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

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.