Endothelial dependent relaxation was recently reported to be decreased in SHR, which suggests the dysfunction of endothelial cells. To clarify the functional alteration of endothelial cells, we cultured endothelial cells from rat thoracic aorta, and analyzed the difference of cellular structure and receptor signaling in the presence of bradykinin (BK) between SHR and normotensive Wistar Kyoto rats (WKY). Intracellular free Ca
2+ ([Ca
2+] i), IP
3 and PG-I
2 were measured using an image analyzer (Argus-100 CA, Hamamatsu photonics) with fura-2/AM, radioligand
3H-IP
3 and
125I-6-keto PG-F
1α, respectively.There was no difference either in cellular structure and in the BK-receptor, (Bmax = 266±18 vs 254±20 fmol/10
6cells, Kd = 0.39±0.02 vs 0.04±0.03nM) between SHR and WKY, respectively. However, 10
-9M to 10
-7M BK increased [Ca
2+] i by 2.5 to 5.6 fold in SHR compared with [Ca
2+] i in WKY (p<0.05 n = 8). Since EGTA failed to give higher [Ca
2+] i in SHR, elevation of [Ca
2+] i appeared to be brought about by Ca
2+ influx via the cytoplasmic membrane in SHR rather than intracellular Ca
2+ release from internal stores. SHR also showed higher elevation in IP
3 by 1.2 to 1.6 fold compared with those of WKY at a concentration of 10
-9 M to 10
-7 M BK (p<0.05 vs WKY). This indicates that the activity of PL-C was enhanced in SHR compared with that of WKY. By contrast, the release of. PG-I
2 from the endothelial cells was diminished in SHR by 0.4folds compared with that of WKY (p<0.05 vs WKY). Thus, despite the lack of a difference in cellular structure and BK receptor kinetics between SHR and WKY, cultured endothelial cells from SHR showed a stronger response in [Ca
2+] i and IP
3, and diminished response in PG-I
2 in the presence of BK than the endothelial cells from WKY. These findings indicate that endothelial cells from SHR are biochemically different in culture from those of WKY. Such cellular alteration may play a role in endothelial-related vascular physiology in hypertension.
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