虎杖对肺纤维化大鼠肺泡灌洗液TNF-α和PDGF影响的研究(1)
摘 要:目的:动态观察虎杖对肺纤维化大鼠肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)和血小板衍化生长因子(PDGF)两种细胞因子表达及肺组织病理变化的影响。方法:雄性SD大鼠,随机分成4组:正常对照组、模型组、地塞米松组、虎杖预防组。除正常对照组外,其余3组均采用博莱霉素(BLM)制作肺纤维化大鼠模型,每日灌胃给药,造模后3、7、14、28、42天取材。采用酶联免疫吸附(ELISA)法检测BALF中TNF-α和PDGF的含量,采用光学显微镜观察肺组织病理变化。结果:与模型组相比,地塞米松、虎杖预防组肺泡炎与肺纤维化病理程度显著降低;与模型组比较,地塞米松、虎杖预防组BALF中TNF-α和PDGF水平在早期明显下降。结论:虎杖能够降低肺组织肺泡炎和肺纤维化病理程度,下调BALF中TNF-α和PDGF水平,从而抑制或延缓博莱霉素致大鼠肺纤维化的发生和发展。
关键词:肺纤维化;BALF;TNF-α;PDGF;虎杖
中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)01-0043-05
Study on the Effects of Polygonum Cuspidate on TNF-α and PDGF in BALF of Pulmonary Fibrosis Rats
XIA Yongliang,LUO Xianfang,SONG Kang, WANG Yuan
(The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310006, Zhejiang, China)
Abstract:Objective:To observe dynamically the effects of polygonum cuspidate on the expression of tumor necrosis factor-α (TNF-α) and platelet-derived growth factor (PDGF) in the bronchoalveolar lavage fluid (BALF) of pulmonary fibrosis rats and the pathological changes of lung tissue.Methods:100 male SD rats were randomly divided into 4 groups:the normal control group (n=25), the model group (n=25), the dexamethasone (DXM) group (n=25), the polygonum cuspidate prophylactic group (n=25).Except the normal control group, other 3 groups were all adopted bleomycin(BLM) to set up rat model with pulmonary fibrosis by intragastric administration every day, and the materials were draw 3, 7, 14, 28,42 days after making models. The contents of TNF-α and PDGF in BALF were detected with Enzyme-linked Immunosorbent Assay (ELISA), and the pathological changes of lung tissue were observed with optical microscope.Results: Compared with the model group, the degrees of alveolitis and pulmonary fibrosis decreased significantly in the dexamethasone group and the polygonum cuspidate prophylactic group.Compared with the model group, the levels of TNF-α and PDGF in BALF decreased significantly in dexamethasone group and the polygonum cuspidate prophylactic group.Conclusion: Polygonum cuspidate can decrease the degree of alveolitis and pulmonary fibrosis decreased significantly in the dexamethasone group and the polygonum cuspidate prophylactic group, as well as TNF-α and PDGF in BALF, so as to inhibit and delay the occurrence and development of pulmonary fibrosis of rats induced by bleomycin(BLM).
Key words:pulmonary fibrosis; BALF; TNF-α; PDGF; polygonum cuspidate
肺纤维化是多种肺系疾病或药物、放射线等损害发展到晚期的一种常见病理变化和共同结局,其患病率和病死率在呼吸系统疾病中日趋增长,但临床尚缺少有效的治疗方法和手段。本实验通过虎杖对肺纤维化大鼠进行干预,观察肺泡灌洗液中TNF-α和PDGF的蛋白的表达及肺组织的病理变化,探讨虎杖对这两种细胞因子的影响及对肺纤维化防治作用。
1材料和方法
1.1实验材料
1.1.1实验动物雄性SD大鼠145只,清洁级,体重(200±10)g;购自上海实验动物中心,由浙江中医药大学动物实验中心饲养(环境温度25℃,湿度70%)。
1.1.2实验用药虎杖流浸膏(浓度1g/mL),由浙江省中医院制剂室提供;地塞米松,5mg/支,浙江仙居制药股份有限公司生产,批号:070509。
1.1.3 主要试剂和仪器大鼠TNF-α ELISA试剂盒(晶美生物工程有限公司),大鼠PDGF ELISA试剂盒(UNIONHONESE&ADL公司),Bio-Rad Model 550酶标测试仪等。
1.2实验方法
1.2.1动物造模主要参考文献报道[1]。
