背景:中医(TCM)是在其独特的理论体系的指导下临床使用的。补肺益神(BY)颗粒,口服中草药配方,证实对治疗慢性阻塞性肺疾病(COPD)患者的肺肾气虚证有效。舒肺贴软膏是另一种穴位贴敷(AS)疗法的处方,其原理是通过适当的穴位外治来治疗内部疾病。BY颗粒联合舒肺贴的有益效果已在先前的临床试验中得到证明。然而,潜在机制尚不清楚.本研究旨在探索BY颗粒和穴位贴敷综合疗法通过调节香烟烟雾/细菌暴露诱导的COPD大鼠模型中过氧化物酶体增殖物激活受体-γ(PPARγ)信号传导的抗氧化机制。
方法:大鼠随机分为对照组,型号,BY,AS,BY+AS和氨茶碱(APL)组。COPD大鼠是由香烟烟雾和细菌暴露引起的,并服用生理盐水,BY颗粒,AS,BY+AS或氨茶碱从第9周开始并在第20周处死。通过羟胺和硫代巴比妥酸法测定外周血和支气管肺泡灌洗液(BALF)中超氧化物歧化酶(SOD)的活性和甲烷二羧酸(MDA)的水平。采用定量聚合酶链反应和免疫印迹法检测肺组织中PPARγ基因和蛋白的表达。
结果:模型组血清和BALFSOD明显降低(P<0.01),MDA升高(P<0.01)。与COPD大鼠相比,各治疗组血清SOD均较高(P<0.01),BY和BYAS组BALFSOD较高(P<0.01);所有治疗组血清和BALFMDA较低(P<0.01)。BY+AS组血清和BALFSOD高于AS组,MDA含量较低(P<0.05)。BALF+AS组较APL组SOD升高,MDA降低(P<0.05)。COPD大鼠PPARγmRNA和蛋白及PPARγ磷酸化(p-PPARγ)水平降低(P<0.01),各治疗组升高(P<0.01)。BY+AS组PPARγmRNA高于AS组(P<0.05),BY+AS组PPARγ和p-PPARγ高于AS组和APL组(P<0.05,P<0.01);BY组PPARγ蛋白高于APL组(P<0.05)。
结论:补肺益肾颗粒,舒菲铁及其组合对稳定期COPD有有益作用,可以减弱氧化应激,PPARγ信号的激活可能参与了潜在的机制,但补肺益肾颗粒与舒肺贴无明显协同作用。
BACKGROUND: Traditional Chinese medicine (TCM) is clinically used under the guidance of its unique theory system. Bufei Yishen (BY) granules, an oral Chinese herbal formula, is confirmed effective for treating the syndrome of lung-kidney qi deficiency in chronic obstructive pulmonary disease (COPD) patients. Shu-Fei Tie ointment is another prescription for acupoint sticking (AS) therapy based on the theory of treating an internal disease by external treatment on proper acupoints. The beneficial effects of BY granules combined with Shu-Fei Tie have been proved in previous clinical trials. However, the underlying mechanism remains unclear. The present study was initiated to explore the antioxidative mechanism of the integrated therapy of BY granules and acupoint sticking via regulating by peroxisome proliferator activated receptor-gamma (PPARγ) signaling in a cigarette-smoke/bacterial exposure induced COPD rat model.
METHODS: Rats were randomized into Control, Model, BY, AS, BY+AS and aminophylline (APL) groups. COPD rats were induced by cigarette-smoke and bacterial exposures, and were administrated with normal saline, BY granules, AS, BY+AS or aminophylline from week 9 and sacrificed at week 20. Activity of superoxide dismutase (SOD) and levels of methane dicarboxylic aldehyde (MDA) in peripheral blood and bronchoalveolar lavage fluid (BALF) were determined by hydroxylamine and thiobarbituric acid methods. The gene and protein expressions of PPARγ in the lung tissues were analyzed by quantitative polymerase chain reaction and western blot.
RESULTS: Serum and BALF SOD decreased significantly in Model group (P<0.01), while MDA increased (P<0.01). Compared to COPD rats, serum SOD was higher in all treatment groups (P<0.01), and BALF SOD was higher in BY and BY+AS groups (P<0.01); serum and BALF MDA was lower in all treatment groups (P<0.01). Serum and BALF SOD was higher in BY+AS group than in AS group, while MDA was lower (P<0.05). BALF SOD increased in BY+AS group compared with APL group, while MDA decreased (P<0.05). PPARγ mRNA and protein and the phosphorylation of PPARγ (p-PPARγ) decreased in COPD rats (P<0.01), and increased in all treatment groups (P<0.01). PPARγ mRNA was higher in BY+AS group than in AS group (P<0.05), PPARγ and p-PPARγ were higher in BY+AS group than in AS and APL groups (P<0.05, P<0.01); PPARγ protein was higher in BY group than in APL group (P<0.05).
CONCLUSIONS: Bufei Yishen granules, Shu-Fei Tie and their combination have beneficial effects in stable COPD, and can attenuate the oxidative stress, and the activation of PPARγ signaling might be involved in the underlying mechanisms, but there are no obvious synergistic effect of Bufei Yishen granules and Shu-Fei Tie.