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Keratinocyte Growth Factor Gene Electroporation into Skeletal Muscle as a Novel Gene Therapeutic Approach for Elastase-Induced Pulmonary Emphysema in Mice.

Tobinaga S, Matsumoto K, Nagayasu T, Furukawa K, Abo T, Yamasaki N, Tsuchiya T, Miyazaki T, Koji T - Acta Histochem Cytochem (2015)

Bottom Line: In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells.Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection.Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice.

View Article: PubMed Central - PubMed

Affiliation: Division of Surgical Oncology, Department of Translational Medical Sciences, Nagasaki University Graduate School of Biomedical Sciences , Nagasaki, Japan.

ABSTRACT
Pulmonary emphysema is a progressive disease with airspace destruction and an effective therapy is needed. Keratinocyte growth factor (KGF) promotes pulmonary epithelial proliferation and has the potential to induce lung regeneration. The aim of this study was to determine the possibility of using KGF gene therapy for treatment of a mouse emphysema model induced by porcine pancreatic elastase (PPE). Eight-week-old BALB/c male mice treated with intra-tracheal PPE administration were transfected with 80 μg of a recombinant human KGF (rhKGF)-expressing FLAG-CMV14 plasmid (pKGF-FLAG gene), or with the pFLAG gene expressing plasmid as a control, into the quadriceps muscle by electroporation. In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells. Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection. Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice. These results indicated that KGF gene therapy with electroporation stimulated lung epithelial proliferation and protected depression of pulmonary function in a mouse emphysema model, suggesting a possible method of treating pulmonary emphysema.

No MeSH data available.


Related in: MedlinePlus

Histological changes in mouse lung after instillation of PPE with or without KGF gene transfection. Lung tissues of mice treated PPE were fixed on day 2 (A), day 7 (B), and day 28 (C) after instillation of PPE and were stained with hematoxylin and eosin. Alveolar wall destruction and air space enlargement were seen in a time-dependent manner. Lungs of mice on day 3 after PPE instillation and transfection with the pKGF-FLAG gene (D) or with the pFLAG gene (E) were similarly analyzed. Infiltration of inflammatory cells and hemorrhage were also seen in these tissues. Data are representative of 3 mice per group. Magnification: A, B and C (×40), Bar=200 μm; D and E (×200), Bar=100 μm.
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Figure 3: Histological changes in mouse lung after instillation of PPE with or without KGF gene transfection. Lung tissues of mice treated PPE were fixed on day 2 (A), day 7 (B), and day 28 (C) after instillation of PPE and were stained with hematoxylin and eosin. Alveolar wall destruction and air space enlargement were seen in a time-dependent manner. Lungs of mice on day 3 after PPE instillation and transfection with the pKGF-FLAG gene (D) or with the pFLAG gene (E) were similarly analyzed. Infiltration of inflammatory cells and hemorrhage were also seen in these tissues. Data are representative of 3 mice per group. Magnification: A, B and C (×40), Bar=200 μm; D and E (×200), Bar=100 μm.

Mentions: Initially, we characterized histological changes in the lung resulting from KGF gene transfection by using a PPE-induced emphysema model. In this model, as described previously, after intra-tracheal instillation of PPE, lung injury with progressive alveolar wall destruction and enlargement of air space was caused in a time-dependent manner. Infiltration of inflammatory cells diminished gradually after PPE instillation, and, by day 28 after PPE instillation, emphysema was almost established and alveolar destruction was completed (Fig. 3A, B and C). As shown in Fig. 3D and E, KGF-FLAG gene transfection immediately after PPE administration did not induce any changes in lung morphological details compared to FLAG gene transfection when analyzed three days after transfection.


Keratinocyte Growth Factor Gene Electroporation into Skeletal Muscle as a Novel Gene Therapeutic Approach for Elastase-Induced Pulmonary Emphysema in Mice.

Tobinaga S, Matsumoto K, Nagayasu T, Furukawa K, Abo T, Yamasaki N, Tsuchiya T, Miyazaki T, Koji T - Acta Histochem Cytochem (2015)

Histological changes in mouse lung after instillation of PPE with or without KGF gene transfection. Lung tissues of mice treated PPE were fixed on day 2 (A), day 7 (B), and day 28 (C) after instillation of PPE and were stained with hematoxylin and eosin. Alveolar wall destruction and air space enlargement were seen in a time-dependent manner. Lungs of mice on day 3 after PPE instillation and transfection with the pKGF-FLAG gene (D) or with the pFLAG gene (E) were similarly analyzed. Infiltration of inflammatory cells and hemorrhage were also seen in these tissues. Data are representative of 3 mice per group. Magnification: A, B and C (×40), Bar=200 μm; D and E (×200), Bar=100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: Histological changes in mouse lung after instillation of PPE with or without KGF gene transfection. Lung tissues of mice treated PPE were fixed on day 2 (A), day 7 (B), and day 28 (C) after instillation of PPE and were stained with hematoxylin and eosin. Alveolar wall destruction and air space enlargement were seen in a time-dependent manner. Lungs of mice on day 3 after PPE instillation and transfection with the pKGF-FLAG gene (D) or with the pFLAG gene (E) were similarly analyzed. Infiltration of inflammatory cells and hemorrhage were also seen in these tissues. Data are representative of 3 mice per group. Magnification: A, B and C (×40), Bar=200 μm; D and E (×200), Bar=100 μm.
Mentions: Initially, we characterized histological changes in the lung resulting from KGF gene transfection by using a PPE-induced emphysema model. In this model, as described previously, after intra-tracheal instillation of PPE, lung injury with progressive alveolar wall destruction and enlargement of air space was caused in a time-dependent manner. Infiltration of inflammatory cells diminished gradually after PPE instillation, and, by day 28 after PPE instillation, emphysema was almost established and alveolar destruction was completed (Fig. 3A, B and C). As shown in Fig. 3D and E, KGF-FLAG gene transfection immediately after PPE administration did not induce any changes in lung morphological details compared to FLAG gene transfection when analyzed three days after transfection.

Bottom Line: In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells.Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection.Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice.

View Article: PubMed Central - PubMed

Affiliation: Division of Surgical Oncology, Department of Translational Medical Sciences, Nagasaki University Graduate School of Biomedical Sciences , Nagasaki, Japan.

ABSTRACT
Pulmonary emphysema is a progressive disease with airspace destruction and an effective therapy is needed. Keratinocyte growth factor (KGF) promotes pulmonary epithelial proliferation and has the potential to induce lung regeneration. The aim of this study was to determine the possibility of using KGF gene therapy for treatment of a mouse emphysema model induced by porcine pancreatic elastase (PPE). Eight-week-old BALB/c male mice treated with intra-tracheal PPE administration were transfected with 80 μg of a recombinant human KGF (rhKGF)-expressing FLAG-CMV14 plasmid (pKGF-FLAG gene), or with the pFLAG gene expressing plasmid as a control, into the quadriceps muscle by electroporation. In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells. Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection. Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice. These results indicated that KGF gene therapy with electroporation stimulated lung epithelial proliferation and protected depression of pulmonary function in a mouse emphysema model, suggesting a possible method of treating pulmonary emphysema.

No MeSH data available.


Related in: MedlinePlus