Effects of Polydatin on Proliferation and Apoptosis of Pulmonary Artery Smooth Muscle Cells in Neonatal Rats with Hypoxic Pulmonary Hypertension by Regulating YAP1/TAZ Signaling Pathway / 华中科技大学学报(医学版)

Objective To investigate the effects of polydatin(PD)on the proliferation and apoptosis of pulmonary artery smooth muscle cells(PASMCs)in hypoxic pulmonary hypertension(HPH)neonatal rats,and its mechanism of action.Methods Neonatal rats were randomly separated into six groups:control group,model group,low dose PD group,medium dose PD group,high dose PD group,and high dose PD+Hippo pathway inhibitor(high dose PD+XMU-MP-1)group,with 10 rats in each group.After 2 weeks of hypoxia treatment,the right ventricular systolic blood pressure(RVSP)and right ventricular hypertro-phy index(RVHI)of rats in each group were measured.Hematoxylin-eosin(HE)staining was applied to observe pathological changes in lung tissue,and the percentage of pulmonary artery wall thickness to total thickness(WT)and the percentage of wall area to total area(WA)were calculated.Neonatal rat PASMCs were separated from each group,which were divided into NC group,hypoxia group,low dose PD group,medium dose PD group,high dose PD group,and high dose PD+XMU-MP-1 group.Cell counting kit 8(CCK-8)and 5-ethynyl-2'-deoxyuridine(EdU)were applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Western blot was applied to detect the expression of Yes-associated protein 1(YAP1),tran-scriptional coactivator with PDZ-binding motif(TAZ),mammalian sterile 20-like kinase 1(MST1),B-cell lymphoma 2(Bcl-2),and Bcl-2 associated protein(Bax)in lung tissue and PASMCs.Results Compared with the control group,the pulmonary artery wall in the model group was significantly thickened,lumen was narrowed,and protein expressions of RVSP,RVHI,WT％,WA％,YAP1,MST1 and TAZ were significantly increased(all P＜0.05).Compared with the model group,pulmonary artery thickening and lumen enlargement were observed in the low,medium and high dose PD groups,and the protein expressions of RVSP,RVHI,WT％,WA％,YAP1,MST1 and TAZ were significantly decreased,which showed a dose-dependent relationship(all P＜0.05).The effect could be reversed by XMU-MP-1.Compared with the NC group,the cell A450nm value,EdU positive rate,the protein expression of YAP1,MST1,TAZ and Bcl-2 in the hydropoxia group were significantly increased.The apoptosis rate and the expression of Bax protein were obviously reduced(all P＜0.05).Compared with the hypoxia group,the cell A450nm value,EdU positive rate,the protein expression of YAP1,MST1,TAZ and Bcl-2 in the low,medium and high dose PD groups were obviously reduced.The apoptosis rate and the expression of Bax were significantly increased,which showed a dose-depend-ent relationship(all P＜0.05).The effect could be reversed by XMU-MP-1.Conclusion PD may inhibit the proliferation of PASMCs in HPH neonatal rats and promote apoptosis by inhibiting YAP1/TAZ signaling pathway.

Basic Research Progress of Zang Medicine in Treatment of Hypoxic Pulmonary Hypertension / 医药导报

Hypoxic pulmonary hypertension(HPH)is a kind of chronic high-altitude sickness disease,which is also the initiation link of high-altitude sickness such as pneumocardial disease and high-altitude pulmonary edema.Its main features are persistent vasoconstriction and irreversible remodeling of blood vessels.HPH has greatly impacted the health of people in the plat-eau section.As a typical representative of ethnic medicine,Zang medicine embodies the wisdom of the Zang people in their long-term struggle against diseases.Adapting to local conditions for a long time,Zang medicine has unique advantages in the treatment of HPH.By reviewing the pathogenesis of HPH and the research of single Zang medicine and the complex prescription of Zang medicine in the treatment of HPH,this article aims to provide a theoretical basis for the physiological and pathological research of HPH and to provide a reference for the clinical application of Zang medicine.

