Influence of different culture conditions on the phenotype,proliferation and cytoskeleton of VSMCs from rats / 解剖学报

ObjectiveTo study the influence of different culture conditions in vitro on phenotype, proliferation and cytoskeletal proteins expression of vascular smooth muscle cells(VSMCs). Methods The cultured VSMCs from rat aorta were divided into six groups: P2 control,P2 starvation,P4 control,P4 starvation,P6 control and P6 starvation. The proliferating cells were labeled by 5-bromodeoxyuridine (5-BrdU); The mRNA expression of smooth muscle 22 alpha (SM22α) was detected by reverse transcription-polymerase chain reaction method (RT-PCR); The cytoskeletal proteins including SMα-actin,β-Tubulin and Desmin were observed through immunohistochemical staining. Results With the increase of cell passage, cytoskeletal proteins expression of VSMCs decreased,cellular organs increased and secretory vesicles were abundant; in serum-free cultured cells mitochondria increased and electron density enhanced in cytoplasm of VSMCs.On the contrary the expression of SMα-actin decreased, and the expression of SMα-actin increased. The expression of β-Tublin and Desmin decreased more obviously, and at 6 passages failed to express. Conclusion The conditioned medium and serum-free had the different effects on the phenotype,proliferation and cytoskeleton of VSMCs in different passage, and there was internal relationship among them. The internal relationship played an important role in the maintaining of cell morphology, contractile function and vascular remodeling. The disappearance of expression of β-Tubulin and desmin might have important biological significance.

Establishing an organic model of SMC proliferation with cultured aorta of rats and exploring the underlying mechanism / 生物医学工程学杂志

To study the mechanism of proliferous vascular disease as well as its prevention and treatment, an organic model was established with cultured aortas of rats, and the mechanism there-in invloved was probed. Immunostaining histology showed that smooth muscle cell (SMC) proliferation was observed in the aorta segments of rats, after their endothelia being injured and cultured in vitro with 20% fetal bovine serum. After being cultured for 5 days, various degrees of proliferation of SMC on cultured artery segments were observed by HE staining, and conspicuous plaques were developed after being cultured for 13 days. The proliferous SMC was also observed by Brdu labeling. RT-PCR examination showed that the mRNA expression of hypertension-related gene-1 (Hrg-1) and smooth muscle 22 alpha (SM22a) in the aortas decreased with the prolongation of culture time, and completely disappeared after being cultured for 13 days . But when cultured in vitro for ten days, the ET-1 content of supernatant and the proliferous SMC labeled by Brdu increased obviously and the expressions of Hrg-1 and SM22a decreased after the endothelium was destroyed. Compared with the injured endothelium groups, the proliferous SMC of injured endothelium plus BQ123 groups decreased visibly. The same significant differences between serum groups and serum-free groups were also observed. These results suggest that the culturing of rat aorta segments in vitro can induce the proliferation of SMC and the transform of phenotype from contractile type to synthetic type. The ET-1 and serum are the main factors in the proliferation of SMC and in the transform of phenotype. This organic model could serve as a good experimental platform for the researches into the mechanism of proliferous vascular disease as well as its prevention and treatment.

Study of aspirin on inhibiting the atherosclerotic plaque rupture and MMP-2 expression of abdominal aorta in atherosclerotic rabbits / 中国药理学通报

Aim To study the effects of aspirin on increasing the atherosclerotic plaque stability and its possible mechanisms.Methods The hyperlipidemic atherosclerotic model was generated in male New Zealand rabbits given high fat diet and endothelial abrasion of abdominal aorta.These rabbits were then treated with aspirin 5～20 mg?kg-1 for 4 weeks.At experimental end,the plaques were evoked into rupture by injection of Russell's viper venom and histamine.Areas of thrombosis on atherosclerotic aorta were determined by image analysis,morphologic character of plaque rupture was examined by light microscope,the protein expression of macrophages was detected by immunohistochemistry,and the mRNA expression of COX-2 and MMP-2 was determined by hybridization in situ,respectively.Results Aspirin at doses of 5～10 mg?kg-1 was able to inhibit thrombosis on atherosclerotic plaque(P

INFLUENCE OF CALCITONIN GENE-RELATED PEPTIDE ON THE PROLIFERATION AND PHENOTYPE TRANSFORMATION OF VASCULAR SMOOTH MUSCLE CELLS / 解剖学报

Objective To investigate whether calcitonin gene-related peptide(CGRP) can influence the proliferation and phenotypic modulation of vascular smooth muscle cells(VSMCs),and what is the relationship between them. Methods Vascular smooth muscle cells(VSMCs) were cultured respectively with rat aorta cultivated for 8 days in vitro and with normal aorta(not culture) through the explant-attached method,and CGRP was added into the culture medium of the experimental groups.The proliferation of cells was labeled by 5-bromodeoxyuridine(5-BrdU) with immunocytochemical method,and the mRNA expression of hypertension-related gene-1(HRG-1) and smooth muscle 22 alpha(SM22?) were determined by RT-PCR. Results The proliferating cells labeled by BrDU from the aorta cultured for 8 days in vitro were increased notablly and the mRNA expression of HRG-1 and SM22? were decreased.While the VSMCs were cultured in the culture medium containing CGRP,the proliferous cells labeled by BrdU were obviously decreased and the mRNA expression of HRG-1 and SM22? were significantly increased.Conclusion It is showed that CGRP could inhibit the proliferation of VSMCs and change the phenotype of VSMCs from synthesize to contractile type.It might be a good cellular model which provides a good experimental platform to research the proliferating vascular disease as well as its prevention and treatment.

THE CHANGES OF THE TOTAL CHOLESTEROL,HIGH DENSITY LIPOPROTEIN, OXIDIZED LOW DENSITY LIPOPROTEIN, NITRIC MONOXIDE AND LIPOPEROXIDE IN SERUM AFTER THE ARCUATE NUCLEUS LESIONED / 解剖学报

Objective To study the influence of the hypothalamic arcuate nucleus on the development of atherosclerosis. Methods The changes of the total cholesterol, high density lipoprotein, oxidized low density lipoprotein, nitric monoxide and lipoperoxide in serum were investigated after the arcuate nucleus lesioned with monosodium glutamate. Results The rat with arcuate nucleus lesioned had significant higher levels of total cholesterol, oxidized low density lipoprotein and lipoperoxide in serum than that of control subjects, but the level of nitric monoxide was lower and the high density lipoprotein in serum had no change.Conclusion This study indicated that hypothalamic arcuate nucleus might exert regulative effect in the development of atherosclerosis.