复方中药对肺纤维化小鼠肺组织COL-Ⅰ、Caspase-11及细胞因子水平的影响

Influence of Compound TCM on COL-Ⅰ and Caspase-11 and cytokine in Lung Tissues and of Pulmonary Fibrosis Mice

目的:观查中药复方对肺纤维化小鼠肺组织COL-Ⅰ、Caspase-11及细胞因子IL-1β、TNF-α、LPS表达水平的影响,探讨复方中药对肺纤维化的治疗作用及可能的作用机制。方法:清洁级昆明小鼠48只随机分为六组(每组8只):空白对照组、模型组、中药复方一组(丹参组)、中药复方二组(复方组)。除空白对照组外,其余各组小鼠给予2.5 mg/kg LPS生理盐水溶液(1 mg/m L)滴鼻诱导肺纤维模型,造模后3 d,药物组每天分别给予相应药物剂量灌胃,21天后,全部处死,取小鼠肺部组织,ELISA方法检测IL-1β、TNF-α、LPS表达含量,Western blot检测Ⅰ型胶原蛋白COL-Ⅰ、蛋白酶Caspase-Ⅱ含量表达。结果:Western blot检测结果:与空白组相比,各组COL-Ⅰ的灰度值均升高,但差异无统计学意义(P>0.05),与模型组相比,各药物组COL-Ⅰ的灰度值均有下降趋势,其中醇提组较显著(P>0.05);Caspase-11的表达,模型组与各药物组间均高于空白组,其中模型组与丹参组升高比较明显(P<0.05),与模型组相比较,各药物组Caspase-11的表达均有下降趋势,其中复方组的表达较显著,差异有统计学意义(P<0.05)。细胞因子检测结果:与空白组相比,各组小鼠肺组织IL-1β、TNF-α、LPS水平明显升高(P<0.05);与模型对照组相比,各药物组IL-1β、TNF-α、LPS含量均明显下降(P<0.05)。结论:各组复方药物均能有效降低肺纤维化组织IL-1β、TNF-α、LPS的表达水平,Caspase-11作为最新研究发现参与炎症反应、细胞凋亡的蛋白,本次实验中证明,Caspase-11参与肺纤维化的形成,且中药复方组与醇提组能够有效降低其表达量,其可能为减轻肺纤维化形成的机制。

Objective： To investigate the therapeutic effect of compound traditional Chinese medicine on pulmonary fibrosis and its probable mechanism through examining the effect of compound traditional Chinese medicine on expression levels of lung tissues of mice with pulmonary fibrosis： tumor necrosis factor（TNF-α）,interleukin-1β（IL-1β） and LPS. Methods： Forty-eight Kunming mice of clean grade were randomly divided into six groups（8 of each group） ： the control group,the untreated group and the Chinese herbs treatment group,which were subdivided into four different groups,the Danshen group,the compound group,the water extraction group and the alcohol extraction group. The untreated group and the Chinese herbs treatment group respectively received the nasal cavity injection 2. 5 mg/kg LPS normal saline（1 mg/mL） to make the pulmonary fibrosis mice models. Three days after the formation of the models,the Chinese herbs treatment groups were given corresponding drugs respectively by using stomach tubes. 21 days later,we killed all mice to take the lung tissues of them. And then we examined the lung tissue pathology levels of TNF-α,IL-1β and LPS of each group by means of ELISA method and the levels of Type I collagen（COL-I）,protease and Caspase-11 by using Western blot method. Results： Compared with the untreated group,levels of TNF-α,IL-1β and LPS of each treatment group were decreased strikingly（P〈 0. 05）. Test results of Western blot method： compared with the control group,the expressions of Gray value in COL-I of the untreated group and all treatment groups were all increased（P〉0. 05）.Compared with the untreated group,the expressions of Gray value in COL-I of all treatment groups all had a tendency of decrease,among which the alcohol extraction group was more obvious（P〉0. 05）. As for the level of Caspase-11,the untreated group and all treatment groups＇ were higher than the control group,and particularly,the untreated group and the Danshen group＇s was increased more obviously（P〈 0. 05）. Compared with the untreated group,the levels of Caspase-11 of each treatment group all tended to decrease,the Danshen group and the alcohol extraction group in particular（P〈 0. 05）. Conclusion： All Chinese herbs in each treatment group can effectively reduce the secretion of the relevant cytokines leading to pulmonary fibrosis and mitigate the formation of pulmonary fibrosis. Its mechanism is probably related to the activation of the apoptotic protein Caspase-11 which is hindered by drug treatment.