Investigation of Efficacy and Mechanism of Zukamu Granules on Hypoxic Pulmonary Hypertension / 医药导报

Objective To explore and verify the protective and therapeutic effects and possible mechanisms of Zukamu granules on hypoxia alone and hypoxia+Su5416-induced hypoxic pulmonary hypertension(HPH)in mice.Methods Multiple databases and related literature were used to collect the active ingredients data in Zukamu granules and the HPH-related targets were predicted and obtained.The network construction and enrichment analysis were performed.The HPH mouse models were es-tablished by two-week hypoxia and four-week hypoxia+Su5416 induction,and the relevant indicators and the main pharmacodyna-mic indexes such as right ventricular pressure were tested.Masson staining was used to observe the pathological changes in lung tissues,and Western blotting was used to detect the expression levels of bax,bcl-2,PI3K,p-PI3K,eNOS,and HIF-1α in lung tis-sues.Results A total of 167 active ingredients of Zukamu granules were screened,with 179 intersecting targets with HPH,in-cluding targets like PIK3CA and HIF-1.The validation experimental results showed that Zukamu granules could significantly re-duce right ventricular systolic pressure and right ventricular hypertrophy in HPH mice,and down-regulate the expression of bcl-2 and HIF-1α and up-regulate the expression of bax,PI3K,p-PI3K and eNOS in mice lung tissues.Conclusion Zukamu gran-ules may act against HPH by modulating bax/bcl and PI3K-eNOS/HIF-1α signaling pathways.

Mechanism of levosimendan in treating hypoxic pulmonary hypertension based on network pharmacology and molecular docking technology / 中国药理学通报

Aim To explore the efficacy of levosimendan on hypoxia pulmonary hypertension through animal experiments, and to further explore the potential mechanism of action using network pharmacological methods and molecular docking technique. Methods The rat model of hypoxia pulmonary hypertension was constructed to detect right heart systolic pressure and right heart remodeling index. HE , Masson, and VG staining were core targets were screened out. GO and KEGG pathway enrichment analysis were performed using the DAVID database. Molecular docking of the core targets was performed with the AutoDock software. Results The results of animal experiments showed that levosimendan had obvious therapeutic effect on hypoxia pulmonary hypertension. The network pharmacology results showed that SRC, HSP90AA1, MAPK1, PIK3R1, AKT1, HRAS, MAPK14, LCK, EGFR and ESR1 used to analyze the changes of rat lung histopathology. Search the Swiss Target Prediction, DrugBank Online, BatMan, Targetnet, SEA, and PharmMapper databases were used to screen for drug targets. Disease targets were retrieved from the GeneCards, OMIM databases. The "drug-target-disease" network was constructed after identification of the two intersection targets. The protein interaction network was constructed and the were the key targets to play a therapeutic role. Molecular docking showed good docking of levosimendan with all the top five core targets with degree values. Conclusions Levosimendan may exert a therapeutic effect on hypoxia-induced pulmonary hypertension through multiple targets.

The preventive effect and mechanism of Dayuanyin in hypoxic pulmonary hypertension through NF-κB signaling pathway / 药学学报

Dayuanyin (DYY) has been shown to reduce lung inflammation in both coronavirus disease 2019 (COVID-19) and lung injury. This experiment was designed to investigate the efficacy and mechanism of action of DYY against hypoxic pulmonary hypertension (HPH) and to evaluate the effect of DYY on the protection of lung function. Animal welfare and experimental procedures are approved and in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Science. Male C57/BL6J mice were randomly divided into 4 groups: control group, model group, DYY group (800 mg·kg-1), and positive control sildenafil group (100 mg·kg-1). The animals were given control solvents or drugs by gavage three days in advance. On day 4, the animals in the model group, DYY group and sildenafil group were kept in a hypoxic chamber containing 10% ± 0.5% oxygen, and the animals in the control group were kept in a normal environment, and the control solvent or drugs continued to be given continuously for 14 days. The right ventricular systolic pressure, right ventricular hypertrophy index, organ indices and other metrics were measured in the experimental endpoints. Meantime, the expression levels of the inflammatory factors in mice lung tissues were measured. The potential therapeutic targets of DYY on pulmonary hypertension were predicted using network pharmacology, the expression of nuclear factor kappa B (NF-κB) signaling pathway-related proteins were measured by Western blot assay. It was found that DYY significantly reduced the right ventricular systolic pressure, attenuated lung injury and decreased the expression of inflammatory factors in mice. It can also inhibit hypoxia-induced activation of NF-κB signaling pathway. DYY has a protective effect on lung function, as demonstrated by DYY has good efficacy in HPH, and preventive administration can slow down the disease progression, and its mechanism may be related to inhibit the activation of NF-κB and signal transducer and activator of transcription 3 (STAT3) by DYY.

