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Recombinant adeno-associated virus-mediated human kallikrein gene therapy protects against hypertensive target organ injuries through inhibiting cell apoptosis.

Yan JT, Wang T, Wang DW - Acta Pharmacol. Sin. (2009)

Bottom Line: Overexpression of human tissue kallikrein (HK), mediated by recombinant adeno-associated virus (rAAV), decreased blood pressure in spontaneous hypertensive rats (SHRs) and reduced injury to the heart, aorta and kidney.Treatment with rAAV-HK decreased cell apoptosis in the target organs of SHRs and also inhibited lipopolysaccharide (LPS)-induced HEK 293 apoptosis.The rAAV-HK delivery system also increased the levels of apoptosis-inhibiting proteins bcl-2 and bcl-x(L), and decreased the level of Bax and the activity of caspase 3, two promoters of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine and the Institute of Hypertension, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.

ABSTRACT

Aim: Overexpression of human tissue kallikrein (HK), mediated by recombinant adeno-associated virus (rAAV), decreased blood pressure in spontaneous hypertensive rats (SHRs) and reduced injury to the heart, aorta and kidney. In this study, we used both an in vivo animal model and in vitro cell culture system to investigate whether rAAV-mediated HK gene therapy protects against organ damage by inhibiting cell apoptosis.

Methods: rAAV encoding HK (rAAV-HK) or LacZ (rAAV-lacZ) were delivered as a control to spontaneously hypertensive rats (SHRs) and cultured human embryonic kidney (HEK) 293 cells.

Results: Treatment with rAAV-HK decreased cell apoptosis in the target organs of SHRs and also inhibited lipopolysaccharide (LPS)-induced HEK 293 apoptosis. The rAAV-HK delivery system also increased the levels of apoptosis-inhibiting proteins bcl-2 and bcl-x(L), and decreased the level of Bax and the activity of caspase 3, two promoters of apoptosis. In addition to its role in the inhibition of apoptosis, rAAV-HK also activated the cell survival and proliferation signaling pathways ERK1/2 and PI3K/AKT.

Conclusion: rAAV-mediated HK gene delivery has multiple therapeutic possibilities for treating hypertension, not only by decreasing blood pressure, but also by directly inhibiting end-organ damage.

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Related in: MedlinePlus

The kallikrein/kinin system activates cell proliferation signaling pathways in HEK 293 cells. Western blot analysis was performed on cells in non-FBS medium exposed to LPS (A), and the relative expression levels of p-ERK1/2, P-AKT, and PI3K as determined by densitometry, were normalized to total ERK1/2, AKT, and β-actin, as shown in the histograms (B, C, D). The expression levels of PI3K, phospho-AKT and phospho-ERK1/2 decreased upon LPS exposure. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on the expression of PI3K, p-AKT and p-ERK1/2. Kinin+Hoe140 had no effects on the LPS-induced decrease in PI3K, phospho-AKT and phospho-ERK1/2 expression. bP<0.05, cP<0.01 vs positive control.
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fig7: The kallikrein/kinin system activates cell proliferation signaling pathways in HEK 293 cells. Western blot analysis was performed on cells in non-FBS medium exposed to LPS (A), and the relative expression levels of p-ERK1/2, P-AKT, and PI3K as determined by densitometry, were normalized to total ERK1/2, AKT, and β-actin, as shown in the histograms (B, C, D). The expression levels of PI3K, phospho-AKT and phospho-ERK1/2 decreased upon LPS exposure. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on the expression of PI3K, p-AKT and p-ERK1/2. Kinin+Hoe140 had no effects on the LPS-induced decrease in PI3K, phospho-AKT and phospho-ERK1/2 expression. bP<0.05, cP<0.01 vs positive control.

Mentions: We also confirmed that the kallikrein/kinin system activates the PI3K/AKT and ERK1/2 signaling pathways in HEK 293 cells using Western blotting. Expression of PI3K, phospho-AKT and phospho-ERK1/2 decreased when HEK 293 cells were exposed to LPS in non-FBS medium. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on PI3K, phospho-AKT and phospho-ERK1/2 expression (P<0.05). Kinin+Hoe140 did not counteract the decreased expression of PI3K, phospho-AKT and phospho-ERK1/2 induced by LPS (Figure 7). These data indicate that the kallikrein/kinin system activates the PI3K/AKT and ERK1/2 signaling pathways by activating the B2 receptor, which may contribute to the observed anti-apoptotic effects of the kallikrein/kinin system in HEK 293 cells.


