Sex differences in the lung ACE/ACE2 balance in hypertensive rats

Figures

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  Figure 1. Lung ACE and ACE2 mRNA and protein expression in male and female normotensive (Wistar) and hypertensive (SHR) rats The gene expression of (A) ACE and (B) ACE2 was quantitatively measured by RT-PCR and normalized to cyclophilin and is expressed as the % of male Wistar rats. (C) Lung ACE/ACE2 gene expression ratio was expressed as 2−ΔCt/2−ΔCt. The protein expression of (D) ACE and (E) ACE2 was determined by immunoblotting of lung membrane proteins and normalized to β-actin and is expressed as the % of male Wistar rats (Supplementary Figures S3 and S4). The values represent individual measurements and the means ± SEMs. *P<0.05; **P<0.01; ***P<0.001.

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  Figure 2. Lung activity of ACE and ACE2 and lung concentration of their respective products, Ang II and Ang-(1-7), in male and female normotensive (Wistar) and hypertensive (SHR) rats The activities of (A) ACE and (B) ACE2 were assayed by a fluorogenic method. (C) Lung ACE/ACE2 activity ratio. Lung (D) Ang II and (E) Ang-(1-7) concentrations were determined by ELISA. (F) Lung Ang II/Ang-(1-7) peptide concentration ratio. The values represent individual measurements and the means ± SEMs. *P<0.05; **P<0.01; ***P<0.001.

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  Figure 3. Serum ACE and ACE2 concentrations and their correlation with blood pressure in male and female normotensive (Wistar) and hypertensive (SHR) rats Serum (A) ACE and (B) ACE2 concentrations were determined by ELISA. The values represent individual measurements and the means ± SEMs. **P<0.01 and ***P<0.001. Correlations between the serum (C) ACE and (D) ACE2 concentrations and tail-cuff blood pressure. The correlation coefficients (r) and P-values were obtained by Pearson’s correlation test, and lines were obtained by linear regression plotting.

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  Figure 4. Lung ADAM17 activity and its correlation with lung ACE2 protein and serum abundance in male and female normotensive (Wistar) and hypertensive (SHR) rats (A) Lung ADAM17 activity was determined by a fluorescent enzymatic assay in the presence or absence of TAPI 0. The TAPI 0 sensitive enzymatic activity was expressed as RFUs/µg/h. The values represent individual measurements and the means ± SEMs. The absence of a correlation between lung ADAM17 activity and (B) lung ACE2 protein or (C) serum ACE2 concentration. The correlation coefficients (r) and P-values were obtained by Pearson’s correlation test, and lines were obtained by linear regression plotting.

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  Figure 5. The relationship between the lung expression and serum concentrations of ACE and ACE2 (A) Correlation between serum ACE concentration and lung ACE mRNA level. (B) The absence of a correlation between serum ACE concentration and lung ACE protein abundance. (C) The absence of a correlation between lung ACE protein abundance and lung ACE mRNA level. (D) The absence of a correlation between serum ACE2 concentration and lung ACE2 mRNA level. (E) The absence of a correlation between serum ACE2 abundance and lung ACE2 protein abundance. (F) Correlation between lung ACE2 protein abundance and lung ACE2 mRNA level. The correlation coefficients (r) and P-values were obtained by Pearson’s correlation test, and lines were obtained by linear regression plotting.

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  Figure 6. Lung ACE and ACE2 mRNA expression in normotensive and hypertensive patients Normalized (A) ACE mRNA and (B) ACE2 mRNA expression data and the (C) ACE to ACE2 mRNA ratio from normotensive and hypertensive patients whose lung samples were available in the GTEx database. Boxplots show the minimum, first quartile, median, third quartile, and maximum values. Each dot represents an individual data point.