Effects of vascular endothelial growth factor inhibitor SU5416 on pulmonary vascular remodeling in neonatal rats with hypoxic pulmonary hypertension / 中国新生儿科杂志

Objective:To study the effects of vascular endothelial growth factor (VEGF) inhibitor SU5416 on pulmonary vascular remodeling in neonatal rats with hypoxic pulmonary hypertension (HPH).Methods:Thirty-two neonatal rats (age 7~10 d) were randomly assigned into normoxia group, hypoxia group, SU5416+hypoxia group and SU5416+normoxia group according to corresponding treatments. The endpoint was 14 d after treatments. Right ventricular systolic pressure (RVSP) was measured and cardiac and pulmonary tissue were sampled on the day. Right ventricular hypertrophy index (RVHI) was calculated. After HE staining, the morphological changes of pulmonary vessels were observed under light microscope.Media thickness (MT), external diameter(ED), cross-sectional area of tunica media (MA) and total cross-sectional area(TA) of pulmonary arterioles were measured. MT%(MT/ED) and MA%(MA/TA) were calculated as indicators of pulmonary vascular remodeling. The levels of VEGF and α-smooth muscle actin (α-SMA) in pulmonary vessels were examined using immunohistochemistry (IHC) method.Results:RVSP, RVHI, MT% and MA% in hypoxia group and SU5416+hypoxia group were significantly higher than normoxia group and SU5416+normoxia group ( P<0.05). These indicators in SU5416+hypoxia group were also higher than hypoxia group ( P<0.05),and no significant differences existed between SU5416+normoxia group and normoxia group ( P>0.05). The levels of α-SMA and VEGF in pulmonary vessels in SU5416+hypoxia group and hypoxia group were significantly higher than normoxia group and SU5416+normoxia group ( P<0.05), and α-SMA and VEGF in SU5416+hypoxia group higher than hypoxia group ( P<0.05). Conclusions:VEGF inhibitor SU5416 may exacerbate pulmonary hypertension and increase pulmonary vascular remodeling in neonatal HPH rats.

Hypoxia-preconditioned hUCMSC-Exos relieve hypoxic pulmonary hy-pertension by inhibiting pulmonary vascular EndMT / 中国病理生理杂志

AIM:To investigate the effect of hypoxia-preconditioned human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSC-Exos)on pulmonary vascular endothelial-mesenchymal transition(EndMT)in hypoxic pulmonary hypertension(HPH).METHODS:(1)Primary hUCMSCs were isolated and cultured by tissue adhesion method,and hUCMSC-Exos were extracted by ultrafiltration and identified.(2)Twenty-four SPF male SD rats were ran-domly divided into normoxia(N)group,hypoxia(H)group,hypoxia+normoxic hUCMSC-Exos group and hypoxia+hypoxia-preconditioned hUCMSC-Exos group,with 6 rats in each group.The rats in H group and intervention groups were placed in a cabin that simulated the hypoxic environment at an altitude of 5 000 m,and normoxic hUCMSC-Exos,hypoxia-precon-ditioned hUCMSC-Exos or equivalent volume of PBS were injected through the tail vein on the 3rd,5th,7th,10th and 14th days in hypoxia environment.After 21 d of modeling,the right ventricular systolic pressure(RVSP)and right ven-tricular hypertrophy index(RVHI)of the rats were detected,and the pathological changes of lung tissues were observed by HE staining.(3)After starvation for 12 h,human pulmonary arteriole endothelial cells(HPAECs)were randomly di-vided into normoxic control(N-Con)group,hypoxic model(H-Con)group,hypoxia+normoxic hUCMSC-Exos group and hypoxia+hypoxia-preconditioned hUCMSC-Exos group.The migration ability and tube formation ability of HPAECs were detected by Transwell assay and tube formation experiment.The expression of CD31 and α-smooth muscle actin(α-SMA)in HPAECs was detected by immunofluorescence double-staining.The protein levels of CD31,VE-cadherin,α-SMA and vimentin in pulmonary vessels and HPAECs were assessed by Western blot.RESULTS:(1)The HPH rat model was suc-cessfully established after 21 d of hypoxia,and EndMT occurred in pulmonary vessels.Compared with N group,the levels of RVSP,RVHI,percentage of vascular wall area(WA%)and percentage of vascular wall thickness(WT%)in H group were significantly increased(P＜0.01),pulmonary vascular wall thickening and the protein levels of CD31 and VE-cad-herin were significantly decreased(P＜0.01),while the protein levels of α-SMA and vimentin were significantly increased in pulmonary vessels(P＜0.05 or P＜0.01).Compared with H group,the RVSP,RVHI,WA%and WT%(P＜0.01)were significantly decreased(P＜0.05 or P＜0.01),and pulmonary vascular remodeling was attenuated after normoxic or hypoxia-preconditioned hUCMSC-Exos intervention.After hypoxia-preconditioned hUCMSC-Exos intervention,HPH pul-monary vascular remodeling and EndMT formation were significantly inhibited.(2)After 48 h of hypoxic treatment,the migration,tubule formation and EndMT of HPAECs were induced.Compared with H-Con group,cell migration and tube formation were significantly decreased after hypoxia-preconditioned hUCMSC-Exos intervention(P＜0.01).The protein levels of CD31 and VE-cadherin were increased,while the protein levels of α-SMA and vimentin were decreased(P＜0.05 or P＜0.01).CONCLUSION:Hypoxia-preconditioned hUCMSC-Exos attenuate the formation of HPH pulmonary vascu-lar remodeling by inhibiting pulmonary vascular EndMT.

