摘要
目的
观察丰白散对慢性阻塞性肺疾病模型大鼠行为学的改变,以及肺组织中细胞凋亡的变化,肺组织匀浆中VEGF及其受体2表达的影响,初步探讨丰白散对COPD大鼠模型的治疗作用及其可能的作用机制。
方法
选用健康Wistar大鼠(雄性)46只,6月龄,体重260～350g,随机分为两组,正常组10只,造模组36只(模型组、中药组、西药组);(1)正常组:不予熏烟,自由喂养。(2)造模组:第1、14天气管内注入脂多糖(LPS)各200ug/200ul,第2-13天、15-28天每日上、下午在密闭箱内熏烟0.5h。从实验第1天开始用生理盐水1 mL/只·天灌胃,连续28天。(3)丰白散组(以下简称中药组):造模方法同模型组,从实验第1天开始用中药煎剂灌胃1 mL/只·天,连续28天。(4)西药组:造模方法同模型组,从实验第一天开始用西药醋酸泼尼松混悬液1 mL/只·天灌胃,连续28天。然后模型组大鼠肺组织切片HE染色,光学显微镜下观察肺组织结构的病理形态学变化;TUNEL检测细胞凋亡;逆转录聚合酶链反应(RT-PCR)半定量分析比较各组大鼠肺组织VEGF、KDR mRNA基因表达水平。
结果
1).COPD模型组大鼠有慢性支气管炎、阻塞性肺气肿的特征性病理改变。同时进行中药组和西药组干预治疗之后,上述病理变化相对减轻。
2).模型组肺泡间隔细胞AI高于正常组、中药组,西药组,差异有统计学意义(P＜0.05),与正常组比较,模型组、中药组、西药组肺泡间隔细胞AI,差异有统计学意义。
3).从RT-PCR结果观察,基础状态下肺内存在VEGF和KDRmRNA转录表达。与模型组相比较,正常组、中药组和西药组VEGFmRNA和KDRmRNA表达,差异具有统计学意义(P＜0.05);与正常组比较,模型组、中药组和西药组VEGFmRNA和KDRmRNA表达,差异亦有统计学意义(P＜0.05)。
结论
1).本实验采用烟雾暴露加在气管内滴入脂多糖制作COPD大鼠模型,大鼠出现COPD的一些临床症状,模型组HE染色出现慢性支气管炎和阻塞性肺气肿典型的病理特征,说明该模型的建立是成功的。
2).丰白散能增强COPD模型大鼠VEGF及其受体KDR的表达,减少肺泡间隔细胞的凋亡,机理可能是该复方通过影响VEGF及其受体2的表达延缓细胞凋亡,同时推测可能是丰白散治疗COPD的部分作用机制。
Objectives
To observe the effect of Feng Baisan on ethology,the chang of apoptosis and the expression of VEGF and KDR in lung tissue of the rats with chronic obstructive pulmonary disease,and explore the therapeutical effect on COPD rats and the potential mechanism of action of Feng Baisan.
Methods
46 Wistar healthy male rats of six months old,weighted 260-350g, were randomly divided into four groups:a normal group(n=10,perfused smoking,flee drinking water );a COPD model group、a Fengbaisan group(that was traditional Chinese medicine group,TCM group),a western medicine group,each 12;a COPD model group which was established by intratracheal instillation of 200 ug lipopolysaccharide once for every two weeks(twice),and exposed to 5%cigarete smoke for 0.5h twice per day for 4 weeks;a TCM group which COPD group plus the traditional Chinese medicine fengbaisan treatment group;a western medicine group which COPD group plus western medicine treatment group。Lung tissue sections stained by HE were observed to study the morphological alternation;terminal deoxynucleotide transferase-mediated dUTP nick-end labeling(TUNEL) detection of apoptosis;The expressions of VEGFmRNA and KDRmRNA were determined by reverse transcription-polymerase chain reaction(RT-PCR)。
Results
1).There were specific pathognomonic features of chronic bronchitis and obstructive emphysema in COPD rat model group,At the same time,TCM and western medicine treatment group after the intervention, relative to mitigate the above-mentioned pathological changes。
2).The number of TUNEL+ cells in alveolar septa was significantly increased in model group as compared with normal group and TCM group、western medicine group interfered group(P＜0.05),The number of TUNEL+ cells in alveolar septa was significantly deseased in normal group as compared with model group and TCM group、western medi- cine group interfered group(P＜0.05)。
3).The observation of results from RT-PCR,based on the existence of a state of expression of VEGF and KDRmRNA in lung.Compared with model group,the expression of VEGFmRNA and KDR mRNA of normal group,TCM and western medicine group,the differ-ence was significant(P＜0.05);with the normal group,the expression of VEGFmRNA and KDRmRNA of model group,TCM and western medicine group,the difference was still significant(P＜0.05).
Conclusions
1).The presentation of results of the study that the model was established by smoking and injected lipopolysaccharide in gas was blest, because the model rats had the changes of praxiology and pathology.
2).Fengbaisan can raise the expressions of VEGFmRNA and KDRmRNA,then prevent apoptosis,which may be part of the effective mechanism of treatment of COPD model in rats.
引文
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