肺动脉高压中内皮间质转化信号通路的研究进展

肺血管重塑是肺动脉高压(pulmonary arterial hypertension,PAH)主要的病理机制之一。内皮间质转化(endothelial to mesenchymal transition,EndMT)是肺血管重塑的重要病理基础。目前关于PAH机制的研究主要集中于血管细胞增殖,而涉及EndMT信号通路的研究并未深入。因此,探究肺动脉EndMT的信号机制并进行相应干预研究对治疗PAH起重要作用。     

野百合碱诱导大鼠肺动脉高压模型的建立

背景：目前尚缺乏简单易行、实用、操作性强的肺动脉高压动物模型。目的：建立一种实用的注射野百合碱诱导的肺动脉高压动物模型。方法：采用一次性皮下注射野百合碱60mg/kg的方法制备SD大鼠肺动脉高压模型。结果与结论：野百合碱注射后第1,2,3,4周,大鼠平均肺动脉压明显升高,右心室肥厚明显。光镜下可见肺小血管肌化程度增强,相对中膜厚度增加,肺血管密度减少,以上症状均随野百合碱注射时间的延长逐渐加重。证实此方法建立的大鼠肺动脉高压模型造模成功。     

野百合碱诱导大鼠肺动脉高压模型的建立

背景:目前尚缺乏简单易行、实用、操作性强的肺动脉高压动物模型.目的:建立一种实用的注射野百合碱诱导的肺动脉高压动物模型.方法:采用一次性皮下注射野百合碱60 mg/kg的方法制备SD大鼠肺动脉高压模型.结果与结论:野百合碱注射后第1,2,3,4周,大鼠平均肺动脉压明显升高,右心室肥厚明显.光镜下可见肺小血管肌化程度增强,相对中膜厚度增加,肺血管密度减少,以上症状均随野百合碱注射时间的延长逐渐加重.证实此方法建立的大鼠肺动脉高压模型造模成功.     

缬沙坦对野百合碱诱导肺动脉高压模型大鼠肺动脉重构的影响

目的:观察缬沙坦(valsartan)对野百合碱(monocrotaline)所致肺动脉高压大鼠肺血管重构的影响.方法:将健康雄性Wistar 大鼠随机分为3 组:M 组(肺动脉高压模型组)一次性项背部注射野百合碱(60 mg / kg)后自由摄食、饮水;V 组(缬沙坦干预组),同M 组注射野百合碱并同等条件饲养,4 周后开始用缬沙坦20 mg / (kg·d)灌胃,持续4 周达实验终点;C 组(正常对照组)一次性项背部注射等量生理盐水后与实验组同等条件饲养.然后经微导管介入测定大鼠肺动脉平均压(mPAP);计算右室肥大指数[RV/ (LV + S)];分别采用HE 染色、弹力纤维染色及VG 染色观察肺动脉结构的改变,计算肺动脉管壁厚度和管腔面积,评价缬沙坦对肺动脉重构的影响.结果:缬沙坦可有效降低野百合碱所致肺动脉高压大鼠模型肺动脉管壁的厚度,增大管腔面积(P ＜ 0.01).结论:缬沙坦可有效抑制肺动脉高压大鼠肺动脉重构,作用机制可能与其抑制AngⅡ介导的增殖效应有关.     

硝苯地平对野百合碱所致大鼠肺动脉高压的影响

目的探讨硝苯地平 (Nif)对野百合碱 (MCT)诱发慢性炎性肺动脉高压的防治作用。方法 3 3只雄性Wistar大鼠随机分为正常对照组、肺动脉高压模型组、Nif治疗组 ,每组 11只。予MCT (5 0mg/kg)制做大鼠肺动脉高压模型后 ,分别给Nif组及模型组大鼠连续灌胃Nif(2 0mg/kg·d)和等量生理盐水 2 1d。采用改良右心导管术测定肺血流动力学参数 ;处死大鼠后 ,称量肺湿重 (wW )、右心室自由壁 (RV)和左心室加室间隔 (LV S)重 ,计算右心肥厚指数 (RV/LV S)。结果Nif能明显降低肺动脉高压模型大鼠的平均肺动脉压及RV/LV S(P <0 0 1) ,但对wW无明显影响 (P >0 0 5 )。结论长期使用Nif能有效防治MCT所致肺动脉高压 ,改善心功能。     

