活血定喘汤对哮喘小鼠气道重塑的实验研究
摘要
目的：通过观察Balb/c小鼠血浆MMP-9与TIMP-1的相对含量及二者比值变化及血浆TGF-β1的含量和肺组织形态学的变化,探讨活血定喘汤(祛痰化瘀中药)对哮喘小鼠MMP-9/TIMP-1平衡和气道重塑的影响及其作用机理。
方法：健康Balb/c小鼠70只随机分为正常组、模型组、西药组、活血定喘汤低剂量组、高剂量组。以卵白蛋白致敏激发小鼠制作哮喘气道重塑模型,正常组、模型组以生理盐水灌胃,其余三组分别给予相应的药物干预。实验结束后采用ELISA法检测血浆MMP-9与TIMP-1及TGF-β1,光镜下观察肺脏病理组织形态。
结果：肺组织病理切片显示,活血定喘汤能显著减少炎性细胞的浸润,肺泡壁水肿及支气管平滑肌的增厚,能抑制哮喘气道重塑的发生；血浆检测显示模型组及治疗各组与正常组相比MMP-9与TIMP-1的水平均明显升高,经治疗后均有降低,其中西药组和活血定喘汤高剂量组与模型组之间有明显差异(P<0.01)；同时除正常组之外,其余各组都存在着MMP-9/TIMP-1比例的失衡,模型组MMP-9/TIMP-1的比值出现倒置失衡；血浆TGF-β1的检测结果显示：与正常组比较模型组中TGF-β1的含量显著升高,两组之间存在明显差异(P<0.01),经治疗后高剂量组及西药组上述指标均显著降低,与模型组之间有明显差异(P<0.01),而高剂量组及西药组作用之间无明显差异。
结论：活血定喘汤能抑制哮喘气道重塑的进程,其部分机制可能通过调节哮喘气道重塑小鼠血浆MMP-9、TIMP-1、TGF-β1的水平及MMP-9/TIMP-1之间的平衡实现的。
Objective:Observed the relative content of plasma MMP-9 and TIMP-1, the change of plasma TGF-β1 and lung morphology in Balb/c mice, so as to explore the effect and mechanism of Huoxue Dingchuan Soup(HDS) to MMP-9/TIMP-1 balance and asthmatic airway remodeling.
Methods:Balb/c mice 70 were randomly divided into normal group, model group, western medicine group, HDS low-dose group and high-dose group. Asthmatic airway remodeling model was established by repeated inhalation of ovalbumin. The normal group and model group administrated water, the other groups respectively administrated the corresponding medicines. At the end, determined plasma MMP-9,TIMP-1 and TGFβ1 by ELISA, observed the pathomorphology of lung under light microscope.
Results::The pathological changes in lung tissue showed that HDS can significantly reduce the infiltration of inflammatory cells, alveolar wall edema and thickening of bronchial smooth muscle, can inhibit the occurrence of airway remodeling; model group compared with the control group, the level of MMP-9 and TIMP-1 except the normal group were significantly increased, after treatment they were significantly decreased, in which western medicine group and HDS high-dose group have significant difference comparing with the model group (P<0.01); In addition to the normal group at the same time, the other groups of MMP-9/TIMP-1 are imbalance in the ratio, the model group of TIMP-1 over-proportional increase eventually lead to an imbalance; plasma TGF-β1 results showed that compared with the normal group, TGF-β1 levels of model group were significantly increased, there is significant difference between the two groups (P<0.01), after treatment, the above indices of high-dose group and western medicine group decreased significantly and were significant differences compared with the model (P <0.01), while the high-dose group and western medicine group were no significant difference.
Conclusion:HDS might suppress asthmatic airway remodeling by regulating the level of MMP-9, TIMP-1, TGF-β1 in the lung tissue of asthmatic airway remodeling mice and MMP-9/TIMP-1 balance.
方法：健康Balb/c小鼠70只随机分为正常组、模型组、西药组、活血定喘汤低剂量组、高剂量组。以卵白蛋白致敏激发小鼠制作哮喘气道重塑模型,正常组、模型组以生理盐水灌胃,其余三组分别给予相应的药物干预。实验结束后采用ELISA法检测血浆MMP-9与TIMP-1及TGF-β1,光镜下观察肺脏病理组织形态。
结果：肺组织病理切片显示,活血定喘汤能显著减少炎性细胞的浸润,肺泡壁水肿及支气管平滑肌的增厚,能抑制哮喘气道重塑的发生；血浆检测显示模型组及治疗各组与正常组相比MMP-9与TIMP-1的水平均明显升高,经治疗后均有降低,其中西药组和活血定喘汤高剂量组与模型组之间有明显差异(P<0.01)；同时除正常组之外,其余各组都存在着MMP-9/TIMP-1比例的失衡,模型组MMP-9/TIMP-1的比值出现倒置失衡；血浆TGF-β1的检测结果显示：与正常组比较模型组中TGF-β1的含量显著升高,两组之间存在明显差异(P<0.01),经治疗后高剂量组及西药组上述指标均显著降低,与模型组之间有明显差异(P<0.01),而高剂量组及西药组作用之间无明显差异。
结论：活血定喘汤能抑制哮喘气道重塑的进程,其部分机制可能通过调节哮喘气道重塑小鼠血浆MMP-9、TIMP-1、TGF-β1的水平及MMP-9/TIMP-1之间的平衡实现的。
Objective:Observed the relative content of plasma MMP-9 and TIMP-1, the change of plasma TGF-β1 and lung morphology in Balb/c mice, so as to explore the effect and mechanism of Huoxue Dingchuan Soup(HDS) to MMP-9/TIMP-1 balance and asthmatic airway remodeling.
Methods:Balb/c mice 70 were randomly divided into normal group, model group, western medicine group, HDS low-dose group and high-dose group. Asthmatic airway remodeling model was established by repeated inhalation of ovalbumin. The normal group and model group administrated water, the other groups respectively administrated the corresponding medicines. At the end, determined plasma MMP-9,TIMP-1 and TGFβ1 by ELISA, observed the pathomorphology of lung under light microscope.
Results::The pathological changes in lung tissue showed that HDS can significantly reduce the infiltration of inflammatory cells, alveolar wall edema and thickening of bronchial smooth muscle, can inhibit the occurrence of airway remodeling; model group compared with the control group, the level of MMP-9 and TIMP-1 except the normal group were significantly increased, after treatment they were significantly decreased, in which western medicine group and HDS high-dose group have significant difference comparing with the model group (P<0.01); In addition to the normal group at the same time, the other groups of MMP-9/TIMP-1 are imbalance in the ratio, the model group of TIMP-1 over-proportional increase eventually lead to an imbalance; plasma TGF-β1 results showed that compared with the normal group, TGF-β1 levels of model group were significantly increased, there is significant difference between the two groups (P<0.01), after treatment, the above indices of high-dose group and western medicine group decreased significantly and were significant differences compared with the model (P <0.01), while the high-dose group and western medicine group were no significant difference.
Conclusion:HDS might suppress asthmatic airway remodeling by regulating the level of MMP-9, TIMP-1, TGF-β1 in the lung tissue of asthmatic airway remodeling mice and MMP-9/TIMP-1 balance.
引文
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