Protective Effects Of Aloperine On Pulmonary Artery Hypertension

Objectives Observe the protective effects of Aloperine on pulmonary artery hypertension in rats and explore its mechanism.Methods Right cardiac catheterization was used to detect hemodynamic parameters in pulmonary arterial hypertension in rats;H.E.staining was adopted to observe the changes of the heart and lung morphology of pulmonary arterial hypertension in rats;Masson staining was adopted to observe the growth of collagenous fibers in lung tissue of pulmonary arterial hypertension in rats;Transmission electron microscopy was adopted to observe the changes of pulmonary vascular endothelial cells in pulmonary arterial hypertension in rats;Immunohistochemical staining was used to observe the changes of intercellular adhesion molecule-1?MCP-1?and endothelin?ET-1?expression in lung tissues of pulmonary arterial hypertension in rats;Western blot was used to detect the changes in the expression levels of NF-?B,TNF-?and IL-1?in pulmonary arterial hypertension in rats;CCK-8 method was used to detect the cytotoxicity of Aloperine in HPASMCs;CCK-8 method and BrdU method were used to test the effects of Aloperine on the proliferation and DNA synthesis of HPASMCs;Flow cytometry was used to determine the cell cycle and apoptosis of HPASMCs;Western blot was used to detect the changes in the expression levels of IKK??I?B??p-I?B??p-IKK??NF-?B?TNF-??p27^kip1?Cyclin E1 in HPASMCs;Results The hemodynamic results showed that compared with the control group,the aloperine?50,100mg/kg?group could significantly reduce the mPAP,RVSP and RVHI of the pulmonary arterial hypertension compared with the model group;In the model group,the results of H.E.staining of the lung tissue showed the arterial wall was significantly thickened and the vascular cavity was narrow.Compared with the model group,treat with aloperine can reverse the remodeling of pulmonary artery in rats.Right ventricular myocardial tissue H.E.staining,according to the results in control group,myocardial arrangement was disordered,shape was irregular,fiber hyperplasia and myocardial cell swelling,myocardial cell diameter?AD?and nucleus density?MND?increased significantly in rats in model group;The myocardial tissue arrangement in the treatment group was orderly,and the cell swelling and irregular morphology were improved,which significantly reduced the AD and MND of myocardial cells in aloperine group compared with the model group;Masson staining results showed that,compared with the control group,the collagen fibers were significantly hyperplasia,and the muscle fibers were irregular in the lung tissues in model group.Compared with the model group,the treatment can significantly improve the collagen fiber hyperplasia in lung tissues of pulmonary hypertension in rats,and the muscle fibers are arranged in a continuous and orderly manner;The results of transmission electron microscopy showed that compared with the control group,the endothelial cells in the model group showed obvious proliferation and expansion,morphological changes,swelling of the endoplasmic reticulum and golgi body;Compared with the model group,the damage of endothelial cells in the Aloperine?100mg/kg?group was improved and the swelling of the organelle was alleviated;In the model group,immunohistochemical results showed that the expression of MCP-1 and ET-1 in the lung tissues of was significantly increased.The treatment of Aloperine?100mg/kg?could reduce the expression of MCP-1 and ET-1 in the lung tissues of rats and improve the inflammatory response in the lungs;In the model group,western blot results showed that compared with the control group,the expression levels of NF-?B,TNF-?and IL-1?in the lung tissues of were significantly increased.The treatment of aloperine?100mg/kg?was able to significantly reduce the expression level of NF-?B,TNF-?and IL-1?;The results of CCK-8 and BrdU showed that aloperine?0.25,0.5mM?had inhibitory effect on the proliferation and DNA synthesis in HPASMCs.Cell cycle detection by flow cytometry found that aloperine has inhibitory effect on the proliferation of HPASMCs induced by PDGF-BB and promotes apoptosis;Western blot results showed that aloperine could inhibit the expression of IKK??I?B??p-I?B??p-IKK??NF-?B?TNF-??p27^kip1?Cyclin E1 in HPASMCs induced by PDGF-BB,and the expression of p27^kip1 protein was increased,and no effect was found on the expression of IKK and I?B?.Conclusion 1.The aloperine has protective effect on the pulmonary arterial hypertension induced by monocrotaline in rats.2.The aloperine has effects in inhibiting proliferation of HPASMCs induced by PDGF-BB and promoting its apoptosis.3.It may be possible to protect the pulmonary arterial hypertension by NF-?B signaling pathway.