Progress on the mechanism of PDGF-BB signaling pathway in the pulmonary vascular remodeling of neonatal hypoxic pulmonary hypertension / 国际儿科学杂志

Neonatal hypoxic pulmonary hypertension(HPH)is a common acute critical disease in NICU, and is one of the diseases leading to neonatal death.At present, the specific pathogenesis is still unclear.Current studies have shown that pulmonary vascular remodeling is an important pathological feature of pulmonary hypertension, and the excessive proliferation and migration of pulmonary artery smooth muscle cell is the main cause of pulmonary vascular remodeling.Platelet-derived growth factor(PDGF-BB)is a powerful mitogenic factor which involved in cell proliferation and migration.Currently, plenty of studies have found that PDGF-BB plays an important role in multiple diseases, including tumor, atherosclerosis, pulmonary hypertension and pulmonary fibrosis.In view of the mechanism of PDGF-BB, this article reviews the possible mechanism of PDGF-BB in pulmonary vascular remodeling with neonatal HPH, aiming to provide a new direction for the therapies of reversing pulmonary vascular remodeling with neonatal HPH.

Research progress on mechanisms and therapeutic drugs of hypoxic pulmonary hypertension / 中国药理学通报

Hypoxic pulmonary hypertension ( HPH) is a complex mechanism of HPH is complex, and it has a high mortality rate cardiopulmonary disease eaused by hypoxia.The pathological of disability.Clinically the diug of treatment for HPH is unspe-cialized, mainly relying on traditional vasomotor dnrgs, inclu¬ding prostaglandin 12 receptor agonists, endothelin receptor an¬tagonists and phosphodiesterase-5 inhibitors, but their efficacy cannot be achieved.To meet the clinical need, it is of great sig¬nificance to develop targeted anti-HPH dnigs.To provide ideas for the discovery of HPH treatment drugs, the pathophysiological mechanism of HPH and the current status of dmg development are reviewed in the paper.

Research progress on hypoxia-induced imbalance of calcium homeostasis in pulmonary artery smooth muscle cells and its treatment / 中国药理学通报

Chronic hypoxic lung diseases are major causes of disability and mortality worldwide, which are typically aggravated by hypoxic pulmonary hypertension.The pathogenesis of hypoxic pulmonary hypertension is complex, and its mechanism has not been fully elucidated.The previous studies have shown abnormally elevated levels of free Ca + in the cytoplasm of pulmonary artery smooth muscle cells to be the predominant drivers of pulmonary hypertension , causing continuous contraction and remodeling of the pulmonary vessels.This article briefly summarizes the mechanism of hypoxia-induced imbalance in calcium homeostasis in pulmonary artery smooth muscle cells, together with its related drug research, based on the existing literature.Hypoxia induces an imbalance in calcium homeostasis in pulmonary artery smooth muscle cells by regulating hypoxia-inducible factor-1, K+ , store-operated calcium channel, receptor-operated calcium channel, the Ca +-sensing myosin contractile mechanism by binding to calmodulin, leading to pulmonary vasoconstriction.Ca + can also activate PKC/ MAPKs and PI3K/Akt/mTOR pathways, leading to pulmonary vascular remodeling.

Calcium signal transduction in mitochondria in hypoxic pulmonary hypertension / 中国药理学通报

Hypoxic pulmonary hypertension (HPH) is characterized by hypoxic pulmonary vasoconstriction (H P V) and hypoxic pulmonary vascular remodeling(HPVR). Previous studies have shown that intracellular Ca

Maxing xiongting mixture regulates factors relevant to lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats / 中国组织工程研究