野百合碱处理大鼠肺动脉高压模型的塑造

肺动脉高压尤其是特发性肺动脉高压（Idiopathic Pulrnonary Artery Hypertension, IPAH）是一种预后极差的疾病，其发病机制不明，治疗棘手。野百合碱（Monocrotatine，MCY）属于豆科植物野百合属，它所引起的肺动脉高压（Pulmonry Artery Hypertersion，PAH）大鼠是一种较为理想的肺动脉高压动物模型，在MCI-PAH动物模型大鼠血清中已发现内皮素-1（ET-1）、心钠素（ANP）及血栓素2（rXB2）明显升高，     

硫化氢逆转脂多糖诱导大鼠肺动脉低反应性以及与一氧化碳的关系

目的 观察整体水平应用硫化氢(H2S)后脂多糖(LPS)诱导的离体肺动脉对H2S舒张反应的变化及其与一氧化碳(CO)的关系.方法 将48只大鼠按照随机数字表法分为对照组[给予生理盐水(NS)]、LPS组、H2S供体硫氢化钠(NaHS)+LPS组和NaHS+NS组4组,每组12只.采用经大鼠气管内滴注LPS(0.8 ml/kg)染毒;NaHS±+LPS组和NaHS±NS组滴注LPS或NS之前10 min和之后2 h腹腔注射NaHS各0.5 ml(28 μmol/kg).各组取6只大鼠于染毒后12 h制备肺动脉环(PARs),采用离体血管环张力测定技术检测用血红素氧合酶-1(HO-1)抑制剂锌原卟啉Ⅸ(ZnPPⅨ)孵育前后PARs对累积浓度NaHS的舒张反应变化;各组另取6只大鼠于染毒后12 h检测出肺血(EPB)和入肺血(APB)中碳氧血红蛋白(COHb)含量,以其差值反映肺循环CO生成的水平.结果 与对照组相比,滴注LPS后PARs对NaHS的最大舒张反应百分比明显降低[(75.72±7.22)%比(96.40±4.40)%,P＜0.01＝;用ZnPPⅨ孵育PARs后,LPS诱导的PARs对NaHS舒张反应进一步降低[(62.91±8.22)%比(75.72±7.22)%,P＜0.01＝.腹腔注射NaHS可明显逆转LPS诱导的PARs对NaHS的低反应性,PARs对NaHS的最大舒张反应百分比明显升高[(94.65±8.45)%比(75.72±7.22)%,P＜0.01＝;但用ZnPPⅨ孵育PARs后,PARs对NaHS的舒张反应较孵育前显著下降[(83.75±9.76)%比(94.65±8.45)%,P＜0.01＝.NaHS+NS组中PARs对NaHS的舒张反应与对照组相比无明显差异,且在ZnPPⅨ孵育前后也无明显变化.COHb检测结果显示,与对照组相比,滴注LPS后APB和EPB中COHb水平的差值明显增高[(3.12±0.48)%比(2.12±0.32)%,P＜0.05＝;腹腔注射NaHS后,COHb水平的差值[(4.03±0.56)%]较LPS组进一步升高(P＜0.01＝.结论 腹腔注射H2S可以改善LPS诱导的离体肺动脉对H2S的低反应性,其机制可能与增强肺动脉HO-1/CO体系有关.

