连翘、麦冬对肺纤维化大鼠TIMP-1表达的影响
|
摘要
目的:观察连翘、麦冬对博来霉素致肺纤维化大鼠肺组织中基质金属蛋白酶9(MMP-9)和基质金属蛋白酶组织抑制剂1(TIMP-1)表达的影响,并初步探讨其抗肺纤维化的作用机制。方法:选取120只Wistar大鼠,随机分为正常对照组、肺纤维化模型组、连翘组、麦冬组和吡非尼酮组。模型组和给药组大鼠的气管内注射博来霉素(5 mg/kg)诱导肺纤维化,正常对照组气管内注射等量生理盐水。造模后第4天,连翘组按生药量1.28 g(kg·d)、麦冬组按生药量1.02 g(kg·d)、吡非尼酮按0.18 g(kg·d)剂量灌胃给药,正常对照组、模型组每日等量灌胃生理盐水,给药体积1 mL/0.1 kg。各组分别在给药后第7天、第14天、第28天随机处死8只大鼠,HE染色观察肺组织的病理变化,酶联免疫法检测大鼠Ⅰ型胶原蛋白水平,免疫组化SP法观察肺组织中MMP-9和TIMP-1表达的变化。结果:与正常对照组比较,模型组大鼠的肺MMP-9和TIMP-1蛋白的表达增加;与模型组比较,连翘组、麦冬组MMP-9蛋白表达减少,连翘组肺组织TIMP-1蛋白表达明显减少,麦冬组第28天肺组织TIMP-1蛋白表达明显减少,但均未恢复正常。结论:连翘、麦冬可抑制肺纤维化大鼠肺组织MMP-9和TIMP-1的合成及分泌,调节两者的比值,这可能是其抑制肺纤维化形成的机制之一。
Objective:To observe the effects of Lianqiao(Forsythia Fructus) and Maidong(Ophiopogon Radix)on the expression of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1) in lung tissue of rats with pulmonary fibrosis induced by bleomycin and to explore its mechanism of treating pulmonary fibrosis. Methods:A total of 120 Wistar rats were randomly divided into normal control group,pulmonary fibrosis model group,Lianqiao group and Maidong group and pirfenidone group. Pulmonary fibrosis was induced by intratracheal injection of bleomycin(5 mg/kg) in the model group and the administration group,and the normal control group was intratracheally injected with the same amount of normal saline. On the 4th day after model establishment,the Lianqiao group was administered with a dosage of 1.28 g(kg·d),the Maidong group with a dosage 1.02 g(kg·d),and pirfenidone was administered at a dosage of 0.18 g(kg·d). After administration,the normal control group and the model group were intragastrically administered with normal saline daily,and the administration volume was 1 mL/0.1 kg. Eight rats were randomly sacrificed on the 7th,14th and 28th day after administration. The pathological changes of lung tissue were observed by HE staining,and the type Ⅰ collagen level was detected by enzyme-linked immunosorbent assay. Immunohistochemistry SP method was used to observe the changes of MMP-9 and TIMP-1 expression in lung tissue. Results:Compared with the normal control group,the expression of MMP-9 and TIMP-1 protein in the lung of the model group increased. Compared with the model group,the expression of MMP-9 protein in the Lianqiao group and Maidong group decreased,the expression of TIMP-1 protein in the Lianqiao group was significantly decreased,and the expression of TIMP-1 protein in lung tissue was significantly decreased on the 28th day of the Maidong group,but none of them returned to normal. Conclusion:Lianqiao and Maidong can inhibit the synthesis and secretion of MMP-9 and TIMP-1 in lung tissue of rats with pulmonary fibrosis,and regulate the ratio of the two,which may be one of the mechanisms to inhibit the formation of pulmonary fibrosis.
Objective:To observe the effects of Lianqiao(Forsythia Fructus) and Maidong(Ophiopogon Radix)on the expression of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1) in lung tissue of rats with pulmonary fibrosis induced by bleomycin and to explore its mechanism of treating pulmonary fibrosis. Methods:A total of 120 Wistar rats were randomly divided into normal control group,pulmonary fibrosis model group,Lianqiao group and Maidong group and pirfenidone group. Pulmonary fibrosis was induced by intratracheal injection of bleomycin(5 mg/kg) in the model group and the administration group,and the normal control group was intratracheally injected with the same amount of normal saline. On the 4th day after model establishment,the Lianqiao group was administered with a dosage of 1.28 g(kg·d),the Maidong group with a dosage 1.02 g(kg·d),and pirfenidone was administered at a dosage of 0.18 g(kg·d). After administration,the normal control group and the model group were intragastrically administered with normal saline daily,and the administration volume was 1 mL/0.1 kg. Eight rats were randomly sacrificed on the 7th,14th and 28th day after administration. The pathological changes of lung tissue were observed by HE staining,and the type Ⅰ collagen level was detected by enzyme-linked immunosorbent assay. Immunohistochemistry SP method was used to observe the changes of MMP-9 and TIMP-1 expression in lung tissue. Results:Compared with the normal control group,the expression of MMP-9 and TIMP-1 protein in the lung of the model group increased. Compared with the model group,the expression of MMP-9 protein in the Lianqiao group and Maidong group decreased,the expression of TIMP-1 protein in the Lianqiao group was significantly decreased,and the expression of TIMP-1 protein in lung tissue was significantly decreased on the 28th day of the Maidong group,but none of them returned to normal. Conclusion:Lianqiao and Maidong can inhibit the synthesis and secretion of MMP-9 and TIMP-1 in lung tissue of rats with pulmonary fibrosis,and regulate the ratio of the two,which may be one of the mechanisms to inhibit the formation of pulmonary fibrosis.
