation index was 23.9% ± 0.8% and 21.2% ± 0.8%; S phase cell fraction was 8.8% ± 0.3% and 6.6% ± 0.5% of the control group and S-phase cells proliferation index scores were 17.59% ± 0.5% and 0.7%, 4.68% soil. 5-HT and sumatriptan can promote pulmonary artery smooth muscle cells from the Go / G, into S phase, and then into the GZ / M phase. 5-HT on pulmonary artery smooth muscle cells mitogenic effects may have 5-HT: 2 1. receptor participate. (2) SB2242s9 and BRL15572 on 5-HT-induced PASMC proliferation assay MTr SB22428, from 10 a 'moFL to 10 a' \

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