1.2.2分组及给药SD雄性大鼠100只,随机分成4组:①正常对照组(25只),②模型组(25只),③地塞米松组(25只),④虎杖预防组(25只)。正常对照组和模型组于造模后当天开始每天灌胃生理盐水4mL/kg,地塞米松组于造模后当天开始每天灌胃地塞米松4mL/kg(即1mg/kg),虎杖预防组于造模前2天开始每天灌胃中药虎杖4mL/kg(相当于生药量1g/mL)。每周称重,正常对照组、模型组、地塞米松组及虎杖预防组分别于造模后3、7、14、28、42天各处死5只。 1.2.3标本收集, 百拇医药(夏永良 骆仙芳 宋 康 王 媛)
关键词:肺纤维化;BALF;TNF-α;PDGF;虎杖
中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)01-0043-05
Study on the Effects of Polygonum Cuspidate on TNF-α and PDGF in BALF of Pulmonary Fibrosis Rats
XIA Yongliang,LUO Xianfang,SONG Kang, WANG Yuan
(The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou 310006, Zhejiang, China)
Abstract:Objective:To observe dynamically the effects of polygonum cuspidate on the expression of tumor necrosis factor-α (TNF-α) and platelet-derived growth factor (PDGF) in the bronchoalveolar lavage fluid (BALF) of pulmonary fibrosis rats and the pathological changes of lung tissue.Methods:100 male SD rats were randomly divided into 4 groups:the normal control group (n=25), the model group (n=25), the dexamethasone (DXM) group (n=25), the polygonum cuspidate prophylactic group (n=25).Except the normal control group, other 3 groups were all adopted bleomycin(BLM) to set up rat model with pulmonary fibrosis by intragastric administration every day, and the materials were draw 3, 7, 14, 28,42 days after making models. The contents of TNF-α and PDGF in BALF were detected with Enzyme-linked Immunosorbent Assay (ELISA), and the pathological changes of lung tissue were observed with optical microscope.Results: Compared with the model group, the degrees of alveolitis and pulmonary fibrosis decreased significantly in the dexamethasone group and the polygonum cuspidate prophylactic group.Compared with the model group, the levels of TNF-α and PDGF in BALF decreased significantly in dexamethasone group and the polygonum cuspidate prophylactic group.Conclusion: Polygonum cuspidate can decrease the degree of alveolitis and pulmonary fibrosis decreased significantly in the dexamethasone group and the polygonum cuspidate prophylactic group, as well as TNF-α and PDGF in BALF, so as to inhibit and delay the occurrence and development of pulmonary fibrosis of rats induced by bleomycin(BLM).
Key words:pulmonary fibrosis; BALF; TNF-α; PDGF; polygonum cuspidate
肺纤维化是多种肺系疾病或药物、放射线等损害发展到晚期的一种常见病理变化和共同结局,其患病率和病死率在呼吸系统疾病中日趋增长,但临床尚缺少有效的治疗方法和手段。本实验通过虎杖对肺纤维化大鼠进行干预,观察肺泡灌洗液中TNF-α和PDGF的蛋白的表达及肺组织的病理变化,探讨虎杖对这两种细胞因子的影响及对肺纤维化防治作用。
1材料和方法
1.1实验材料
1.1.1实验动物雄性SD大鼠145只,清洁级,体重(200±10)g;购自上海实验动物中心,由浙江中医药大学动物实验中心饲养(环境温度25℃,湿度70%)。
1.1.2实验用药虎杖流浸膏(浓度1g/mL),由浙江省中医院制剂室提供;地塞米松,5mg/支,浙江仙居制药股份有限公司生产,批号:070509。
1.1.3 主要试剂和仪器大鼠TNF-α ELISA试剂盒(晶美生物工程有限公司),大鼠PDGF ELISA试剂盒(UNIONHONESE&ADL公司),Bio-Rad Model 550酶标测试仪等。
1.2实验方法
1.2.1动物造模主要参考文献报道[1]。
1.2.2分组及给药SD雄性大鼠100只,随机分成4组:①正常对照组(25只),②模型组(25只),③地塞米松组(25只),④虎杖预防组(25只)。正常对照组和模型组于造模后当天开始每天灌胃生理盐水4mL/kg,地塞米松组于造模后当天开始每天灌胃地塞米松4mL/kg(即1mg/kg),虎杖预防组于造模前2天开始每天灌胃中药虎杖4mL/kg(相当于生药量1g/mL)。每周称重,正常对照组、模型组、地塞米松组及虎杖预防组分别于造模后3、7、14、28、42天各处死5只。 1.2.3标本收集, 百拇医药(夏永良 骆仙芳 宋 康 王 媛)