氨基胍治疗大鼠肺纤维化的效果及相关机制研究

Researches on the Effect of Aminoguanidin on Pulmonary Fibrosis in Rats and Its Related Mechanism

为探讨氨基胍对肺纤维化大鼠肺的影响及机制,本研究选取健康清洁级SD大鼠45只作为研究对象。雌雄各半,随机分为对照组、模型组和观察组,每组15只。模型组和观察组大鼠采用博莱霉素建立肺纤维化模型,其中模型组每天腹腔注射1 mL/kg生理盐水,观察组每天腹腔注射20 mg/kg氨基胍,对照组每天腹腔注射1 mL/kg生理盐水。14 d后,采用明胶酶谱法检测基质金属蛋白酶-9 (MMP-9)和MMP-13活性,HE染色观察肺组织,天狼猩红染色观察肺间质胶原以及免疫组化染色观察结缔组织生长因子(CTGF)表达。通过实验发现,观察组活性MMP-9含量总积分密度值(IDV)为4.200±1.304,明显低于模型组(p<0.05),与对照组比较差异无统计学意义(p>0.05);观察组和模型组活性MMP-13含量IDV比较差异无统计学意义(p>0.05),均明显高于对照组(p<0.05);观察组CTGF表达积分光密度值(IOD)为13.281±2.015,明显低于模型组(p<0.05),但仍高于对照组(p<0.05);HE染色观察,相比较模型组,观察组肺纤维化表现有所减轻;观察组间质胶原面积比例为(0.430±0.120)%,明显低于模型组(p<0.05),与对照组比较差异无统计学意义(p>0.05)。本研究表明氨基胍能有效治疗大鼠肺纤维化,其机制可能与抑制MMP-9活性和CTGF表达有关。

In order to investigate the effect and mechanism of Aminoguanidin(AG)on lung in rats with pulmonary fibrosis,45 healthy and clean SD rats were selected as the research object.Male and female rats were randomly divided into control group,model group and observation group,with each group of 15 rats.The model group and the observation group were applied with bleomycin to establish a pulmonary fibrosis model.The model group was intraperitoneally injected with 1 mL/kg normal saline every day.The observation group was intraperitoneally injected with 20 mg/kg Aminoguanidin every day.The control group was intraperitoneally injected with 1 mL/kg normal saline everyday.After 14 d,the activities of matrix metalloproteinase-9(MMP-9)and MMP-13 were detected by gelatin zymography.The lung tissues were observed by HE staining.The interstitial collagen was observed by Sirius red staining,and immunohistochemical staining was applied to observe the expression of connective tissue growth factor(CTGF).Through experiments,the total integrated densityvalue(IDV)of the active MMP-9 content in the observation group was 4.200±1.304,which was significantly lower than that in the model group(p<0.05),and there was no statistically significant difference compared with the control group(p>0.05).There was no statistically significant difference in theIDV of active MMP-13 between the observation group and the model group(p>0.05),which was significantly higher than that of the control group(p<0.05).The integrated optical density(IOD)of CTGF expression in the observation group was 13.281±2.015,which was significantly lower than that in the model group(p<0.05),but still higher than that in the control group(p<0.05).In the HE staining observation,compared with the model group,the pulmonary fibrosis of the observation group was reduced.The ratio of interstitial collagen area in the observation group was(0.430±0.120)%,Which was significantly lower than that in the model group(p<0.05).And there was no statistically significant difference compared with the control group(p>0.05).This study demonstrated that Aminoguanidin could effectively treat pulmonary fibrosis in rats,and its mechanism might be related to inhibition of MMP-9 activityand CTGF expression.