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Reducing TRPC1 Expression through Liposome-Mediated siRNA Delivery Markedly Attenuates Hypoxia-Induced Pulmonary Arterial Hypertension in a Murine Model.

Sun CK, Zhen YY, Lu HI, Sung PH, Chang LT, Tsai TH, Sheu JJ, Chen YL, Chua S, Chang HW, Chen YL, Lee FY, Yip HK - Stem Cells Int (2014)

Bottom Line: By day 28, right ventricular systolic pressure (RVSP), number of muscularized arteries, right ventricle (RV), and lung weights were increased in group 2 than in group 1 and reduced in group 3 compared with group 2.The mRNA expressions of proapoptotic and hypertrophy biomarkers displayed a similar pattern, whereas sarcomere length showed an opposite pattern compared to that of RVSP in all groups.Lipofectamine siRNA delivery effectively reduced TRPC1 expression, thereby attenuating PAH-associated RV and pulmonary arteriolar remodeling.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency Medicine, E-Da Hospital, I-Shou University College of Medicine, Kaohsiung 82445, Taiwan.

ABSTRACT
We tested the hypothesis that Lipofectamine siRNA delivery to deplete transient receptor potential cation channel (TRPC) 1 protein expression can suppress hypoxia-induced pulmonary arterial hypertension (PAH) in mice. Adult male C57BL/6 mice were equally divided into group 1 (normal controls), group 2 (hypoxia), and group 3 (hypoxia + siRNA TRPC1). By day 28, right ventricular systolic pressure (RVSP), number of muscularized arteries, right ventricle (RV), and lung weights were increased in group 2 than in group 1 and reduced in group 3 compared with group 2. Pulmonary crowded score showed similar pattern, whereas number of alveolar sacs exhibited an opposite pattern compared to that of RVSP in all groups. Protein expressions of TRPCs, HIF-1α, Ku-70, apoptosis, and fibrosis and pulmonary mRNA expressions of inflammatory markers were similar pattern, whereas protein expressions of antifibrosis and VEGF were opposite to the pattern of RVSP. Cellular markers of pulmonary DNA damage, repair, and smooth muscle proliferation exhibited a pattern similar to that of RVSP. The mRNA expressions of proapoptotic and hypertrophy biomarkers displayed a similar pattern, whereas sarcomere length showed an opposite pattern compared to that of RVSP in all groups. Lipofectamine siRNA delivery effectively reduced TRPC1 expression, thereby attenuating PAH-associated RV and pulmonary arteriolar remodeling.

No MeSH data available.


Related in: MedlinePlus

Immunofluorescent (IF) staining for γ-H2AX+ cells in medial layer of pulmonary arterioles (PA) and lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) (n = 10). ((a) to (c)) Identification of γ-H2AX+ cells in PA (white arrow) using IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (d) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. ((e) to (g)) Identification of γ-H2AX+ cells in lung parenchyma (white arrow) with IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (h) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. The scale bars in right lower corners represent 20 μm. Blue fluorescence indicates DAPI-stained nuclei. Statistical analysis in (d) and (h) using one-way ANOVA, followed by Bonferroni multiple comparison post hoc test. Symbols (∗, †, ‡) indicate significance (at 0.05 level).
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fig3: Immunofluorescent (IF) staining for γ-H2AX+ cells in medial layer of pulmonary arterioles (PA) and lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) (n = 10). ((a) to (c)) Identification of γ-H2AX+ cells in PA (white arrow) using IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (d) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. ((e) to (g)) Identification of γ-H2AX+ cells in lung parenchyma (white arrow) with IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (h) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. The scale bars in right lower corners represent 20 μm. Blue fluorescence indicates DAPI-stained nuclei. Statistical analysis in (d) and (h) using one-way ANOVA, followed by Bonferroni multiple comparison post hoc test. Symbols (∗, †, ‡) indicate significance (at 0.05 level).

Mentions: IF staining revealed that the number of γ-H2AX+ cells in medial layer of large PA and in lung parenchyma (i.e., extra-PA), an index of DNA damage, was significantly higher in group 2 than that in groups 1 and 3 and higher in group 3 than that in group 1 (Figure 3). Besides, IF staining of Ki-67+ cells (i.e., double staining of K-i67 and α-smooth muscle actin) (Figures 4(a)–4(d)) for quantifying SMC proliferation in the medial layer of large PA showed a pattern similar to that of γ-H2AX+ cells among the three groups. Furthermore, IF staining for identifying TRPC1 (i.e., double staining of TRPC1 and α-smooth muscle actin) (Figures 4(e)–4(h)) in the medial layer of large PA showed a prevalence of TRPC1+ cells, an index of hypoxia-inducible biomarker, similar to that of γ-H2AX+ cells among the three groups.


