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.

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.

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.

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.

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.

The expression and significance of circulating microRNA of rats with hypobaric hypoxia-induced pulmonary hypertension / 西安交通大学学报(医学版)

Objective To investigate the expression of circulating microRNA (miRNA) of rats with hypobaric hypoxia‐induced pulmonary hypertension (HPH) .Methods Commercial rat miRNA microarray was employed to detect and analyze the circulating miRNA profile in the serum samples of Sprague‐Dawley rats with hypobaric hypoxia‐induced HPH and controls .Furthermore ,differentially expressed candidate circulating miRNAs between HPH and control groups were validated by Real‐time quantitative PCR based on the case‐control study ,and receiver operating characteristic curve (ROC ) analysis was used to test the performance of four differentially expressed circulating miRNAs in discriminating HPH and control groups .Results Compared with those in the control group ,13 upregulated miRNAs and 10 downregulated miRNAs were identified in hypobaric hypoxia‐induced HPH rats by using miRNA microarray . And differentially expressed miR‐451 , miR‐505 , let‐7d and miR‐214 were validated by using RT‐PCR .ROC analysis showed that the area under the curve of miR‐451 ,miR‐505 and let‐7d was 0 .979 ,0 .938 and 0 .993 in discriminating HPH and control groups ,respectively .Conclusion The aberrant expression of circulating miR‐451 ,miR‐505 and let‐7d in serum may be correlated with the pathogenesis of HPH .

Changes and significance of endothelin-1 in lung tissue of hypoxic pulmonary hypertension development in neonatal rats / 中华实用儿科临床杂志

Objective To know about the effect and significance of endothelin-1 (ET-1) in pathogenesis of hypoxic pulmonary hypertension(HPH) in neonatal rats.Methods The 120 newborn Wistar rats were randomly divided into hypoxic group and control group,and the rats of hypoxic group were made into HPH model.Mean pulmonary arterial pressure(mPAP) was measured on day 3,5,7,10,14 and 21 after hypoxia.The morphological change of pulmonary vessels were observed,and pulmonary vascular remodeling indexes were also obtained,including the ratio of middle wall thickness to external diameter of small pulmonary arteries (MT％) and the ratio of media cross-section area to total cross-sectional area of small pulmonary arteries(MA％).Immunohistochemistry and Western blot tests were done to determine the expression of ET-1 in lung tissue.Results 1.After hypoxia for 3,5,7,10,14,21 days,the levels of mPAP in hypoxia group[(8.492 ± 1.548) mm Hg,(10.022 ± 1.182) mm Hg,(11.470 ± 2.868) mm Hg,(16.842 ±2.154) mm Hg,(12.824 ± 2.859) mm Hg,(21.036 ± 2.590) mm Hg; 1 mm Hg =0.133 kPa] were significantly higher than those in control group[(5.141 ± 1.022) mm Hg,(8.137 ± 1.057) mm Hg,(8.730 ±0.868) mm Hg,(12.125 ±2.541) mm Hg,(8.920 ±0.744) mm Hg,(11.156 ±1.644) mm Hg] (all P ＜0.05).2.Seven days after hypoxia exposure,small pulmonary arterials remodeling was observed.MT％ [(53 ± 11) ％] and MA％ [(60 ± 9) ％]in hypoxia group were also significantly higher than those in control group [(49 ± 11) ％,(54 ± 8) ％] (all P ＜ 0.05).3.The results of immunohistochemistry:the expression intensities of ET-1 were significantly higher in hypoxia group (0.614,0.613,0.651) after hypoxia for 3,5,7 days than those in control group (0.433,0.386,0.369) (all P ＜ 0.05).4.The results of Western blot:the levels of ET-1 were significantly higher in hypoxia group(4.885 ± 1.391,4.434 ± 1.726,6.309 ± 0.330,2.353 ± 0.961) after hypoxia for 3,5,7 and 10 days than those in control group (1.698 ± 0.794,1.454 ± 0.776,2.045 ± 0.668,0.766 ± 0.515) (all P ＜ 0.05).5.The results of correlation analysis:the total expression of ET-1 protein was positively correlated with the total level of mPAP on 3-7 days after hypoxia (r =0.459,P ＜ 0.05).However,the expression of ET-1 had no significant correlation with MT％ and MA％ (rMT =-0.041,rMA =0.322,all P ＞ 0.05).Conclusions The level of ET-1 in lung tissue of HPH newborn rats increased in the early stage after hypoxia exposure and no longer increased during the late stage,which indicated that hypoxia tolerance prevailed.ET-1 acted as an early reactive marker in the process of HPH development in neonatal rats and may be mainly associated with elevated mPAP,which further prompted that through valid means to inhibit the expression of ET-1 in the early stage of hypoxia,in which the prevention and treatment of this disease may be more effective.