Recombinant adeno-associated virus-mediated human kallikrein gene therapy protects against hypertensive target organ injuries through inhibiting cell apoptosis.

Yan JT, Wang T, Wang DW - Acta Pharmacol. Sin. (2009)

The kallikrein/kinin system activates cell proliferation signaling pathways in HEK 293 cells. Western blot analysis was performed on cells in non-FBS medium exposed to LPS (A), and the relative expression levels of p-ERK1/2, P-AKT, and PI3K as determined by densitometry, were normalized to total ERK1/2, AKT, and β-actin, as shown in the histograms (B, C, D). The expression levels of PI3K, phospho-AKT and phospho-ERK1/2 decreased upon LPS exposure. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on the expression of PI3K, p-AKT and p-ERK1/2. Kinin+Hoe140 had no effects on the LPS-induced decrease in PI3K, phospho-AKT and phospho-ERK1/2 expression. bP<0.05, cP<0.01 vs positive control.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4007182&req=5

fig7: The kallikrein/kinin system activates cell proliferation signaling pathways in HEK 293 cells. Western blot analysis was performed on cells in non-FBS medium exposed to LPS (A), and the relative expression levels of p-ERK1/2, P-AKT, and PI3K as determined by densitometry, were normalized to total ERK1/2, AKT, and β-actin, as shown in the histograms (B, C, D). The expression levels of PI3K, phospho-AKT and phospho-ERK1/2 decreased upon LPS exposure. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on the expression of PI3K, p-AKT and p-ERK1/2. Kinin+Hoe140 had no effects on the LPS-induced decrease in PI3K, phospho-AKT and phospho-ERK1/2 expression. bP<0.05, cP<0.01 vs positive control.
Mentions: We also confirmed that the kallikrein/kinin system activates the PI3K/AKT and ERK1/2 signaling pathways in HEK 293 cells using Western blotting. Expression of PI3K, phospho-AKT and phospho-ERK1/2 decreased when HEK 293 cells were exposed to LPS in non-FBS medium. Treatment with rAAV-HK and kinin, but not rAAV-lacZ, inhibited the effect of LPS on PI3K, phospho-AKT and phospho-ERK1/2 expression (P<0.05). Kinin+Hoe140 did not counteract the decreased expression of PI3K, phospho-AKT and phospho-ERK1/2 induced by LPS (Figure 7). These data indicate that the kallikrein/kinin system activates the PI3K/AKT and ERK1/2 signaling pathways by activating the B2 receptor, which may contribute to the observed anti-apoptotic effects of the kallikrein/kinin system in HEK 293 cells.

Bottom Line: Overexpression of human tissue kallikrein (HK), mediated by recombinant adeno-associated virus (rAAV), decreased blood pressure in spontaneous hypertensive rats (SHRs) and reduced injury to the heart, aorta and kidney.Treatment with rAAV-HK decreased cell apoptosis in the target organs of SHRs and also inhibited lipopolysaccharide (LPS)-induced HEK 293 apoptosis.The rAAV-HK delivery system also increased the levels of apoptosis-inhibiting proteins bcl-2 and bcl-x(L), and decreased the level of Bax and the activity of caspase 3, two promoters of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine and the Institute of Hypertension, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.

ABSTRACT

Aim: Overexpression of human tissue kallikrein (HK), mediated by recombinant adeno-associated virus (rAAV), decreased blood pressure in spontaneous hypertensive rats (SHRs) and reduced injury to the heart, aorta and kidney. In this study, we used both an in vivo animal model and in vitro cell culture system to investigate whether rAAV-mediated HK gene therapy protects against organ damage by inhibiting cell apoptosis.

Methods: rAAV encoding HK (rAAV-HK) or LacZ (rAAV-lacZ) were delivered as a control to spontaneously hypertensive rats (SHRs) and cultured human embryonic kidney (HEK) 293 cells.

Results: Treatment with rAAV-HK decreased cell apoptosis in the target organs of SHRs and also inhibited lipopolysaccharide (LPS)-induced HEK 293 apoptosis. The rAAV-HK delivery system also increased the levels of apoptosis-inhibiting proteins bcl-2 and bcl-x(L), and decreased the level of Bax and the activity of caspase 3, two promoters of apoptosis. In addition to its role in the inhibition of apoptosis, rAAV-HK also activated the cell survival and proliferation signaling pathways ERK1/2 and PI3K/AKT.

Conclusion: rAAV-mediated HK gene delivery has multiple therapeutic possibilities for treating hypertension, not only by decreasing blood pressure, but also by directly inhibiting end-organ damage.

Show MeSH
Related in: MedlinePlus