A cell culture model using rat coronary artery adventitial fibroblasts to measure collagen production.

BACKGROUND: We have developed a rat cell model for studying collagen type I production in coronary artery adventitial fibroblasts. Increased deposition of adventitial collagen type I leads to stiffening of the blood vessel, increased blood pressure, arteriosclerosis and coronary heart disease. Although the source and mechanism of collagen deposition is yet unknown, the adventitia appears to play a significant role. To demonstrate the application of our cell model, cultured adventitial fibroblasts were treated with sex hormones and the effect on collagen production measured. METHODS: Hearts (10-12 weeks) were harvested and the left anterior descending coronary artery (LAD) was isolated and removed. Tissue explants were cultured and cells (passages 2-4) were confirmed as fibroblasts using immunohistochemistry. Optimal conditions were determined for cell tissue harvest, timing, proliferation and culture conditions. Fibroblasts were exposed to 10-7 M testosterone or 10-7 M estrogen for 24 hours and either immunostained for collagen type I or subjected to ELISA. RESULTS: Results showed increased collagen staining in fibroblasts treated with testosterone compared to control and decreased staining with estrogen. ELISA results showed that testosterone increased collagen I by 20% whereas estrogen decreased collagen I by 15%. CONCLUSION: Data demonstrates the usefulness of our cell model in studying the specific role of the adventitia apart from other blood vessel tissue in rat coronary arteries. Results suggest opposite effects of testosterone and estrogen on collagen synthesis in the rat coronary artery adventitial fibroblasts.

Social stress increases blood pressure and cardiovascular pathology in a normotensive rat model.

Territorial stress (TS) elevates blood pressure (BP) in several mammalian species. However, cardiovascular pathology following chronic stress has not been consistently shown in a non-genetic hypertension model. Therefore, the hypothesis tested was that social stress would directly increase: BP, collagen deposition in coronary and mesenteric arteries, and myocardial fibrosis. Wistar-Kyoto (WKY) male rats, four weeks of age, were divided into one of three groups: controls (n = 9), territorial stress (TS, n = 12), and social isolation followed by territorial stress (SITS, n = 11). Blood pressure was measured biweekly, and blood samples biweekly for serum testosterone, corticosterone, epinephrine and norepinephrine. Blood pressure significantly increased in the TS (130 mmHg, p < 0.05) and SITS (150 mmHg, p < 0.05) groups, compared to controls (120 mmHg, ANOVA, F = 6.7, p < 0.001). Coronary collagen was increased 47% in the TS group and 90% in the SITS group compared to controls (p < 0.05). The coronary wall/lumen ratio increased significantly (45%, p < 0.05) in the SITS group compared to the controls. Myocardial fibrosis was increased 27% in the TS group and 74% in the SITS group compared to controls (p < 0.05). In conclusion, stress treatments increased BP and cardiac pathology in a normotensive rat strain.