西黄丸对炎症微环境下肺癌A549荷瘤小鼠NLRP3炎症小体及其产物和肿瘤增殖的影响

Effect of Xihuangwan on NLRP3 Inflammatory Bodies and Their Products and Tumor Proliferation of Lung Cancer A549 Bearing Nude Mice in Inflammatory Microenvironment

目的:研究并探讨西黄丸对炎症微环境下的肺癌A549荷瘤小鼠的抑瘤作用,并探讨西黄丸对肺癌A549荷瘤小鼠的血清及肿瘤组织中的核苷酸寡聚化结构域样受体蛋白3(NLRP3)炎症小体及其产物表达的影响作用。方法:采用肺癌A549荷瘤小鼠模型,肺癌A549细胞采用培养基中加入脂多糖(LPS)+三磷酸腺苷(ATP)的办法造炎症微环境下的肺癌A549细胞模型。造模成功后随机分为空白组(等体积生理盐水),MCC950组(MCC950溶液,0.79 g·kg^-1),西黄丸低、中、高剂量组(0.39,0.78,1.95 g·kg^-1),采用口服给药的方式进行,每天给药1次,连续给药21 d,处死小鼠并剥离肿瘤组织测量肿瘤体积及质量,免疫组织化学法检测肿瘤组织NLRP3表达,蛋白免疫印迹法(Western blot)检测丙二醛(MDA),白细胞介素-1β(IL-1β),白细胞介素-18(IL-18),NLRP3炎性小体蛋白表达,眼底静脉取血以酶联免疫吸附试验(ELISA)检测血清IL-1β,IL-18,MDA表达水平。结果:与空白组比较,MCC950组,西黄丸低、中、高剂量组的抑瘤率分别为39.21%,31.72%,42.24%,55.68%;ELISA检测发现,西黄丸高剂量组能明显下调小鼠血清中的MDA,IL-1β,IL-18表达水平(P<0.05);Western blot结果显示,与空白组比较,西黄丸高剂量组能够有效降低小鼠肿瘤组织中的MDA,IL-1β,IL-18,NLRP3的蛋白表达水平(P<0.05);免疫组织化学法结果显示,与空白组比较,MCC950组与西黄丸各剂量组均能降低小鼠肿瘤组织中的NLRP3炎症小体表达阳性率(P<0.05),且MCC950组和高剂量西黄丸组抑制效果最好(P<0.05)。结论:西黄丸可以抑制荷肺癌A549细胞小鼠的肿瘤组织生长,其机制可能是通过抑NLRP3炎症小体及IL-1β,IL-18,MDA的表达水平,进而抑制肿瘤细胞的炎症微环境达到抑制肿瘤细胞生长的结果。

Objective:To study the antitumor effect of Xihuangwan on A549 lung cancer nude mice in inflammatory microenvironment,and explore the effect of Xihuangwan on the expressions of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammatory bodies and their products in serum and tumor tissue of A549 lung cancer nude mice.Method:The lung cancer A549 cell model was established in nude mice with lung cancer,and the lung cancer A549 cell model was established in inflammatory microenvironment by adding lipopolysaccharide(LPS)+adenosine triphosphate(ATP)to the culture medium.After modeling,the rats were randomly divided into blank group(equal volume of normal saline),positive drug control group(MCC950 solution,0.79 g·kg^-1),and low,medium,high-dose Xihuangwan groups(0.39,0.78,1.95 g·kg^-1).The rats were administered orally once a day for 21 days,and then sacrificed.The tumor tissues were stripped to measure the tumor body.The expressions of NLRP3,malondialdehyde(MDA),interleukin(IL)-1β,IL-18 and NLRP3 were detected by Western blot,and the levels of IL-1β,IL-18 and MDA were detected by enzyme linked immunosorbent assay(ELISA).Result:Compared with the blank group,the tumor inhibition rates in the positive drug control group and the low,medium and high dose Xihuangwan group were 39.21%,31.72%,42.24%and 55.68%.ELISA showed that the high-dose Xihuangwan group could significantly reduce the expressions of MDA,IL-1βand IL-18 in the serum of nude mice(P<0.05).Western blot showed that the highdose Xihuangwan group could effectively reduce the protein expressions of MDA,IL-1β,IL-18 and NLRP3 in tumor of nude mice(P<0.05),the results of immunohistochemistry showed that the expression rate of NLRP3 in the tumor tissues of nude mice was reduced in the positive drug group and each dose of Xihuangwan group(P<0.05).Conclusion:Xihuangwan can inhibit the growth of tumor tissue of A549 cells bearing lung cancer in nude mice.The mechanism may be that it can inhibit the growth of tumor cells by inhibiting the expression of NLRP3 inflammatory bodies,IL-1β,IL-18,MDA tables,and then inhibiting the inflammatory microenvironment of tumor cells.