Tetrandrine prevents monocrotaline-induced pulmonary arterial hypertension in rats through regulation of the protein expression of inducible nitric oxide.

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Tetrandrine prevents monocrotaline-induced pulmonary arterial hypertension in rats through regulation of the protein expression of inducible nitric oxide synthase and cyclic guanosine monophosphate-dependent protein kinase type 1 Xiaowu Wang, PhD, Yongchao Yang, MD, Dongpeng Yang, MD, Guang Tong, PhD, Shanshan Lv, MD, Xi Lin, PhD, Changfu Chen, MD, Wenpeng Dong, PhD Journal of Vascular Surgery Volume 64, Issue 5, Pages 1468-1477 (November 2016) DOI: 10.1016/j.jvs.2015.09.016 Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 1 a, Rats injected with monocrotaline (MCT) developed pulmonary hypertension, as shown by increases in the right ventricular (RV) systolic pressure at 42 days after injection. Tetrandrine (Tet) treatment (80 mg/kg/d from day 21 to day 42, intraperitoneal [IP]) prevented development of pulmonary hypertension, comparable to vardenafil (Val) treatment (2 mg/kg/d from day 21 to day 42, IP). b, Adaptive RV hypertrophy was demonstrated by right ventricle weight: right ventricle/left ventricle plus septum (RV/LV + S). Tet treatment prevented RV hypertrophy. Data are expressed as mean ± standard error of the mean (n = 10-12 rats). ∗P < .05 vs control; #P < .05 vs MCT. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 2 a-h, Compared with normal rats, monocrotaline (MCT)-injected rats demonstrated significant increase of medial wall thickness, stenosis of small pulmonary arteries, monocyte infiltration, inflammatory cell infiltration, and disorganized collagen fiber proliferation. Tetrandrine (Tet)-treated rats showed an obvious improvement in all aspects described before, similar to vardenafil (Val)-treated rats (magnification ×200). CON, Control. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 3 a-d, Density of middle membrane smooth muscle cells (SMCs), pulmonary artery media thickness (PAMT), percentage of wall thickness with outer diameter (WT%), and percentage of wall area with total area (WA%) were measured as indices of proliferation of pulmonary artery SMCs (PASMCs). Data are shown as mean ± standard error of the mean. ∗P < .05 vs control; #P < .05 vs monocrotaline (MCT). Tet, Tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 4 a and b, The average diameter (AD) of myocardial cells and myocardial nucleus density (MND) were the indices of right ventricular (RV) remodeling. Data are shown as mean ± standard error of the mean. ∗P < .05 vs control; #P < .05 vs monocrotaline (MCT). Tet, Tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 5 a-d, Electron microscopy micrographs show the difference between different groups in the morphology of endothelial cells, vacuoles, and organelles, especially outstretched endoplasmic reticulum (arrow) and increased Golgi apparatus (magnification ×30,000). Con, Control; MCT, monocrotaline; Tet, tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 6 a-d, The expression of inducible nitric oxide synthase (iNOS) protein was shown in the endothelial cells of pulmonary arteries and epithelium in the alveolar wall. The arrow indicates the pulmonary vascular. Positive expression is dark brown (magnification ×200). Con, Control; MCT, monocrotaline; Tet, tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 7 a-d, The immunostaining of protein kinase type 1 (PKG-1) was shown in the pulmonary artery walls, bronchial walls, and airway epithelium. The arrow indicates the pulmonary vascular (a, b, d) and airway epithelium (c). Positive expression is dark brown (magnification ×200). Con, Control; MCT, monocrotaline; Tet, tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 8 a and b, Protein expression of inducible nitric oxide synthase (iNOS) and cyclic guanosine monophosphate-dependent protein kinase type 1 (PKG-1), assessed by Western blotting, in homogenate of lung tissue from control, monocrotaline (MCT)-treated, tetrandrine (Tet)-treated, and vardenafil (Val)-treated rats. β-actin was used for normalization. Data are shown as mean ± standard error of the mean. ∗P < .05 vs control; #P < .05 vs MCT. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 9 a-d, The expression of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and catalase (CAT) in homogenate of lung tissue in different groups (21 days after Tet treatment). Data are represented as mean ± standard error of the mean. ∗P < .05 vs control; #P < .05 vs monocrotaline (MCT). Tet, Tetrandrine; Val, vardenafil. Journal of Vascular Surgery 2016 64, 1468-1477DOI: (10.1016/j.jvs.2015.09.016) Copyright © 2015 Society for Vascular Surgery Terms and Conditions