藻蓝蛋白对野百合碱诱导肺动脉高压大鼠肺血管重构的影响
Effect of phycocyanin on pulmonary vascular remodeling in rats with monocrotaline-induced pulmonary arterial hypertension
赵水炎 1高帅 1曹越 1唐雅琪 1泮思林1
作者信息
- 1. 青岛大学附属妇女儿童医院心脏中心,青岛 266034
- 折叠
摘要
目的 探讨藻蓝蛋白改善肺动脉高压(PAH)大鼠肺血管重构的机制.方法 将30只雄性Sprague-Dawley(SD)大鼠,采用随机数表法分为3组:正常对照组(对照组)、PAH模型组(PAH组)、藻蓝蛋白治疗组(治疗组),每组10只.腹腔注射野百合碱(MCT)建立PAH模型,治疗组在造模后第15天给予藻蓝蛋白灌胃,持续14d.测量右心室收缩压(RVSP),计算右心室肥厚指数(RVHI).苏木精-伊红(HE)染色、马松(Masson)染色观察肺血管,蛋白印迹法(Western blot)检测转化生长因子-β1(TGF-β1)、核转录因子-κB(NF-κB)、B淋巴细胞瘤-2(bcl-2)、bcl-2相关X蛋白(bax)、天冬氨酸特异半胱氨酸蛋白酶-3(Caspase-3)、α-肌动蛋白(α-SMA)、增殖细胞核抗原(PCNA)表达,免疫荧光定位α-SMA和PCNA.多组比较采用单因素方差分析.结果 治疗组RVSP、RVHI 低于 PAH 组[(24.60±3.33)mmHg 比(40.49±3.76)mmHg,F=140.998,P＜0.01;(32.23±2.99)％比(43.45±3.10)％,F=90.774,P＜0.01],肺小动脉中膜厚度及中膜面积百分比低于 PAH 组(0.579±0.013 比 0.736±0.023,F=251.494,P＜0.01;0.574±0.063 比 0.917± 0.036,F=84.346,P＜0.05,肺血管胶原沉积低于 PAH 组[(9.81±0.66)％比(23.23±2.05)％,F=294.181,P＜0.01].治疗组 TGF-β1、bcl-2、α-SMA 和 PCNA 相对表达量、磷酸化 NF-κB(p-NF-κB)/NF-κB 低于 PAH 组(0.813±0.077 比 1.097±0.056,F=75.485,P＜0.01;0.902± 0.030 比 1.231±0.123,F=52.224,P＜0.01;1.089±0.044 比 1.283±0.049,F=85.388,P＜0.01;1.218±0.008 比 1.673±0.139,F=33.825,P＜0.05;0.463±0.025 比 0.875±0.013,F=447.19,P＜0.01);bax、Caspase-3 相对表达量高于 PAH 组(1.188±0.072 比 0.738±0.048,F=63.635,P＜0.01;1.750±0.028 比 1.432±0.017,F=31.684,P＜0.01).治疗组 α-SMA、PCNA 荧光强度低于对照组.结论 藻蓝蛋白可通过抑制肺血管胶原沉积,减少炎性因子表达,抑制细胞增殖,促进凋亡,减轻大鼠PAH肺血管重构.
Abstract
Objective To investigate the mechanism of phycocyanin improving pulmonary vascular remodeling in rats with pulmonary arterial hypertension(PAH).Methods We randomly divided 30 male Sprague-Dawley(SD)rats into 3 groups:normal control group,PAH group,and phycocyanin treatment group,with 10 rats in each group.The PAH model was induced by intraperitoneal injection of monocrotal-ine(MCT).The treatment group was given phycocyanin by gavage on the 15 th day.Effects on right ven-tricular systolic blood pressure(RVSP)and right ventricular hypertrophy index(RVHI)were examined.The changes of pulmonary vessels were examined by hematoxylin-eosin(HE)and Masson staining.The ex-pression of transforming growth factor-β1(TGF-β1),nuclear factor kappa-B(NF-κB),B-cell lymphoma-2(bcl-2),cysteinyl aspartate-specific protease(Caspase)-3,α-smooth muscle actin(α-SMA)and prolifera-ting cell nuclear antigen(PCNA)was detected by Western blotting.The expression of α-SMA and PCNA was evaluated by immunofluorescence.Statistical significance was determined by one-way ANOVA.Results RVSP and RVHI in the treatment group were lower than in the PAH group[(24.60± 3.33)mmHg vs.(40.49±3.76)mmHg,F=140.998,P＜0.01;(32.23±2.99)％vs.(43.45± 3.10)％,F=90.774,P＜0.01].The percentage of pulmonary arteriole media thickness and media area decreased in the treatment group and pulmonary vascular collagen deposition decreased after treatment(0.579±0.013 vs.0.736±0.023,F=251.494,P＜0.01;0.574±0.063 vs.0.917±0.036,F=84.346,P＜0.05;(9.81±0.66)％vs.(23.23±2.05)％,F=294.181,P＜0.01).The expression levels of TGF-β1,bcl-2,α-SMA,PCNA and phosphorylated NF-κB(p-NF-κB)/NF-κB were suppressed whereas bcl-2 associated X protein(bax)and Caspase-3 were elevated in the treatment group(0.813±0.077 vs.1.097±0.056,F=75.485,P＜0.01;0.902±0.030 vs.1.231±0.123,F=52.224,P＜0.01;1.089±0.044 vs.1.283±0.049,F=85.388,P＜0.01;1.218±0.008 vs.1.673±0.139,F=33.825,P＜0.05;0.463±0.025 vs.0.875±0.013,F=447.19,P＜0.01;1.188±0.072 vs.0.738±0.048,F=63.635,P＜0.01;1.750±0.028 vs.1.432±0.017,F=31.684,P＜0.01).The fluorescence inten-sity of α-SMA and PCNA in the treatment group was lower than in the control group.Conclusion Phyco-cyanin alleviates pulmonary vascular remodeling of PAH in rats by inhibiting pulmonary vascular collagen deposition,reducing inflammatory factor expression,inhibiting cell proliferation and promoting apoptosis.
关键词
藻蓝蛋白/肺动脉高压/肺血管重构/炎性反应/细胞增殖Key words
Phycocyanin/Pulmonary arterial hypertension/Pulmonary vascular remodeling/Inflammatory response/Cell proliferation引用本文复制引用
基金项目
国家自然科学基金(82271725)
国家自然科学基金(81970249)
山东省泰山学者工程资助(2018)()
青岛市科技惠民示范引导专项(23-2-8-smjk-10-nsh)
出版年
2024
NETL
NSTL
万方数据