BACKGROUND: Hypoxic pulmonary hypertension is a key link in the progression from chronic obstructive pulmonary disease to cor pulmonale. Its severity is closely related to disease development and prognosis. Current treatments cannot prevent or reverse disease progression. Maxing Xiongting Mixture has significant effect on hypoxic pulmonary hypertension with the syndrome of intermingled phlegm and blood stasis. OBJECTIVE: To study how the Maxing Xiongting Mixture regulates relevant factors of lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis. METHODS: Seventy Sprague-Dawley rats, 5 weeks old, were randomly divided into normal group (n=10) and model group (n=60), where acute cor pulmonale model was prepared by injecting 50 mg/kg monocrotaline solution (1%) intraperitoneally, followed by forced smoking and swimming 6 days a week lasting for 4 weeks. Except for 10 rats in the normal group, there were 46 model rats in the model group. According to the normal distribution of body mass, 40 rats were selected and randomly divided into 4 groups: model group, high-dose Maxing Xiongting Mixture group (MH), low-dose Maxing Xiongting Mixture group (ML) and fasudil group, with 10 rats in each group. Rats in MH and ML groups were respectively given Maxing Xiongting Mixture at 20 g/(kg·d) and 5 g/(kg·d), respectively and those in the fasudil group were given fasudil at a dose of 10 mg/(kg·d). Other groups were given equal amount of saline. Administration was given intraperitoneally and intragastrically, once a day for 14 days in total. RT-PCR was used to test the expression of factors related to lung reshaping and vascular remodeling, including RhoA, stromelysin 1 and tumor necrosis factor-α mRNAs. An approval for the study was obtained from the Ethics Committee of Chengdu University of Traditional Chinese Medicine (approval No. 2017-03). RESULTS AND CONCLUSION: Compared with the model group, the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs were significantly lowered in the MH group (all P 0.05). To conclude, Maxing Xiongting Mixture, which is similar to fasudil, intervenes lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis by inhibiting the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs.

MicroRNA-143–5p modulates pulmonary artery smooth muscle cells functions in hypoxic pulmonary hypertension through targeting HIF-1α

This paper explores the potential mechanism of microRNA-143–5p regulation effects on pulmonary arterysmooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a,which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143–5pmimics or inhibitor control/miR-143–5p inhibitor. We used Western blotting and RT-qPCR to detect the proteinand mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellularmigration measurement and Dual luciferase reporter gene assay to prove the target of miR-143–5p. Cells underhypoxic condition presented the decreased protein and mRNA expressions of a-smooth muscle actin (SM-aactin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22a (SM22a),Calponin1 and Hypoxia-inducible factor-1a(HIF-1a), the increased cell viability and miR-143–5p level; Overexpression of miR-143–5p obviously reduced vascular smooth muscle-specific contraction marker proteinlevels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143–5p caused the opposite changes, while co-transfected with Si HIF-1a blocked thebeneficial effects of miR-143–5p inhibition on PASMCs under hypoxia. MicroRNA-143–5p can promote thephenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxiccondition through direct targeting of HIF-1a.

Clinical Observation of Bosentan in the Treatment of Hypoxic Pulmonary Hypertension in Neonates / 中国药房

OBJECTIVE：To observe the ef fectiveness and safety of bosentan in the treatment of hypoxic pulmonary hypertension（HPH）in neonates. METHODS ：From Jan. 2014 to Mar. 2019，a total of 82 HPH neonates hospitalized in the department of neonatology of our hospital were selected as research subjects. According to whether or not receiving bosentan therapy，50 cases were included into bosentan group and 32 cases into non-bosentan group. Meanwhile ，another 25 non-HPH neonates with serum sample retention time and general information such as gestational age at birth and day age matching the HPH group were selected as the control group. All neonates with HPH were given continuous intravenous infusion of Dopamine hydrochloride injection 5 mg/（kg·min）until PASP was normal. On this basis, neonates in the bosentan group were additionally given Bosentan tablets 1 mg/kg（fed after dissolving with appropriate amount of water for injection ）for q 12 h，72 h. The relationship between serum ET- 1 levels of neonates with HPH and PASP was analyzed ，as well as PASP before and after treatment and therapeutic efficacy between bosentan and non-bosentan groups ，the changes of arterial blood gas indexes and ADR in 3 groups were compared. RESULTS ：Before treatment ，the serum ET- 1 levels of bosentan group was （164.3±115.3）pg/mL，which was significantly higher than （41.9±3.7）pg/mL of control group and positively correlated with PASP level （r＝0.864，P＜0.001）. Total response rate of bosentan group was 90.00％，which was significantly higher than 71.88％ of non-bosentan group （P＜0.05）. After 72 h of treatment ，PASP of 2 groups was decreased significantly ，compared with before treatment （P＜0.001），and the bosentan group was significantly lower than the non-bosentan group （P＜0.05）. The PaO 2，SaO2，PaCO2 and OI in 3 groups was significantly improved compared with that before treatment （P＜0.001），and the PaO 2，SaO2 and OI in the bosentan group was significantly higher than that in the non-bosentan group （P＜0.05）. During the treatment period of bosentan and within one week after drug withdrawal，there was no significant change in serum LDH ，AST，ALT and Scr levels in neonates. There was no statistically significant difference in the incidence of feeding intolerance ，anemia，reduced WBC and reduced PLT in 3 groups（P＞0.05）. CONCLUSIONS： Bosentan can improve the oxygenation status of neonates with HPH, reduce PA SP，and short-termmedication is safe. com

