归芪白术方含药血清对胃癌MKN-45细胞增殖和IL-17、NF-κB表达的影响  

Effects of Guiqibaizhufang serum on proliferation and IL-17,NF-κB expression of gastric cancer MKN-45 cells

作　　者：苏韫 李春波 龚红霞 薛轩 李婷婷 牛世伟 SU Yun;LI Chun-Bo;GONG Hong-Xia;XUE Xuan;LI Ting-Ting;NIU Shi-Wei(School of Basic Medicine,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学基础医学院,兰州730000 [2]甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000

出　　处：《中国免疫学杂志》2021年第15期1824-1829,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(81660744);甘肃省高等学校创新基金项目(2020A-072);甘肃省自然科学基金项目(1508RJZA047)。

摘　　要：目的:探讨归芪白术方含药血清通过IL-17/NF-κB信号通路对胃癌MKN-45细胞增殖的影响。方法:制备生药质量浓度6.12 g/ml的归芪白术方药液,将SD大鼠随机分为空白组(蒸馏水),阳性药组(奥沙利铂),归芪白术方高、中、低剂量组(6.12 g/ml、3.06 g/ml、1.53 g/ml),灌胃后制备归芪白术方含药血清,用10%含药血清干预MKN-45细胞,通过细胞增殖及细胞毒性实验(CCK-8)检测归芪白术方含药血清对MKN-45细胞增殖的影响;通过Western blot检测IL-17、NF-κBp65、p-NF-κBp65、IKK-β和IL-6蛋白的表达情况;通过实时荧光定量聚合酶链式反应(Real-time PCR)检测IL-17、NF-κBp65、IKK-β和IL-6 mRNA表达的情况。结果:CCK-8实验显示,与空白组比较,归芪白术方含药血清抑制MKN-45细胞增殖,且呈时间与剂量依赖性(P<0.05)。Western blot结果显示,与空白组比较,阳性药组和归芪白术方高、中、低剂量含药血清组IL-17、NF-κBp65、p-NF-κBp65、IKK-β和IL-6蛋白表达下降(P<0.05);Real-time PCR实验表明,与空白组比较,阳性药组和归芪白术方高、中、低剂量含药血清组IL-17、NF-κBp65、IKK-β和IL-6 mRNA表达减少(P<0.05)。结论:归芪白术方含药血清可以抑制胃癌MKN-45细胞的增殖,可能与IL-17/NF-κB信号通路有关。Objective:To explore the effect of serum containing Guiqibaizhufang on the proliferation of gastric cancer MKN-45 cells through IL-17/NF-κB signaling pathway.Methods:To prepare crude drug solution with a mass concentration of 6.12 g/ml,and SD rats were randomly divided into blank group(distilled water),positive drug group(oxaliplatin),and Guiqibaizhufang high-dose,medium-dose and low-dose groups(6.12 g/ml,3.06 g/ml,1.53 g/ml).After intragastric administration,drug-containing serum of Guiqibaizhufang was prepared.10%drug-containing serum was used to intervene MKN-45 cells.Cell proliferation and cytotoxicity test(CCK-8)was used to detect the effect of drug-containing serum of Guiqibaizhufang on MKN-45 cells proliferation.The protein expressions of IL-17,NF-κBp65,p-NF-κBp65,IKK-βand IL-6 were detected by Western blot.Real-time PCR was used to detect the expression of IL-17,NF-κBp65,IKK-βand IL-6 mRNA.Results:The CCK-8 experiment showed that compared with the blank group,the serum containing Guiqibaizhufang inhibited the proliferation of MKN-45 cells in a time and dose dependent manner(P<0.05).Western blot results showed that compared with blank group,the expressions of IL-17,NF-κBp65,pNF-κBp65,IKK-βand IL-6 were decreased in positive drug group and Guiqibaizhufang high-dose,medium-dose and low-dose drug-containing serum groups(P<0.05).Real-time PCR assay showed that compared with blank group,the mRNA expressions of IL-17,NF-κBp65,IKK-βand IL-6 were decreased in positive drug group and Guiqibaizhufang high-dose,medium-dose and low-dose drug-containing serum groups(P<0.05).Conclusion:The serum containing Guiqibaizhufang can inhibit the proliferation of MKN-45 cells in gastric cancer,which may be related to the IL-17/NF-κB signaling pathway.

关 键 词：归芪白术方 含药血清 胃癌MKN-45细胞 IL-17/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]