以NLRP3炎症小体为靶点的补肺消积饮治疗气阴两虚型肺癌作用机制研究

NLRP3 inflammasome played an important role in tumour progression and had became a new research hotspot. However, the study to treat lung carcinoma targeted by NLRP3 inflammasome is scarce at present. The proinflammatory factors, such as Interleukin-lβ(IL-1β), et al，were important triggers of lung cancer with deficiency of Qi and Yin as well as the major productions of NLRP3 inflammasome. Therefore, we supposed that there was a substantial connection between NLRP3 inflammasome and lung cancer with deficiency of Qi and Yin, which was not only NLRP3 inflammasome can promote the progress of lung cancer with deficiency of Qi and Yin，but also the deficiency of Qi and Yin in lung cancer can accelerate the expression of NLRP3 inflammasome. To verify this hypothesis, non-small cell lung cancer cell line A549 was selected in this study. CCK-8 assay was used to evaluated the viability of cells. ELISA、qRT-PCR、Western Blot analysis were applied to determine the expression of NLRP3 inflammasome and its productions. Immunohistological staining was performed to observe the pathology of tomor tissues in the nude mouse model of lung cancer with deficiency of Qi and Yin. Meanwhile, in order to provide theoretical and experimental basis in the prevention and treatment of lung cancer targeted by NLRP3 inflammasome used traditional chinese medicine, bufeixiaojiyin recipe which had the effect of benefiting qi and nourishing Yin was shose to illuminate one of the mechanisms to treat lung cancer with deficiency of Qi and Yin was inhibiting NLRP3 inflammasome.

NLRP3炎症小体对肿瘤具有重要的促进作用，已成为研究的新热点。然而目前以其为靶点的肺癌治疗研究还比较少。IL-1β等促炎因子是肺癌-气阴两虚证的重要指征，也是NLRP3炎症小体的主要产物，因此，我们提出NLRP3炎症小体与肺癌-气阴两虚证密切相关, NLRP3炎症小体能够促进肺癌-气阴两虚证的进展，而肺癌中“气阴两虚状态”亦可以促进NLRP3炎症小体表达的假说。本研究选取非小细胞系肺癌细胞A549作为株种，用CCK-8测定细胞存活率，ELISA、qRT-PCR、Western Blot检测NLRP3炎症小体及其产物的表达，免疫组织化学法染色观察裸鼠肺癌-气阴两虚证模型肿瘤组织病理表现来验证这个假说，同时阐明具有益气养阴功效的验方补肺消积饮治疗气阴两虚型肺癌的机制之一是通过阻断NLRP3炎症小体发挥作用的,为以NLRP3为靶点的中医药防治肺癌提供理论和实验依据。

NLRP3炎症小体对肿瘤具有重要的促进作用，其不仅是炎症发生的中心环节，还能够促进肿瘤形成、侵袭、转移，目前已成为肿瘤治疗研究的新热点。然而目前以其为靶点的肺癌治疗研究还比较少。IL-1β等促炎因子是肺癌-气阴两虚证的重要指征，也是NLRP3炎症小体的主要产物，因此，我们提出NLRP3炎症小体与肺癌-气阴两虚证密切相关，NLRP3炎症小体能够促进肺癌-气阴两虚证的进展，而肺癌中“气阴两虚状态”亦可以促进NLRP3炎症小体表达的假说。本研究选取非小细胞系肺癌细胞A549作为株种，用CCK-8测定细胞增殖率，流式细胞术测定细胞凋亡率，ELISA、qRT-PCR、Western Blot检测NLRP3炎症小体及其产物的表达，免疫组织化学法染色观察裸鼠肺癌-气阴两虚证模型肿瘤组织NLRP3炎症小体及其组分的组织学特征。本研究的实验结果显示：1.体外实验结果提示：不同浓度的补肺消积饮含药血清能够通过对人肺腺癌A549细胞进行增殖抑制和诱导凋亡作用，尤其以高浓度作用最为显著，从而降低细胞中炎症因子IL-1β及1L-18的表达水平，改善肺癌A549细胞的炎症微环境，以起到治疗的作用。2.体内实验结果提示：补肺消积颗粒能有效改善肺癌气阴两虚证裸鼠的一般生活状态和肿瘤生长情况，降低气阴两虚评价因子MDA、IL-1β、IL-18以及NLRP3炎症小体的表达。本研究结果验证了NLRP3炎症小体能够促进肺癌-气阴两虚证的进展，而肺癌中“气阴两虚状态”亦可以促进NLRP3炎症小体表达的假说，同时也阐明具有益气养阴功效的验方补肺消积饮治疗气阴两虚型肺癌的机制之一是通过阻断NLRP3炎症小体发挥作用的，一定程度上为以NLRP3为靶点的中医药防治肺癌提供了理论和实验依据。