Role of Ca2+ in hypoxic pulmonary vasoconstriction / 中华危重病急救医学

In acute hypoxia, pulmonary vascular will contract and divert blood to better ventilated area to optimize ventilation/perfusion matching, which is known as hypoxic pulmonary vasoconstriction (HPV). In chronic hypoxia, irreversible pulmonary vascular remodeling can be induced, characterized by pulmonary artery middle smooth muscle cells and the outer fiber cell hyperplasia in luminal stenosis and pulmonary artery hypertension (PAH) eventually. Furthermore, PAH can cause increased ventricular afterload, and right heart failure in severe cases. Pulmonary artery smooth muscle cell (PASMC) elevated Ca2+ concentration is one of the most important factors of its contractions, proliferation and migration. Recent studies on Ca2+ promoting in HPV were summarized in order to provide evidence for clinical prevention of hypoxia and therapeutic PAH.

17β-estradiol Attenuates Hypoxia-induced Pulmonary Vascular Remodeling via Downregulating miRNA-21 Signaling Pathway / 中国循环杂志

Objectives: To explore the effect of 17β-estradiol (E2) on hypoxic pulmonary hypertension (HPH) and explore if the effects were mediated through suppressing pulmonary artery smooth muscle cells (PASMCs) proliferation by targeting miRNA-21 (miR-21). Methods: Animal experiment: A total of 32 healthy female SD rats with castrated surgery were randomly divided into 4 groups: normoxia group, normoxia+E2 group, hypoxia group, hypoxia+E2 group (n=8 each). The rats in normoxia+E2 group and hypoxia+E2 group received subcutaneous injection of E2 20 μg/kg/d, and the rest groups received subcutaneous injection of equal volume saline. The hypoxic groups were placed in the hypoxic chamber (24 hours per day for 8 weeks) to establish HPH model and normoxic groups were kept in the room air. The pulmonary artery remodeling, mean pulmonary artery pressure (mPAP), right ventricle hypertrophy index (RVHI) were observed. Real-time PCR and Western blot were used to detect the levels of proliferation cell nuclear antibody (PCNA) and miR-21 expression in pulmonary artery. In vitro: human pulmonary artery smooth muscle cells (hPASMCs) were randomly divided into 3 groups: normoxia group, hypoxia group, hypoxia+E2 group. The levels of cell proliferation in each group were tested by MTT after 24 hours. Real-time PCR and Western blot were used to detect the levels of PCNA and miR-21 in cells. Results: Animal experiment: compared with normoxia group, the hypoxia group showed obviously thickened pulmonary artery wall, increased mPAP and RVHI, and significantly increased expression of miR-21 and PCNA (P<0.01);above changed were significantly attenuated in hypoxia+E2 group (P<0.01). In vitro: compared with normoxia group, the hypoxia group showed obvious proliferation and significantly increased expression of miR-21 and PCNA (P<0.01);compared with hypoxia group, the proliferation of hPASMCs and expression of miR-21 and PCNA were obviously reduced in hypoxia+E2 group (P<0.01). Conclusions: E2 could effectively reduce mPAP, attenuate the degree of right heart hypertrophy and pulmonary vascular remodeling, the protective effect may be mediated through downregulating miR-21 and PCNA expression, and subsequently inhibiting the proliferation of hPASMCs.