Expression of NO and H_2S in hypoxic pulmonary hypertension and intervention of sodium nitrite in rats / 中国病理生理杂志

AIM:To observe the change of nitric oxide(NO)and hydrogen sulfide(H_2S)in blood and lung homogenate of hypoxic pulmonary hypertension(HPH)rat model,and to discuss the meaning of inhalation sodium nitrite and these factors in the treatment of HPH. METHODS:Fifty healthy male Wistar rats were assigned randomly into 5 groups(10 rats each):normoxia control group(NC),normoxia sodium nitrite group(NNI),hypoxic control group(HC),hypoxic normal saline group(HNS)and hypoxic sodium nitrite group(HNI). The mean pulmonary arterial pressure(mPAP),weight of right ventricle,weight of left ventricle plus septum,and the ratio of the weight of right ventricle to that of left ventricle plus septum(right ventricle hypertrophy index,RVHI)were also determined. The serum level of NO and plasma level of H_2S were measured,and at the same time the levels of NO in the lung homogenate were detected. The structures in pulmonary arteries were examined using optical microscope. RESULTS:After model established,compared to that in the normoxia groups,the body weight decreased significantly in hypoxia groups(P<0.05),although no difference of body weight in five groups before producing model was observed. Compared to that in normoxia groups,the levels of mPAP and RVHI increased significantly in hypoxia groups,and compared to that in hypoxia control groups and hypoxia normal saline group,mPAP and RVHI levels decreased significantly in hypoxia sodium nitrite group(P<0.05). Compared to that in normoxia groups,the serum level of NO decreased significantly in hypoxia groups(P<0.05). NO level in lung homogenate decreased significantly in hypoxia control group and hypoxia normal saline group as compared to that in normoxia groups(P<0.05),and no obvious difference between hypoxic sodium nitrite group and normoxia groups was found. The plasma level of H_2S was decreased significantly in hypoxia groups(P<0.05)as compared to that in normoxia groups. H_2S level increased significantly in hypoxia sodium nitrite group as compared to that in hypoxia control groups and hypoxia normal saline group(P<0.05). Observation under optical microscope,the lumen structure of lung in normoxia control group was normal. No significant change in normoxia sodium nitrite group was found. The proliferation of smooth muscle cells(SMCs),the collagen fiber deposition in the vessel wall and every caliber thickening was observed in hypoxic control group. The same changes were also observed in hypoxic normal saline group. The thickened caliber was relieved significantly in hypoxic nitrite group. CONCLUSION:Pulmonary hypertension and right ventricle reconstitution can be relieved by inhalation of sodium nitrite,and can be regulated by the level of NO and H_2S in rats. Above all,inhalation of sodium nitrite may degrade HPH directly or by affecting the externalization and synthesizing of gas signaling molecule indirectly.

Effects of iptakalim on the mRNA and protein expression of eNOS in pulmonary tissue of chronic hypoxic rats / 中国药理学通报

Aim To study mRNA and protein expression of eNOS in pulmonary tissue of chronic hypoxic pulmonary hypertension(HPH)rats and chronic hypoxic rats treated with novel KATP opener iptakalim.Methods sixty Sprague-Dawley(SD)male rats were randomly divided into control group, hypoxic group, low dose iptakalim group(0.75 mg·kg~(-1)·d~(-1)), and high dose iptakalim group(1.5 mg·kg~(-1)·d~(-1)).Except the first group, the other three groups were put into hypoxic and normobaric chamber (10%±0.5% O_2,8 h/day and 6 day/week) to establish chronic hypoxic model. After four weeks, the mean pulmonary arterial pressure(mPAP), RV/(LV+S)and the plasma concentration of NO were measured. RT-PCR was performed to analyze the mRNA expression of eNOS in pulmonary tissue. Western blot was performed to analyze the protein expression of eNOS, iNOS in pulmonary tissue. Results ① The level of mPAP and RV/ ( LV + S) were significantly higher in the hypoxic group than those in control group ( P < 0. 05 ) , Low dose iptakalim groupandhighdoseiptakalimgroupdecreased the level of mPAP and RV/( LV + S) significantly (P <0. 05). ② The level of NO was significantly lower in the hypoxic group than those in control group (P<0. 05). Low dose iptakalim group and high dose iptakalim group increased the level of NO significantly (P < 0. 05 ). ③ The mRNA and protein expression of eNOS in the hypoxic group were significantly lower than those in the control group (P < 0. 05 ). Low dose iptakalim group and high dose iptakalim group increased the expression of eNOS significantly ( P < 0. 05). High dose iptakalim group was more significant. Conclusion Pulmonary vascular endothelial dysfunction is induced by chronic hypoxia,and the level of NO, the mRNA and protein expression of eNOS are decreased. Iptakalim can improve the vascular endothelial dysfunction, increase the expression of eNOS and the level of NO and reverse hypoxic pulmonary hypertension.

Expression of hypoxia-inducible factor-1? in the lungs of rats with hypoxic pulmonary hypertension / 中国呼吸与危重监护杂志

Objective To study expression of hypoxia inducible factor 1? (HIF 1?) in the lungs of hypoxic rats and to explore the role of HIF 1? in the pathogenesis of hypoxic pulmonary hypertension Methods Twenty Wistar rats were randomly divided into control group and hypoxia group The models of hypoxic pulmonary hypertension were established by exposure to hypoxia for 4 weeks according to our laboratory protocol Digoxin labelled cRNA probe for HIF 1? was prepared by in vitro transcription Northern blot was performed by using the HIF 1? cRNA probe and In situ hybridization was also conducted with rat lung tissue sections Results Northern blot hybridization showed minimally positive in the lung tissue of control group ,but strongly positive in hypoxia group In situ hybridization analysis with hypoxic rat lung tissue revealed that HIF 1? mRNA was expressed in bronchial epithelial cells (strongly positive reaction) and peribronchial proliferative lymphomatic tissue (weakly positive reaction),casually in alveolar wall cells (weakly positive reaction),but not in pulmonary arterial endothelium and smooth muscle cells Conclusions Our data suggested that chronic hypoxia could induce pulmonary hypertension characterized by pulmonary vascular remodeling HIF 1? mRNA expression was elevated in the lungs of rat under hypoxic condition HIF 1 may be involved in the pathophysiological changes in response to hypoxia and play an important role in the development of hypoxic pulmonary hypertension