Transfection of mesenchymal stem cells (MSCs) with the mouse-secreted Klotho (mSKL) gene. A, Map of lentiviral vector expressing the mSKL and enhanced green fluorescent protein (eGFP) genes. B, Map of the control lentiviral vector expressing eGFP only. C, Phase-contrast image of MSCs in culture. D, eGFP expression in Lenti-SKL-GFP-transduced MSCs. E, Western blot analysis of SKL protein expression in cell lysates and conditioned medium from transfected and nontransfected MSCs. CMV indicates cytomegalovirus; cPPT, central polypurine tract; IRES, internal ribosome entry site; LTR, long terminal repeat; psi, Ψ domain; RRE, Rev responsive element; RSV, rous sarcoma virus; SIN, self-inactivating LTR as a result of the deletion of the U3 promoter; SRCMVi, a CMV promoter plus enhancers from RSV and SV40 promoters and a human β-globin intron; SV40, simian virus 40; and WPRE, woodchuck hepatitis virus post-transcriptional regulatory element.
Mesenchymal stem cells (MSCs) overexpressing secreted Klotho (SKL) abolished monocrotaline (MCT)-induced pulmonary vascular dysfunction. A, Responses of small pulmonary arteries (PAs) to ACh. B, Responses of small PAs to sodium nitroprusside. #P<0.05 vs the MCT group. n=6. Data=means±SEM. GFP indicates green fluorescent protein.
Mesenchymal stem cells (MSCs) overexpressing secreted Klotho (SKL) attenuated monocrotaline (MCT)-induced pulmonary arterial remodeling. A, Representative hematoxylin and eosin staining images of small pulmonary artery (PA) cross sections (60–80 µmol/L) in rat lungs. B, Quantitative analysis of medial thickness. C, Lumen area in rat small PA cross sections. D, α-Smooth muscle actin (α-SMA) staining using immunohistochemical in small PA cross sections. E, Quantification of α-SMA staining. Scale bar, 20 µm. *P<0.05 vs saline; #P<0.05 vs MCT. n=6. Data=means±SEM. GFP indicates green fluorescent protein.
Mesenchymal stem cells (MSCs) engrafted into the lung parenchyma and expressed secreted Klotho (SKL). A, Green fluorescent protein (GFP)-positive cells in the lungs of monocrotaline (MCT)+MSC-SKL-GFP and MCT+MSC-GFP groups. B, Western blot analysis of SKL protein levels in the lung lysates. *P<0.05 vs saline. n=6. Data=means±SEM.
Mesenchymal stem cells (MSCs) overexpressing secreted Klotho (SKL) attenuated monocrotaline (MCT)-induced inflammation in pulmonary arteries (PAs) and abolished downregulation of SIRT1 expression in the lungs. A, Immunohistochemical analysis of macrophages around small PAs (dark brown staining, CD68 marker). B, Quantification of CD68 staining. C, Western blot analysis of SIRT1 expression in lung lysates. *P<0.05, ***P<0.001 vs saline; #P<0.05 vs MCT. n=6. Data=means±SEM. GFP indicates green fluorescent protein; and SIRT1, Sirtuin 1.
Serum Klotho was decreased in patients with arterial stiffness and hypertension. A, Serum levels of Klotho. B, Brachial-ankle pulse wave velocity in hypertensive patients (HP) and normal controls (NC). C, Systolic blood pressure. D, Diastolic blood pressure in hypertensive patients and normal controls. n=13 to 14 participants/group. Data=means±SEM. *P<0.05, ***P<0.001 vs NC.