Abstract：

Objective To explore the effect of hydrogen sulfide (H2S) on abnormal pulmonary artery reactivity induced by lipopolysaccharide (LPS) and its relationship with carbon monoxide (CO). Methods Forty-eight rats were divided into four groups randomly according to table of random number: control group (normal saline, NS), LPS group, a donor of H2S sodium hydrosulfide (NaHS)+LPS group, and NaHS+NS group (n=12 in each group). Rats were given LPS by intratracheal instillation (0. 8 ml/kg). 0. 5 ml of NaHS (28 μmol/kg) was injected intraperitoneally 10 minutes before LPS or NS instillation and 2 hours after LPS or NS instillation in NaHS+LPS and NaHS+NS groups. Twelve hours after instillation of LPS, 6 rats from each group were sacrificed. The pulmonary artery rings (PARs) were prepared and the changes in cumulative relaxation response of PARs to NaHS were detected before and after incubation with an inhibitor of heme oxygenase-1 (HO-1) zinc protoporphyrin Ⅸ (ZnPP Ⅸ ) using isolated vascular ring tension detecting technique. Twelve hours after LPS instillation, the remaining 6 rats in each group were sacrificed, and the contents of carboxyhemoglobin (COHb) in efferent pulmonary blood (EPB) and afferent pulmonary blood (APB) were measured, and the difference between the contents of COHb in EPB and that of APB was calculated to represent content of CO from pulmonary circulation. Results In the present study, compared with control group, after the instillation of LPS the percentage of relaxation response of PARs to NaHS was significantly declined [(75. 72±7. 22)% vs. (96. 40±4. 40)%, P＜0. 01]. After being incubated with ZnPP Ⅸ, the decreased relaxation response of PARs to NaHS induced by LPS was further depressed [(62. 91 ±8. 22) % vs. ( 75. 72 ± 7. 22) %, P ＜ 0. 01]. Administration of NaHS intraperitoneally reversed the hyporesponsiveness of PARs to NaHS, the percentage of relaxation response of PARs to NaHS was significantly increased [(94.65± 8.45)% vs. (75.72 ± 7.22)%, P＜0.01]. However ZnPP Ⅸ also attenuated the effect [(83. 75 ± 9. 76)% vs. (94. 65 ± 8. 45)%, P ＜ 0. 01]. NO significant changes were observed between NaHS+NS group and control group, also between the results before and after ZnPP Ⅸincubation. Compared with control group, the difference between the contents of COHb in EPB and that of APB increased after instillation of LPS [(3. 12±0. 48)% vs. (2. 12±0. 32)%, P＜0. 05], which further increased after intraperitoneal administration of NaHS [(4.03 ± 0. 56) %, P ＜ 0. 01]. Conclusion The results suggested that intraperitoneal administration of H2S could reverse hyporesponsiveness of PARs to H2S induced by LPS, and the result might be related to an intensification of HO-1/CO system in pulmonary artery tissue.     

硫化氢对脂多糖诱导大鼠肺动脉反应性和损伤的影响

目的 观察硫化氢(H2S)对脂多糖(LPS)所致肺动脉反应性紊乱和肺动脉损伤的影响.方法 72只SD大鼠按随机数字表法分为对照组、LPS组、H2S供体硫氢化钠(NaHS)+LPS组和NaHS+生理盐水(NS)组,每组18只.采用气管内滴注0.8 ml/kg LPS(200 μg/200 μl)染毒.滴注LPS之前10 min和之后2 h分别经腹腔注射0.5 ml NaHS(28 μmol/kg).实验12 h处死大鼠,取颈动脉血,检测血清H2S含量;制备肺动脉环(PARs),采用离体血管环张力测定技术检测血管反应性变化;检测肺动脉丙二醛(MDA)含量,并观察肺动脉形态学改变.结果 与对照组相比,滴注LPS后,PARs对苯肾上腺素(PE,10-6 mol/L)的收缩反应(g/mg)明显升高(0.86±0.20比0.56±0.13),对乙酰胆碱(ACh,10-6 mol/L)的舒张反应明显降低[(65.18±7.05)%比(84.13±8.84)%],肺动脉组织MDA含量(mmol/L)升高(32.03±7.81比5.82±0.92),血清中H2S含量(μmol/L)降低(175.23±27.36比238.12±16.38),差异均有统计学意义(P<0.05或P<0.01);组织形态学观察显示,肺动脉内皮细胞和组织结构严重受损.给予NaHS后可明显改善LPS引起的上述变化,血管收缩反应下降[(0.61±0.17) g/mg],血管舒张反应升高[(82.92±9.71)%],肺动脉组织MDA含量下降[(16.88±3.54) mmol/L],血清H2S含量升高[(242.70±38.80) μmol/L],差异有统计学意义(P<0.05或P<0.01);肺动脉内皮细胞和组织结构损伤也得到明显改善.NaHS+NS组除H2S含量显著高于对照组外,余指标与对照组比较差异无统计学意义.结论 外源性应用H2S不仅可逆转LPS引起的肺动脉反应性紊乱,还可减轻LPS引起的肺动脉组织损伤.     

