肺岩宁颗粒对Lewis肺癌小鼠肿瘤生长及肿瘤相关巨噬细胞的影响

Affect of Feiyanning granule on Lewis lung carcinoma growth and tumor associated macrophages

目的观察肺岩宁颗粒对Lewis肺癌小鼠肿瘤生长的抑制作用,并探索其对肿瘤相关巨噬细胞(TAM)的影响。方法由C57BL/6小鼠造模成Lewis肺癌移植瘤模型,随机分为3组:模型组、顺铂组和肺岩宁颗粒组,每组各8只。药物干预14 d后观察各组小鼠生存质量和抑瘤率;实时荧光定量聚合酶链反应(RT-q PCR)检测肿瘤组织中基质金属蛋白酶(MMP)-2、MMP-9、信号转导和转录激活因子(STAT)3以及核转录因子(NF)-κB p65 m RNA的表达;流式细胞术检测肿瘤组织中M_1及M_2比例;蛋白质印迹法检测肿瘤组织中磷酸化STAT(p-STAT)3的表达。结果三组间比较,肺岩宁颗粒组小鼠生存质量较好,同时也显示了抑制肿瘤生长的作用。流式细胞术检测发现,与模型组比较,顺铂组、肺岩宁颗粒组M_2的表达降低(P<0.05);与模型组及顺铂组比较,肺岩宁颗粒组M_1增加,且差异有统计学意义(P<0.05)。RT-q PCR检测发现,与模型组比较,顺铂组、肺岩宁颗粒组肿瘤组织中MMP-2、MMP-9和NF-κB p65 m RNA表达均降低,且差异均具有统计学意义(P<0.05);肺岩宁颗粒组与顺铂组两者比较,前者表达更低(P<0.05)。蛋白质印迹法检测发现,与模型组比较,顺铂组、肺岩宁颗粒组细胞中p-STAT 3表达均有降低趋势。结论肺岩宁颗粒具有抑制肺癌肿瘤细胞生长的作用,该作用可能与经NF-κB、STAT 3信号通路干预TAM的作用有关。

Objective To observe the effects of Feiyanning granule on the tumor associated macrophages（TAMs） and suppressing growth of Lewis lung carcinoma in mice. Methods The tumor-bearing model was established by subcutaneously injecting Lewis cells to C57 BL/6 mice. Then the C57 BL/6 mice were divided into model group（n=8）, cisplatin group（n=8）, and Feiyanning granule group（n=8）, and treated with corresponding drugs. The quality of life and the tumor inhibition rate were observed after 14 days. The m RNA expression of MMP-2, MMP-9, NF-κB p65 and STAT 3 were detected by realtime quantitative PCR（RT-q PCR）. The ratios of M1 and M2 were measured by flow cytometry method. The expression of p-STAT 3 protein was detected by Western-Blot. Results According to the quality of life, the Feiyanning granule group was better. Feiyanning granule was able to suppress Lewis lung carcinoma growth. Compared with model group, the M2 of Feiyanning granule group and cisplatin group were significantly lower（P〈0.05）. Compared with model group and cisplatin group, the M1 of Feiyanning group was significantly higher（P〈0.05）. The result of RT-q PCR showed that the expressions of MMP-2, MMP-9 and NF-κB p65 m RNA in both cisplatin group and Feiyanning granule group were obviously lower than those in model groups（P〈0.05）, and the expressionsin Feiyanning granule group were lower than those in cisplatin group（P〈0.05）. The result of Western-Blot showed that the protein expressions of p-STAT 3 in both Feiyanning granule group and cisplatin group was lower than those in model group. Conclusion Feiyanning granule can suppress Lewis lung carcinoma growth. Its mechanisms may be related to the intervention of TAMs through NF-κB and STAT 3 signal pathway.