Protective effect of salidroside on rats with hypoxic pulmonary hyperten-sion by suppressing oxidative stress / 中国病理生理杂志

AIM:To study whether salidroside plays a protective role in hypoxia-induced pulmonary hyperten-sion by suppressing oxidative stress.METHODS: Sprague-Dawley rats were randomly divided into 4 groups: normoxia (N)group,hypoxia for 4 weeks(H4)group,low-dose salidroside(hypoxia for 4 weeks and treatment with salidroside at 16 mg/kg,H4S16)group and high-dose salidroside(hypoxia for 4 weeks and treatment with salidroside at 32 mg/kg, H4S32)group.The mean pulmonary arterial pressure(mPAP), the weight ratio of right ventricle/(left ventricle+sep-tum)[RV/(LV+S)]and vessel wall area/vessel total area(WA/TA)were evaluated.The levels of malondialdehyde (MDA)in the serum and lung tissues were detected by colorimetric method.The levels of 8-iso-prostaglandin F2α(8-iso-PGF2α)in the serum and lung tissues were measured by ELISA.The activity of superoxide dismutase(SOD)in the serum was analyzed by hydroxylamine method.The expression of NAPDH oxidase 4(NOX4)and SOD1 in the lung tissues was determined by Western blot.RESULTS:Compared with N group,the levels of mPAP,RV/(LV+S)and WA/TA in H4 group were significantly increased,which were apparently attenuated by salidroside injection in a dose-dependent manner. Meanwhile,salidroside administration apparently decreased the levels of MDA and 8-iso-PGF2αin the serum and lung tis-sues,as well as the expression of NOX 4 in the lung tissues.Besides,compared with N group, the activity of SOD in the serum and the expression of SOD1 in the lung tissues in H4group were significantly decreased,while administration of sali-droside increased the activity of SOD in the serum and the expression of SOD 1 in the lung tissues in a dose-dependent man-ner.CONCLUSION:Salidroside protects the pulmonary vessels from remodeling and attenuates hypoxia -induced pulmo-nary hypertension by inhibiting oxidative stress.

Protective effect of heat shock protein 70 on lungs in newborn rats with hypoxic pulmonary hypertension / 中华实用儿科临床杂志

Objective To investigate the protective effect of adenovirus mediated heat shock protein 70 (HSP70) on lungs in neonatal rats with hypoxic pulmonary hypertension (HPH).Methods One hundred and twenty-eight 7-10 d healthy Wistar neonatal rats were randomly divided into HPH model group and control group.HPH group was divided into saline group,empty virus group,and HSP70 group according to the transfection solution.HPH model was established in the hypoxia cabin of 80 mL/L nitrogen oxygen mixed gas after transfection.The mean pulmonary artery pressure(mPAP) was measured after 3,7,10 and 14 days of hypoxia in each group.The mRNA and protein expression of HSP70,hypoxia inducible factor-1 alpha(HIF-1 α),endothelin-1 (ET-1) and inducible nitric oxide synthase(iNOS) in the lung tissues of neonatal rats were detected by using reverse transcription-PCR and Western blot respectively.Results (1) The mPAP level was significantly higher in saline group (M,Q:12.00,2.50;15.00,2.00;18.00,1.75;20.00,2.25) than that in control group (M,Q:9.50,4.75;10.50,1.00;13.00,1.00;15.50,3.25),and the differences were significant (z =-3.28,-3.40,-3.34,-3.06,all P ＜ 0.01);and the differences were also significant between empty virus group (M,Q:13.50,2.00;15.50,1.75;18.00,1.00;22.00,4.25) and control group (z =-2.83,-3.42,-3.40,-2.97,all P ＜ 0.01) in 3,7,10,and 14 days;but there was no significant difference between HSP70 group (M,Q:8.50,4.00;10.50,1.00;13.00,1.00)and the control group in 3,7,and 10 days (z =-0.43-0.00,-3.06,all P ＞ 0.05).(2) The expressions of HSP70 mRNA among the groups were statistically significant(F =6.321,9.669,6.333,all P ＜ 0.01),and the expressions of HSP70 protein also had significant difference(F =16.463,3.637,17.749,all P ＜ 0.01).(3)The level of HIF-1α mRNA in saline group was significantly higher than that of the control group,and the differences were statistically significant (q =4.312,9.106,6.151,all P ＜ 0.01);and the level of HIF-1α mRNA in empty virus group was also significantly higher than that in the control group,and the differences were statistically significant (q =3.982,9.235,5.352,all P ＜ 0.01) in 3,7,and 10 days;hypoxia in HSP70 group was lower than that of the empty virus group in 3,7 days,and the differences were statistically significant (q =6.083,11.031,all P ＜ 0.05).The level of ET-1 mRNA in saline group was significantly higher than that in the control group(q =5.112,10.086,6.264,all P ＜ 0.01),in empty virus group was significantly higher than that in the control group,and the differences were statistically significant (q =4.182,12.238,5.864,all P＜0.01) in 3,7,and 10 days,but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =6.912,10.235,7.021,all P ＜ 0.05).The level of iNOS mRNA in saline group was significantly higher than that of the control group,and the differences were statistically significant (q =4.998,8.056,5.369,all P ＜0.01),in empty virus group was significantly higher than that in the control group,and the differences were statistically significant (q =4.778,10.138,5.154,all P ＜0.01) in 3,7,and 10 days,but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =7.819,9.838,6.156,all P ＜ 0.05).The level of HIF-1 α protein in saline group was significantly higher than that of the control group in 3,7,and 10 days,and the differences were statistically significant (q =3.146,3.012,4.106,all P ＜ 0.05),in empty virus group was significantly higher than that of the control group in 10 days,and the difference was statistically significant (q =3.468,P ＜ 0.05);but in HSP70 group it was lower than that in the empty virus group in 3,7,and 10 days,and the differences were statistically significant (q =3.876,4.108,4.021,all P＜ 0.05).The level of ET-1 protein of HSP70 group was lower than that of the saline group,the differences were statistically significant(q =3.367,2.983,3.246,all P ＜ 0.05),in HSP70 group was lower than that of the empty virus,and the differences were statistically significant (q =3.268,2.678,3.567,all P ＜0.05).The level of iNOS protein in saline group was significantly higher than that in the control group in 3,7,and 10 days,and the differences were statistically significant (q =3.360,3.567,3.567,all P ＜ 0.05),but in HSP70 group it was lower than that in the empty virus group,and the differences were statistically significant (q =3.126,3.908,3.087,all P ＜ 0.05).Conclusion Adenovirus mediated HSP70 can improve the HSP70 expression in HPH,down-regulate the expression of HIF-1 α,ET-1,iNOS,and reduce pulmonary arterial pressure.