白介素4通过调节巨噬细胞分化促进野百合碱诱导大鼠肺动脉高压模型的肺血管重塑

目的探讨大鼠肺动脉高压模型中白介素-4(IL-4)对巨噬细胞分化和肺血管重塑的影响。方法利用野百合碱(MCT)腹腔注射构建大鼠肺动脉高压模型,以超声心动图及病理改变作为建模成功的观察指标。采用Elisa及荧光定量PCR(qRT-PCR)检测各组血液中IL-4的表达。利用IL-4中和抗体静脉注射肺动脉高压大鼠,抑制体内IL-4的作用,并通过超声心动图及HE染色观察肺动脉高压情况。结果与对照组比较,肺动脉高压组大鼠血液中IL-4的表达(4.01±0.18 vs.9.08±0.25)明显上调(P0.01),大鼠右心室压力(58.00±2.84 vs.20.13±1.89)明显上调(P0.01),且肺血管重塑明显增加。与肺动脉高压组和IgG组比较,IL-4中和抗体组大鼠右心室压力[(42.77±2.04)vs.(58.00±2.84)vs.(57.62±1.44)]明显降低(P0.01),且肺血管重塑明显降低。与肺动脉高压组和IgG组比较,IL-4中和抗体组大鼠肺组织中CD206mRNA表达量[(2.88±0.17)×10-3vs.(4.22±0.08)×10-3vs.(4.36±0.26)×10-3]明显降低(P0.01)。结论肺动脉高压后,上调的IL-4可以促进巨噬细胞向Ⅱ型巨噬细胞分化,进而调节肺动脉的血管重塑过程。     

大鼠左侧颈总动脉与颈外静脉吻合分流性肺动脉高压模型建立

目的：建立大鼠左颈总动脉与左侧颈外静脉吻合分流性肺动脉高压模型。方法成年雄性Wistar大鼠随机分为三组：吻合组（A组,n＝15）,结扎组（B组,n＝15）,正常组（C组,n＝15）。饲养3月后测定血流动力学指标,右心室/左心室＋室间隔质量比,肺组织切片病理变化,ELISA方法测定血浆ET-1浓度,荧光定量PCR测定肺组织表达内皮素-1mRNA的变化。结果与B,C两组相比,A组：右心室收缩压力明显升高,血清内皮素-1水平升高,荧光定量PCR结果显示A组内皮素-1mRNA的相对表达量是B组的2．601倍,是C组的3．373倍,B组是C组的1．297（P＜0．05）。结论通过分离左颈总动脉及左颈外静脉切断后,近心端吻合,可成功建立模拟先天性心脏病的分流性肺动脉高压模型,作为肺动脉高压标志的内皮素-1在此模型中处于高表达状态。     

Drug-associated pulmonary arterial hypertension

Introduction: While pulmonary arterial hypertension remains an uncommon diagnosis, various therapeutic agents are recognized as important associations. These agents are typically categorized into “definite”, “likely”, “possible”, or “unlikely” to cause pulmonary arterial hypertension, based on the strength of evidence.

Objective: This review will focus on those therapeutic agents where there is sufficient literature to adequately comment on the role of the agent in the pathogenesis of pulmonary arterial hypertension.

Methods: A systematic search was conducted using PubMed covering the period September 1970– 2017. The search term utilized was “drug induced pulmonary hypertension”. This resulted in the identification of 853 peer-reviewed articles including case reports. Each paper was then reviewed by the authors for its relevance. The majority of these papers (599) were excluded as they related to systemic hypertension, chronic obstructive pulmonary disease, human immunodeficiency virus, pulmonary fibrosis, alternate differential diagnosis, treatment, basic science, adverse effects of treatment, and pulmonary hypertension secondary to pulmonary embolism.

Agents affecting serotonin metabolism (and related anorexigens): Anorexigens, such as aminorex, fenfluramine, benfluorex, phenylpropanolamine, and dexfenfluramine were the first class of medications recognized to cause pulmonary arterial hypertension. Although most of these medications have now been withdrawn worldwide, they remain important not only from a historical perspective, but because their impact on serotonin metabolism remains relevant. Selective serotonin reuptake inhibitors, tryptophan, and lithium, which affect serotonin metabolism, have also been implicated in the development of pulmonary arterial hypertension.

