加味茵陈蒿汤对湿热哮喘大鼠气道白细胞介素-6,信号转导因子-3表达的影响及与气道炎症的关系

Influence of Jiawei Yinchenhao Decoction on Expression of IL-6/STAT3 and Relationship Between IL-6,STAT3 and Acute Airway Inflammation in Dampness-heat Asthma Rat

目的:探讨加味茵陈蒿汤对湿热哮喘大鼠气道急性炎症的影响及其可能的机制。方法:50只健康雄性SD大鼠随机分成5组,每组10只,分别为空白组、湿热哮喘模型组、加味茵陈蒿汤低、高剂量组(6.29,12.58 g·kg-1·d-1)、地塞米松组(0.08 mg·kg-1·d-1)。多因素干预建立湿热哮喘大鼠模型,实验第16天开始给药,末次激发24 h以内处死大鼠,腹主动脉采血,留取肺组织检测。对各组大鼠支气管肺泡灌洗液(BALF)中细胞进行计数和炎性细胞分类,酶联免疫吸附实验(ELISA)法测定BALF中细胞因子白细胞介素(IL)-2,IL-4,IL-5,IL-6,干扰素-γ(IFN-γ)水平,免疫组织化学染色检测气道信号转导因子-3(STAT3)蛋白表达,以甘油醛-3-磷酸脱氢酶(GAPDH)为内参,RT-PCR定量检测肺组织STAT3水平。应用SPSS 17.0软件进行统计分析。结果:与正常组比较,模型组IL-4,IL-5,IL-6水平升高(P<0.05),IL-2,IFN-γ水平降低(P<0.05),气道STAT3蛋白、肺组织STAT3 mRNA水平升高;与模型组比较,各给药组大鼠IL-4,IL-5,I L-6水平降低(P<0.05),IL-2,IFN-γ水平均升高(P<0.05),且哮喘大鼠BALF中IL-6,气道STAT3蛋白,肺组织STAT3 mRNA分别与BALF中细胞总数及嗜酸性粒细胞,淋巴细胞,IL-4,IL-5,IL-6呈正相关(P<0.01),与IL-2,IFN-γ呈负相关(P<0.01)。各给药组均可降低气道炎症及STAT3信号通路表达,地塞米松组疗效最优,其次为加味茵陈蒿汤低剂量组。结论:加味茵陈蒿汤能明显降低气道炎症,其作用机制可能与抑制IL-6/STAT3信号通路表达从而调节Thl/Th2细胞因子失衡有关。

Objective： To discuss the effect of Jiawei Yinchenhao decoction on the acute airway inflammation and its possible mechanism in the of dampness-heat asthma rat. Method： Fifty healthy male SD rats were randomly divided into five groups： control group,dampness-heat asthma model group and Jiawei Yinchenhao decoction low dose group,Jiawei Yinchenhao decoction high dose group,dexamethasone group,10 rats in each group. The dampness-heat asthma model was induced by multi-factors. The rats in dampness-heat asthma groups were fed with medicine from 16 th day of the experiment. The rats were killed within 24 h after the last asthma provocation test to collect abdominal aortic blood and lung tissue specimens for testing indexes. The bronchoalveolar lavage fluid（ BALF） was collected,the total cell and the cell differentials were counted,the level of interleukin-2（ IL-2）,IL-4,IL-5,IL-6,and interferon-γ（ IFN-γ） in BALF were measured by ELISA. The expression of signal transducers and activators of transcription 3（ STAT3） in airway was detected by immunohistochemistry technique. With glyceraldehyde-3-phosphate dehydrogenase（ GAPDH） being a control,airway tissue extracts were used for detecting STAT3 / GAPDH expression level by reverse transcription-polymerase chain reaction（ RTPCR）. SPSS 17. 0 software was used to analyze the data. Result： Compared with the control group,the level of IL-2 and IFN-γ in BALF of the other groups was declined while IL-4,IL-5 and IL-6 level was higher than that in the control group. Compared with the dampness-heat asthma group,the IL-4,IL-5 and IL-6 level in treatment groups was significantly lower than that in the model group（ P 〈 0. 05）,while IL-2,IFN-γ was higher. The level of IL-6,STAT3 and STAT3 / GAPDH had positive correlations with IL-4,IL-5 and IL-6 in BALF（ P 〈 0. 05） and negative correlations with IL-2,IFN-γ. All the treatment group could reduce airway inflammation. Conclusion：Clearing heat and removing dampness and phlegm mothed could reduce the acute airway inflammation,its efficacy is dose-related,the Chinese medicine low dose group has better effect. Its mechanism may be related to inhibition of IL-6 / STAT3 signaling pathway to regulate the expression of Thl / Th2 cytokine imbalance.