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(Hypertension. 1997;30:880-885.)
© 1997 American Heart Association, Inc.

Articles

NHE-1 Protein in Vascular Smooth Muscle and Lymphocytes From the Spontaneously Hypertensive Rat

Michael S. LaPointe; Minghao Ye; Robert Bacallao; ; Daniel Batlle

From the Northwestern University Medical School and Chicago Veterans Affairs Health Care Authority, Lakeside Division, Chicago, Ill.

Correspondence to Daniel Batlle, MD, Northwestern University Medical School, Department of Medicine/Division of Nephrology, Searle 10-475, 303 E Chicago Ave, Chicago, IL 60611.

Abstract The present study examined the abundance of NHE-1 protein in cultured vascular smooth muscle cells (VSMCs), freshly isolated thymocytes, and fresh aortic tissue from spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) rats. Two sets of affinity-purified antibodies (Ab_(765-778) and Ab_(698-711)) against different epitopes of the NHE-1 isoform of the Na⁺-H⁺ antiporter were used. Each set of antibodies recognized a major protein band at 105 to 110 kD that was more abundant in protein lysates prepared from cultured VSMCs from the SHR than those from WKY rats (Ab_(765-778) 0.047±0.011 vs 0.010±0.002 O.D. units/10 µg protein, P<.001 for SHR and WKY, respectively; and Ab_(698-711) 0.173±0.026 vs 0.087±0.028 O.D. units/10 µg protein, P<.05, for SHR and WKY, respectively). The increase in NHE-1 protein abundance in cultured VSMCs from the SHR was associated with a greater V_max of the Na⁺-H⁺ antiporter as compared to those from WKY rats (17.93±2.07 vs 8.16±1.05 mmol H⁺/min, P<.001, respectively). In contrast to cultured VSMCs, there was no difference in the relative abundance of NHE-1 protein in fresh aortic tissue (0.075±0.018 vs 0.083±0.017 O.D. units/10 µg protein, from SHR and WKY, respectively) or in freshly isolated thymocytes (0.158±0.046 vs 0.226±0.054 O.D. units/10 µg protein, from SHR and WKY, respectively). We conclude that the increase in the V_max of the Na⁺-H⁺ antiporter in cultured VSMCs from the SHR, compared to those from WKY rats, is due, at least in part, to increased levels of NHE-1 protein.

Key Words: sodium-hydrogen antiporter • NHE-1 • rats, inbred SHR • muscle, smooth • Western blot

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