Interferon and related medications: Interferon alfa and sofosbuvir have been linked to the development of pulmonary arterial hypertension in patients with other risk factors, such as human immunodeficiency virus co-infection.

Antiviral therapies: Sofosbuvir has been associated with two cases of pulmonary artery hypertension in patients with multiple risk factors for its development. Its role in pathogenesis remains unclear.

Small molecule tyrosine kinase inhibitors: Small molecule tyrosine kinase inhibitors represent a relatively new class of medications. Of these dasatinib has the strongest evidence in drug-induced pulmonary arterial hypertension, considered a recognized cause. Nilotinib, ponatinib, carfilzomib, and ruxolitinib are newer agents, which paradoxically have been linked to both cause and treatment for pulmonary arterial hypertension.

Monoclonal antibodies and immune regulating medications: Several case reports have linked some monoclonal antibodies and immune modulating therapies to pulmonary arterial hypertension. There are no large series documenting an increased prevalence of pulmonary arterial hypertension complicating these agents; nonetheless, trastuzumab emtansine, rituximab, bevacizumab, cyclosporine, and leflunomide have all been implicated in case reports.

Opioids and substances of abuse: Buprenorphine and cocaine have been identified as potential causes of pulmonary arterial hypertension. The mechanism by which this occurs is unclear. Tramadol has been demonstrated to cause severe, transient, and reversible pulmonary hypertension.

Chemotherapeutic agents: Alkylating and alkylating-like agents, such as bleomycin, cyclophosphamide, and mitomycin have increased the risk of pulmonary veno-occlusive disease, which may be clinically indistinct from pulmonary arterial hypertension. Thalidomide and paclitaxel have also been implicated as potential causes.

Miscellaneous medications: Protamine appears to be able to cause acute, reversible pulmonary hypertension when bound to heparin. Amiodarone is also capable of causing pulmonary hypertension by way of recognized side effects.

Conclusions: Pulmonary arterial hypertension remains a rare diagnosis, with drug-induced causes even more uncommon, accounting for only 10.5% of cases in large registry series. Despite several agents being implicated in the development of PAH, the supportive evidence is typically limited, based on case series and observational data. Furthermore, even in the drugs with relatively strong associations, factors that predispose an individual to PAH have yet to be elucidated.     

Protective role of protein C inhibitor in monocrotaline-induced pulmonary hypertension

BACKGROUND: Protein C inhibitor (PCI) plays a role in multiple biological processes including fertilization, coagulation, fibrinolysis and kinin systems. OBJECTIVES: We hypothesized that PCI participates in the pathogenesis of pulmonary hypertension. To demonstrate this, we compared the development of pulmonary hypertension in mice overexpressing PCI in the lung with wild-type (WT) mice. Pulmonary hypertension was induced by s.c. injection of 600 mg kg-1 of monocrotaline weekly for 8 weeks. RESULTS: Right ventricular arterial pressure was significantly increased in monocrotaline-treated WT mice compared with that in monocrotaline-treated transgenic mice. Bronchoalveolar lavage fluid (BALF) levels of thrombin-antithrombin complex, monocyte chemoattractant protein-1 and platelet-derived growth factor, and the plasma level of tumor necrosis factor-alpha were significantly increased in monocrotaline-treated WT mice as compared with monocrotaline-treated PCI transgenic mice. Increased level of PCI-thrombin complex was detected in BALF from monocrotaline-treated PCI transgenic mice as compared with saline-treated PCI transgenic mice. CONCLUSIONS: This study showed that increased expression of PCI in the lung is protective against monocrotaline-induced pulmonary hypertension, suggesting a potential beneficial effect of PCI for the therapy of this disease.     