引文
[1]张弦,王强.中医治疗特发性肺纤维化[J].吉林中医药,2014,34(12):1218-1221.
[2]张伟,鲁香凤,张晓梅,等.气管插管快速灌注博莱霉素复制大鼠肺间质纤维化模型[J].中西医结合学报,2008,6(1):60-67
[3]张振龙,蒋春梅.治疗肺纤维化中药复方用药规律文献分析[J].社区医学杂志,2015,13(15):34-36.
[4]黄云鉴,龚婕宁.论肺痹肺痿与肺纤维化的证治规律[J].时珍国医国药,2016,27(6):1439-1441.
[5]李柏颖.清热解毒澄其源——宋康教授治疗特发性肺纤维化经验介绍[J].浙江中西医结合杂志,2008,18(6):359-360.
[6]付义,余晓玲,魏丹霞,等.特发性肺间质纤维化中医治疗探赜[J].中国中医药信息杂志,2016,23(7):108-110.
[7]徐元元,戈伟,徐忠诚.痰热清注射液辅助治疗肺癌急性放射性肺损伤及肺纤维化疗效观察[J].现代中西医结合杂志,2017,26(12):1287-1289,1316.
[8]韩颖萍,黄霞,刘惠霞,等.沙参麦冬汤对肺间质纤维化大鼠氧自由基损伤及细胞外基质代谢的影响[J].中华中医药杂志,2011,26(9):2169-2171.
[8]刘晓艳,陈强.基质金属蛋白酶及其抑制剂在特发性肺纤维化中的作用[J].南昌大学学报(医学版),2015,55(1):88-91,100.
[10]高娟,王添印,韩茹,等. MMP-9、TIMP-1在肺纤维化中作用的研究进展[J].山东医药,2017,57(26):104-106.
[11]唐学义,王思勤,郑素歌,等. IPF患者基质金属蛋白酶及其抑制剂检测的临床意义[J].医药论坛杂志,2010,31(11):59-61.
[12]金晓光,代华平,庞宝森,等.博来霉素致大鼠肺纤维化模型肺组织的动态病理变化及其发生机制[J].中国病理生理杂志,2009,25(4):708-714.
[13]范碧君,蒋捍东. 2015版特发性肺纤维化治疗指南解读[J].世界临床药物,2016,37(7):453-456.
[2]张伟,鲁香凤,张晓梅,等.气管插管快速灌注博莱霉素复制大鼠肺间质纤维化模型[J].中西医结合学报,2008,6(1):60-67
[3]张振龙,蒋春梅.治疗肺纤维化中药复方用药规律文献分析[J].社区医学杂志,2015,13(15):34-36.
[4]黄云鉴,龚婕宁.论肺痹肺痿与肺纤维化的证治规律[J].时珍国医国药,2016,27(6):1439-1441.
[5]李柏颖.清热解毒澄其源——宋康教授治疗特发性肺纤维化经验介绍[J].浙江中西医结合杂志,2008,18(6):359-360.
[6]付义,余晓玲,魏丹霞,等.特发性肺间质纤维化中医治疗探赜[J].中国中医药信息杂志,2016,23(7):108-110.
[7]徐元元,戈伟,徐忠诚.痰热清注射液辅助治疗肺癌急性放射性肺损伤及肺纤维化疗效观察[J].现代中西医结合杂志,2017,26(12):1287-1289,1316.
[8]韩颖萍,黄霞,刘惠霞,等.沙参麦冬汤对肺间质纤维化大鼠氧自由基损伤及细胞外基质代谢的影响[J].中华中医药杂志,2011,26(9):2169-2171.
[8]刘晓艳,陈强.基质金属蛋白酶及其抑制剂在特发性肺纤维化中的作用[J].南昌大学学报(医学版),2015,55(1):88-91,100.
[10]高娟,王添印,韩茹,等. MMP-9、TIMP-1在肺纤维化中作用的研究进展[J].山东医药,2017,57(26):104-106.
[11]唐学义,王思勤,郑素歌,等. IPF患者基质金属蛋白酶及其抑制剂检测的临床意义[J].医药论坛杂志,2010,31(11):59-61.
[12]金晓光,代华平,庞宝森,等.博来霉素致大鼠肺纤维化模型肺组织的动态病理变化及其发生机制[J].中国病理生理杂志,2009,25(4):708-714.
[13]范碧君,蒋捍东. 2015版特发性肺纤维化治疗指南解读[J].世界临床药物,2016,37(7):453-456.