Reducing TRPC1 Expression through Liposome-Mediated siRNA Delivery Markedly Attenuates Hypoxia-Induced Pulmonary Arterial Hypertension in a Murine Model.

Sun CK, Zhen YY, Lu HI, Sung PH, Chang LT, Tsai TH, Sheu JJ, Chen YL, Chua S, Chang HW, Chen YL, Lee FY, Yip HK - Stem Cells Int (2014)

Immunofluorescent (IF) staining for γ-H2AX+ cells in medial layer of pulmonary arterioles (PA) and lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) (n = 10). ((a) to (c)) Identification of γ-H2AX+ cells in PA (white arrow) using IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (d) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. ((e) to (g)) Identification of γ-H2AX+ cells in lung parenchyma (white arrow) with IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (h) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. The scale bars in right lower corners represent 20 μm. Blue fluorescence indicates DAPI-stained nuclei. Statistical analysis in (d) and (h) using one-way ANOVA, followed by Bonferroni multiple comparison post hoc test. Symbols (∗, †, ‡) indicate significance (at 0.05 level).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Immunofluorescent (IF) staining for γ-H2AX+ cells in medial layer of pulmonary arterioles (PA) and lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) (n = 10). ((a) to (c)) Identification of γ-H2AX+ cells in PA (white arrow) using IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (d) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. ((e) to (g)) Identification of γ-H2AX+ cells in lung parenchyma (white arrow) with IF double staining (i.e., α-SMA-γ-H2AX) in the three groups (400x). (h) ∗ versus other groups with different symbols (∗, †, ‡), P < 0.0001. The scale bars in right lower corners represent 20 μm. Blue fluorescence indicates DAPI-stained nuclei. Statistical analysis in (d) and (h) using one-way ANOVA, followed by Bonferroni multiple comparison post hoc test. Symbols (∗, †, ‡) indicate significance (at 0.05 level).
Mentions: IF staining revealed that the number of γ-H2AX+ cells in medial layer of large PA and in lung parenchyma (i.e., extra-PA), an index of DNA damage, was significantly higher in group 2 than that in groups 1 and 3 and higher in group 3 than that in group 1 (Figure 3). Besides, IF staining of Ki-67+ cells (i.e., double staining of K-i67 and α-smooth muscle actin) (Figures 4(a)–4(d)) for quantifying SMC proliferation in the medial layer of large PA showed a pattern similar to that of γ-H2AX+ cells among the three groups. Furthermore, IF staining for identifying TRPC1 (i.e., double staining of TRPC1 and α-smooth muscle actin) (Figures 4(e)–4(h)) in the medial layer of large PA showed a prevalence of TRPC1+ cells, an index of hypoxia-inducible biomarker, similar to that of γ-H2AX+ cells among the three groups.

Bottom Line: By day 28, right ventricular systolic pressure (RVSP), number of muscularized arteries, right ventricle (RV), and lung weights were increased in group 2 than in group 1 and reduced in group 3 compared with group 2.The mRNA expressions of proapoptotic and hypertrophy biomarkers displayed a similar pattern, whereas sarcomere length showed an opposite pattern compared to that of RVSP in all groups.Lipofectamine siRNA delivery effectively reduced TRPC1 expression, thereby attenuating PAH-associated RV and pulmonary arteriolar remodeling.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency Medicine, E-Da Hospital, I-Shou University College of Medicine, Kaohsiung 82445, Taiwan.

ABSTRACT
We tested the hypothesis that Lipofectamine siRNA delivery to deplete transient receptor potential cation channel (TRPC) 1 protein expression can suppress hypoxia-induced pulmonary arterial hypertension (PAH) in mice. Adult male C57BL/6 mice were equally divided into group 1 (normal controls), group 2 (hypoxia), and group 3 (hypoxia + siRNA TRPC1). By day 28, right ventricular systolic pressure (RVSP), number of muscularized arteries, right ventricle (RV), and lung weights were increased in group 2 than in group 1 and reduced in group 3 compared with group 2. Pulmonary crowded score showed similar pattern, whereas number of alveolar sacs exhibited an opposite pattern compared to that of RVSP in all groups. Protein expressions of TRPCs, HIF-1α, Ku-70, apoptosis, and fibrosis and pulmonary mRNA expressions of inflammatory markers were similar pattern, whereas protein expressions of antifibrosis and VEGF were opposite to the pattern of RVSP. Cellular markers of pulmonary DNA damage, repair, and smooth muscle proliferation exhibited a pattern similar to that of RVSP. The mRNA expressions of proapoptotic and hypertrophy biomarkers displayed a similar pattern, whereas sarcomere length showed an opposite pattern compared to that of RVSP in all groups. Lipofectamine siRNA delivery effectively reduced TRPC1 expression, thereby attenuating PAH-associated RV and pulmonary arteriolar remodeling.

No MeSH data available.


Related in: MedlinePlus