肺动脉高压诊断和治疗进展

慢性阻塞性肺疾病（COPD）引起的肺动脉高压（PAH）是影响患者生存的主要原因。本文重点介绍COPD合并PAH的诊断和治疗进展。     

Computed tomography angiographic parameters of pulmonary artery as prognostic factors of residual pulmonary hypertension after pulmonary endarterectomy

ObjectivesThis study aimed to retrospectively assess using computed tomography pulmonary angiography (CTPA) for predicting residual pulmonary hypertension (RPH) in patients with chronic thromboembolic pulmonary hypertension (CTEPH) after pulmonary endarterectomy (PEA).MethodsWe retrospectively analyzed data of 131 patients with CTEPH who underwent PEA in our center (2008–2015). We measured several diameters of the pulmonary artery and thoracic aorta preoperatively. We evaluated the relationship between these measurements (and their indices) and signs of RPH represented by pulmonary artery systolic pressure (PASP) estimated by echocardiography.ResultsSignificant correlations were observed between the aortopulmonary index and prediction of any residual hypertension and moderate/severe hypertension 1 year after PEA, and any residual hypertension and severe hypertension 2 years after PEA. The aortopulmonary index was significantly related to a reduction in PASP 1 year after the operation. A lower aortopulmonary index (≤0.88 for the ascending aorta and ≤0.64 for the descending aorta) predicted lower RPH.ConclusionsPreoperative CTPA parameters can be used to assess the risk of RPH after PEA. The aortopulmonary index has significant predictive value for RPH and a reduction in PASP after PEA. Lower values of the aortopulmonary index suggest a better outcome after PEA.     

Magnesium aspartate hydrochloride attenuates monocrotaline-induced pulmonary artery hypertension in rats

1. The effect of oral magnesium aspartate hydrochloride on monocrotaline (MCT)-induced pulmonary arterial hypertension was evaluated in rats. 2. A single subcutaneous injection of MCT, a pyrrolizidine alkaloid of plant origin, induces significant morphological changes in pulmonary vessels, pulmonary arterial hypertension and right ventricular hypertrophy in rats by 3 weeks. 3. Two groups of rats (Mg2+ control and Mg2+ + MCT) were started on oral Mg2+ (15.4 g/l magnesium aspartate hydrochloride dissolved in deionized water) 2 weeks before the MCT injection. The rest were given deionized water. At the start of the experiment, the control groups (deionized water and Mg2+) were given normal saline subcutaneously; the other groups (deionized water and Mg2+) were given MCT (60 mg/kg) subcutaneously. 4. Pulmonary artery pressure, right ventricular hypertrophy, lung pathology, organ weights and serum electrolytes were assessed 3 weeks after a single subcutaneous injection of MCT. Seventy-five per cent of the rats treated with MCT and oral Mg2+ (12 out of 16) showed significant reduction in pulmonary arterial hypertension, arterial pathology and right ventricular hypertrophy. 5. Our data indicate that Mg2+ attenuates experimentally induced pulmonary hypertension, possibly either by modulating the intracellular Ca2+ level and/or by directly affecting the pulmonary endothelial cell-smooth muscle cell complex involved in metabolism and maintenance of pulmonary vascular resistance.     

槲皮素对肺动脉平滑肌细胞增殖及大鼠低氧性肺动脉高压的影响

目的研究酪氨酸蛋白激酶抑制剂槲皮素（ｑｕｅｒｃｅｔｉｎ）对血小板衍生生长因子ＰＤＧＦ诱导的平滑肌细胞增殖及低氧性肺动脉高压的影响。方法①用原代培养的小牛肺动脉平滑肌细胞,采用ＭＴＴ比色法、流式细胞及ＷｅｓｔｅｒｎＢｌｏｔ技术对ＰＤＧＦ诱导的肺动脉平滑肌细胞增殖的影响进行分析;②通过大鼠缺氧动物模型观察槲皮素对右心室收缩压（ＲＶＳＰ）,右心室／左心室＋室间隔比值（ＲＶ／ＬＶ＋Ｓ）及肺小动脉图像分析的影响。结果与对照组相比槲皮素处理组能显著地抑制ＰＤＧＦ诱导的肺动脉平滑肌细胞增殖,明显地抑制ＰＤＧＦ诱导的肺动脉平滑肌细胞酪氨酸磷酸化程度,且呈明显剂量依赖性,槲皮素对ＲＶＳＰ、ＲＶ／ＬＶ＋Ｓ及肺小动脉中膜肥厚也有明显的抑制作用。结论槲皮素可显著地抑制ＰＤＧＦ诱导的肺动脉平滑肌细胞增殖,对低氧性肺动脉高压有一定的防治作用。