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大黄对新生大鼠支气管肺发育不良的保护作用

Protective effect of Rhubarb on newborn rats with bronchopulmonary dysplasia

摘要:

目的 探讨大黄对支气管肺发育不良(BPD)的影响及其可能的作用机制.方法 将出生4d的SD大鼠按随机数字表法随机分为4组(每组16只):空气+9 g/L盐水组、空气+大黄组、高氧+9 g/L盐水组、高氧+大黄组.前2组暴露于空气中,后2组暴露于600 mL/L氧中.大黄干预组(空气+大黄组、高氧+大黄组)大鼠每天予大黄600 mg/kg灌胃,对照组(空气+9 g/L盐水组、高氧+9 g/L盐水组)大鼠给予等量9 g/L盐水灌胃.各组新生鼠于出生后14、21 d按随机数字表法随机抽取8只,麻醉后处死并取肺组织.苏木精-伊红(HE)染色检测肺组织形态学改变,原位末端标记法(TUNEL)检测肺组织细胞凋亡,Western blot检测Fas蛋白表达,分光光度计检测Caspase-8和Caspase-3酶活性.结果 与空气+9 g/L盐水组比较,高氧+9 g/L盐水组于大鼠出生后14、21 d时均可见明显的肺组织病理改变(肺泡数目减少,体积增大,肺结构简单化),肺组织细胞凋亡指数(39.91±1.91比10.11±1.64,48.80 ±4.51比12.90±3.18)、Fas蛋白表达量(0.47 ±0.02比0.21±0.01,0.55±0.02比0.22±0.01)、Caspase-8(52.59±1.23比40.74±1.08,60.20±3.48比40.39±2.47)和Caspase-3酶活性(57.17±1.88比42.00±1.19,64.57 ±2.79比41.54±1.27)均显著增加,差异均有统计学意义(均P <0.001).高氧+大黄组与相应时间点的高氧+9 g/L盐水组比较,肺组织病理损伤明显改善,肺组织凋亡细胞减少(26.49±2.65比22.97±3.66),Fas蛋白表达量降低(0.27±0.03比0.31±0.01),Caspase-8(32.70±2.69比30.66±4.48)和Caspase-3(44.94±1.60比44.59±1.66)酶活性均明显降低,差异均有统计学意义(均P <0.001).结论 大黄可通过减少新生大鼠肺组织细胞凋亡减轻BPD的高氧肺损伤,其机制可能涉及Fas凋亡途径.

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abstracts:

Objective To investigate the effect of Rhubarb on bronchopulmonary dysplasia(BPD) and its potential mechanism.Methods Neonatal Sprague-Dawley rats (postnatal day 4) were exposed to 600 mL/L 02 to induce BPD.The experimental rats were randomly divided into 4 groups with 16 in each group:air + saline group,air +Rhubarb (600 mg/kg)group,and hyperoxia + saline group,and hyperoxia + Rhubarb group.The rats were sacrificed and lung tissues were obtained on day 14 and 21 after birth.Hematoxylin-easin staining was used to detect the pathomorphology of the lungs.Apoptosis of the lung tissue was detected by means of TdT-mediated dUTP nick-end labeling (TUNEL).The expression of Fas was detected by adopting Western blot.The activity of Caspase-8 and Caspase-3was detected by using spectrophotometer.Results The lung structure of rats was markedly abnormal (decreased,enlarged and simplified alveoli) after being exposed to hyperoxia at any time point.The apoptosis indexes (39.91 ± 1.91vs.10.11 ± 1.64,48.80 ± 4.51 vs.12.90 ± 3.18),the expression levels of Fas (0.47 ± 0.02 vs.0.21 ± 0.01,0.55 ±0.02 vs.0.22 ±0.01) and the activities of Caspase-8 (52.59 ± 1.23 vs.40.74 ± 1.08,60.20 ± 3.48 vs.40.39 ±2.47) and Caspsase 3 (57.17 ± 1.88 vs.42.00 ± 1.19,64.57 ± 2.79 vs.41.54 ± 1.27) were all increased in the hyperoxia + saline group compared with those in the air + saline group on day 14 and 21,and the differences were statistically significant(all P < 0.001).However,compared with the hyperoxia + saline group,Rhubarb administration dramatically decreased pulmonary apoptosis index (26.49 ± 2.65 vs.22.97 ± 3.66),Fas protein level (0.27 ± 0.03 vs.0.31 ± 0.01) and the activity of Caspase-8 (32.70 ± 2.69 vs.30.66 ± 4.48) and Caspase-3 (44.94 ± 1.60 vs.44.59 ± 1.66),and the differences were statistically significant (all P < 0.001).Meanwhile,lung damage after hyperoxia was significantly attenuated in the hyperoxia + Rhubarb group.Conclusion Rhubarb can reduce the hyperoxic lung injury of BPD by reducing the apoptosis of newborn rat lung tissue cells,and the mechanism may involve the apoptosis pathway of Fas.

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作者: 尹玲玲 [1] 唐丽君 [1] 黄为民 [1]
栏目名称: 实验研究
DOI: 10.3760/cma.j.issn.2095-428X.2019.01.014
发布时间: 2019-05-29
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