Abstract
Objective: This study was to investigate the influence of homocysteine on collagen and Angiotensin II type 1 receptor expression in primary fibroblast cells in vitro and their relationship with valsarta, an Angiotensin II type 1 receptor antagonist. Design and method: The primary culture of fibroblast cells of aortic adventitia of rats was performed using tissue explant method. Purified cells at passage 3 to 5 were used for the experiment. After serum starvation for 24 hours, the cells were divided into control group (no homocysteine and no valsartan), Hcy groups and Hcy-Val group. The cells in Hcy groups were stimulated by 100 μmol/L L- homocysteine for 12 hours (12-hour-Hcy group), 24 hours (24-hour-Hcy group) and 48 hours (48-hour-Hcy group). The Hcy-Val group was stimulated by 100 μmol/L L-homocysteine for 48 hours together with 100 nmol/L valsartan 30 min ahead of 48 hours. Western blot was performed to detect Type I collagen and Angiotensin II type 1 receptor expression. Results: (1)Type I collagen expression was increased in 12-hour-Hcy group, 24-hour-Hcy group and 48-hour-Hcy group than in control group(P < 0.05). Type I collagen expression in Hcy-Val group was slightly higher than in control group with no statistical significance. However, its expression was significantly decreased in Hcy-Val group than in 48-hour-Hcy group(P < 0.05). (2) Angiotensin II type 1 receptor expression was increased in Hcy groups than in control group, especially in 24-hour-Hcy group and 48-hour-Hcy group(P < 0.05). Meanwhile, Angiotensin II type 1 receptor expression was significantly decreased in Hcy-Val group than in 48-hour-Hcy group(P < 0.01). Conclusions: Our results showed that homocysteine might promote the Type I collagen and Angiotensin II type 1 receptor expression in primary adventitia fibroblast cells, while Angiotensin II type 1 receptor antagonist valsartan treatment could prevent this effect. These indicated that homocysteine might induce collagen accumulation in vascular adverntitia by stimulating renin-angiotensin system and this mechanism might be mediated by Angiotensin II type 1 receptor.
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