Preventive And Therapeutic Effects Of Danshensu On Hypoxic Pulmonary Hypertension And Its Mechanism

Part one Preventive and Therapeutic effects of Danshensu and paeonol on hypoxic pulmonary hypertension.Background: hypoxic pulmonary hypertension(HPH)is a common respiratory disease,mainly increased pulmonary vascular resistance,pulmonary vascular remodeling,pulmonary artery pressure increased progressively.So far,there is no specific medicine to cure pulmonary hypertension.Proliferation of pulmonary artery smooth muscle cells(pulmonary arterial smooth musle cell,PASMC)play an important role in the pathophysiology of HPH,and anti-proliferation of smooth muscle cells,some drugs in animal experiments has demonstrated a good therapeutic effect on HPH.Traditional Chinese medicine is composed of Salvia miltiorrhiza and moutan recipes for traditional Chinese medicine preparation standard,one of the main active ingredients of Salvia Danshensu could protect the vascular endothelial function,promote vascular diastolic function,The main effective components of paeonol,anti-inflammatory,antioxidation,anti hypertension and other aspects of the role,but the combined use of Danshensu and paeonol is effective in the prevention and treatment of HPH in Denmark,has not been studied yet.This topic will be through the establishment of HPH animal model to study the preliminary danshensu Pae Pae and danshensu used in conjunction with inhibition and possible mechanisms of HPH.Objective: To explore the preventive and therapeutic effects of danshensu(DSS)and paeonol(Pae)on hypoxic pulmonary hypertension in rats.Method: SD rats were randomly divided into the following groups: normoxia group,hypoxia group,prevention group and treatment group with different dose of danshensu and paeonol.The right ventricular pressure(the mean pulmonary artery pressure),right ventricular hypertrophy index and mean left common carotid artery pressure were measured and compared.Results: Prophylactic administration of two doses of 160 mg/kg and 320 mg/kg of danshensu can significantly reduce right ventricular pressure and right ventricular hypertrophy index in hypoxic rats,effectively preventing the development of HPH,however,there was no significant difference between the two effective doses.No significant effect was seen when the therapeutic administration of different doses of Danshensu.Different doses of paeonol had no significant effect on the prevention of HPH in rats.The combination administration had an effect on the prevention of HPH in rats.Summary: Prophylactic administration of 160 mg/kg and 320 mg/kg danshensu can significantly reduce right ventricular pressure and right ventricular hypertrophy index in hypoxic rats,thus effectively preventing the occurrence of hypoxic pulmonary hypertension in rats.Part two Effect and mechanism of danshensu on pulmonary vascular remodeling in HPHBackground: HPH is a continued contraction in the pulmonary artery,pulmonary artery pressure and structural remodeling characterized by a progressive increase in a common respiratory disease,The excessive proliferation of pulmonary artery smooth muscle cells(PASMCS)caused by hypoxia or transformation of transforming(factor-β,tgf-β),which eventually leads to pulmonary remodeling,is an important mechanism for the formation of HPH.Danshensu(Dan Shen Su),is extracted from Salvia miltiorrhiza in a water-soluble phenolic compound,a large number of studies have shown that expansion of coronary Danshensu,anticoagulant,anti-myocardial ischemia and hypoxia,anti-cell hyperproliferation,a variety of protection against myocardial remodeling and other cardiovascular,But danshensu whether inhibition of pulmonary arterial remodeling HPH,so far there is no relevant reports.In this study,we will establish an animal model of hypoxic pulmonary hypertension to investigate the inhibitory effect of tanshinone on HPH pulmonary vascular remodeling and its possible mechanism.Objective: To explore the preventive effect and possible mechanism of Danshensu on hypoxic pulmonary remodeling.Method: SD rats were randomly divided into the following groups:(1)normoxia group;(2)normoxia+Danshensu group;(3)hypoxic group;(4)hypoxic+Danshensu group.Intraperitoneal injection of Danshensu(160 mg/kg daily)or normal saline in different groups.Pulmonary arterial histology,pulmonary arterial hemodynamics,and right ventricular hypertrophy index were measured in each group,qRT-PCR technique was used to detect the change of mRNA expression of TGF-β and Smad3 in lung tissue of rats in each group.Results: Danshensu can decrease the induction of hypoxia on pulmonary vascular remodeling in rats.Danshensu significantly inhibited the mRNA expression level of TGF-β in hypoxia group and the protein expression of TGF-β and p-Smad3 in lung tissue.Summary: Danshensu inhibits pulmonary artery remodeling and pulmonary artery pressure increasing in HPH rats.Part three Mechanism and Inhibitory Effect of Danshensu on Proliferation of Pulmonary Artery Smooth Muscle CellsBackground: HPH is a common respiratory disease caused by hypoxia or TGF-β artery smooth muscle cells(of PASMCs)hyperproliferative,it plays a more important role in the pathophysiology of HPH.Danshensu could inhibit HPH proliferation of pulmonary artery smooth muscle cells,there is no related reports,This topic will be isolated and cultured in vitro pulmonary artery smooth muscle cells by pulmonary vascular adventitial fibroblasts(pulmonary artery fibroblasts PAFs)and alveolar macrophage cell line NR8383 Preliminary studies at the cellular level by HPH Salvia Inhibition effects on pulmonary artery smooth muscle cell proliferation and its possible mechanism.Objective: To explore the effect of Danshensu on the proliferation of pulmonary artery smooth muscle cells and its mechanism.Method: Rat pulmonary artery smooth muscle cells(PASMCs),pulmonary artery adventitial fibroblasts(PAFs)and NR8383 were cultured and given different treatments depending on the experimental design.A trypan blue exclusion test was used to explore the safe dose of danshensu at the cellular level.The effect of Danshensu on proliferation of PASMC was observed by MTT assay combined with cell counting.The expression of TGF-β in NR8383 and PAFs was detected by qRT-PCR.ELISA was utilized to detect the secretion of TGF-β from NR8383 and PAFs.Western blot was used to detect the effect of Danshensu on the expression of TGF-β and Smad3.Results: 30μg/ml danshensu could significantly inhibit hypoxia-induced proliferation of PASMC without affecting the activity of normal cells.30μg/ml danshensu significantly inhibited the hypoxia-induced synthesis and secretion of TGF-βinduction in NR8383 and PAFs.30μg/ml Danshensu markdely inhibited the proliferation of PASMCs cultured under hypoxia and TGF-β conditions.Danshensu could decrease the phosphorylation level of p-Smad3 induced by TGF-β,but had no significant effect on the protein expression of total Smad3.Summary: Danshensu can effectively inhibit hypoxia-induced proliferation of PASMCs.Reduce the secretion of TGF-β,in PAFs and NR8383 cells under hypoxic conditions.At the same time,Danshensu can significantly inhibit the expression of TGF-β and the phosphorylation of Smad3 in PASMCs under hypoxic conditions.Conclusion: Prophylactic administration of Danshensu can improve right ventricular remodeling and pulmonary arterial remodeling in hypoxic rats to prevent hypoxic pulmonary hypertension in rats.The mechanism may be that Danshensu can be achieved by regulating the TGF-β-Smad3 pathway and inhibiting hypoxia-induced proliferation of PASMCs.