To investigate the effect of jianpi-jiedu (JPJD) prescription-contained serum on colorectal cancer SW48 cell proliferation and the underlying mechanisms. Methods: Crude extract from JPJD was made by water extract method and the main components of crude extract from JPJD were analyzed by ultra-performance liquid phase high resolution time of flight mass spectrometry (UPLC-Q-TOF/MS). The low, medium, and high-concentration of JPJD-contained serum were prepared by the serum pharmacological method. The effect of serum containing JPJD on SW48 cell proliferation was determined by MTT assay. The cell cycle was detected by flow cytometric method. The protein levels of mammalian target of rapamycin (mTOR), phospho-mTOR, P-P53, and -P21, and the mRNA level of mTOR were examined by Western blot and RT-PCR, respectively. Results: Seven compounds including calycosin-7-glucoside, astragaloside, ginsenoside-Re, ginsenoside-Rb1, glycyrrhizinic acid, apigenin, atractylenolide-II were identified. MTT assays demonstrated that the SW48 cell proliferation was inhibited by medium and high concentration of JPJD-contained serum and the percentages of cells at G1 phase in SW48 cell cultured in the medium and high concentration of JPJD serum group were significantly higher than those in the control group (P<0.05). Meanwhile, the levels of mTOR mRNA and phospho-mTOR protein in the medium and high concentration of JPJD serum groups were substantially lower than those in the control group (P<0.05). Conversely, the expressions of phospho-P53 and P21 protein were significantly increased in the medium and high concentration of JPJD serum group compared with those in the control group. Conclusion: JPJD prescription-contained serum can inhibit SW48 cell proliferation, which may be related to mTOR-P53-P21 signaling pathways.
目的：研究健脾解毒方含药血清对人大肠癌SW48细胞增殖的影响及作用机制。方法：采用水提法制作健脾解毒方粗提取物，超高效液相-高分辨飞行时间质谱法分析健脾解毒方粗提取物的主要活性成分，利用血清药理学方法制备低、中、高浓度健脾解毒方含药血清，MTT法检测人结肠癌SW48细胞增殖活性，运用流式细胞术检测细胞周期，RT-PCR技术检测哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin，mTOR) mRNA表达，Western 印迹技术检测磷酸化(phospho，P)-mTOR，P-P53和P21的蛋白表达。结果：通过高分辨质谱结合对照品数据相关分析，对健脾解毒方粗提取物中7个主要成分(毛蕊异黄酮苷、黄芪甲苷、人参皂苷Re、人参皂苷Rb1、甘草酸、芹菜素、白术内酯II)进行了鉴定及分析；健脾解毒方中、高剂量含药血清对SW48细胞有明显抑制增殖作用；健脾解毒方中、高剂量含药血清使SW48细胞周期阻滞在G1期；健脾解毒方含药血清能够下调的mTOR mRNA及P-mTOR蛋白表达，上调P-P53和P21蛋白的表达。结论：健脾解毒方可能通过mTOR-P53-P21途径抑制人结肠癌SW48细胞的增殖，这可能是健脾解毒方治疗结肠癌的作用机制之一。.