Effect of PASMC apoptosis on reversal of hypoxic pulmonary arterial remodeling during reoxygenation and its related molecular mechanism / 中国病理生理杂志

AIM: To explore the effect of pulmonary arterial smooth muscle cell (PASMC) apoptosis on the reversal of hypoxic pulmonary arterial remodeling during reoxygenation and its possible mechanism.METHODS: Male SD rats (n=24) were randomly divided into normoxia for 4 weeks group, hypoxia for 4 weeks group, reoxygenation for 1 week after hypoxia for 4 weeks group and reoxygenation for 6 weeks after hypoxia for 4 weeks group.Right ventricular systolic pressure (RVSP), right ventricular hypertrophy index, pulmonary arterial medial thickness (MT) and medial area (MA) as well as autophagy and apoptosis in the pulmonary arterial medial layer were examined during hypoxia-reoxygenation.The rat primary PASMCs were divided into normoxia for 48 h group, hypoxia for 48 h group, reoxygenation for 24 h after hypoxia for 48 h group and normoxia for 72 h group to explore the changes of PASMC autophagy and apoptosis following hypoxia-reoxygenation.Finally, primary PASMCs were divided into normoxia for 72 h group, reoxygenation for 24 h after hypoxia for 48 h group and reoxygenation for 24 h after hypoxia for 48 h + chloroquine (inhibitor of autophagy) group to investigate the effect of PASMC autophagy during hypoxia on the apoptosis during reoxygenation.RESULTS: After hypoxia for 4 weeks, the RVSP, during right ventricular hypertrophy index, MT and MA increased significantly compared with normoxia group (P<0.05), and gradually decreased during reoxygenation.The expression of LC3 in the pulmonary arterial medial layer increased evidently after hypoxia and gradually reversed during reoxygenation.Moreover, the P62 and cleaved caspase-3 expression decreased after hypoxia compared with normoxia group, and increased markedly following reoxyge-nation.The expression of cleaved caspase-3/PARP in rat primary PASMCs decreased significantly under hypoxia (P<0.05), and increased evidently during reoxygenation.The expression of P62 and LC3-II decreased markedly under hypoxia (P<0.05).After inhibition of PASMC autophagy under hypoxia, the expression of cleaved caspase-3/PARP decreased remarkably during reoxygenation (P<0.05).CONCLUSION: The PASMC apoptosis participates in the reversal of hypoxic pulmonary arterial remodeling, and the PASMC autophagy under hypoxia might facilitate its apoptosis during reoxygenation.

Advances in differential roles of HIF-1αand HIF-2αin the pathogenesis of hypoxic pulmonary hypertension / 中国药理学通报

Hypoxia-induced factors(HIFs)are the main regula-tors for the response of hypoxic environment.They are involved in hypoxia-related lung tissue cell damage and abnormal cell pro-liferation,among which,HIF-1αand HIF-2αplay the most im-prominant roles.This paper reviews the current researches of HIF-1αand HIF-2α,focusing on their structural and functional similarities and diversities,as well as their roles in the patho-genesis of hypoxic pulmonary hypertension.