摘要
目的探讨李忠教授基于厥阴理论辨证论治恶性肿瘤的核心处方,并进行中药含药血清对肿瘤细胞抑制作用的体外实验验证。方法收集李忠教授北京中医药大学东直门医院国际部门诊2022年1月至2023年6月恶性肿瘤患者的处方数据,通过古今医案云平台V 2.3.7分析药物频次、性味归经,并进行中药间的聚类分析、关联分析、复杂网络分析,总结核心处方。将8只SPF级6~8周龄SD雄性大鼠按照随机数字表法将其分为实验组、对照组,各4只。分别以2 ml核心处方中药煎剂与同剂量生理盐水灌胃中药组和对照组,连续灌胃7 d,提取含药血清。CCK8法确定含药血清对A549、HepG2细胞的最佳干预浓度与干预时间。设置空白组:完全培养基培养,不加药;中药组:以最佳浓度含药血清进行干预;对照组:以同剂量空白血清进行干预。Transwell实验检测细胞迁移及侵袭能力,流式细胞术进行细胞凋亡及周期检测,Western blot检测细胞凋亡与上皮细胞-间充质转化相关蛋白的表达。结果共纳入8166首处方,药物共300种,用药性温、味甘较多,归经以肝、肺经为多。黄芪、鳖甲、乌梅、山萸肉等16味药物使用频率均>90%。结合聚类分析、复杂网络分析等结果与李忠教授经验,得出平调阴阳法核心处方1首,包括生黄芪、炙黄芪、醋鳖甲、仙鹤草、乌梅、党参、当归、熟地黄、忍冬藤、白芍、穿山龙、浙贝母、僵蚕、生牡蛎、山萸肉。实验研究发现,中药含药血清最佳干预浓度为20%、干预时间为48 h。中药组A549、HepG2细胞的迁移和侵袭能力低于对照组,细胞凋亡率高于对照组,阻滞细胞周期长于对照组,Bcl-2、N-Cadherin、Vimentin表达水平低于对照组,Cleaved-Caspase-3、E-Cadherin表达水平高于对照组(P<0.05)。结论运用数据挖掘可得出从厥阴论治恶性肿瘤的核心处方,处方含药血清可体外抑制肿瘤细胞的增殖、迁移与侵袭,机制可能与促进细胞凋亡、阻滞细胞周期、抑制上皮细胞间充质转化相关。
Objective To explore the core prescription of Professor Li Zhong’s treatment of malignant tumors based on Jueyin theory and carry out in experimental verification of the inhibitory effect of drug-containing serum on tumor cells.Methods Professor Li Zhong’s prescription data of patients with tumors from January 2022 to June 2023 in the Department of International,Dongzhimen Hospital,Beijing University of Chinese Medicine were collected.Through the ancient and Modern Medical Case Cloud Platform V 2.3.7,the drug frequency,natures,flavors,and channel tropism were analyzed.The cluster analysis,correlation analysis,complex network analysis and the core prescription were carried out.Eight SPF 6-8 week old SD male rats were divided into Chinese medicine group and control group by random number table method,with four rats in each group.The Chinese medicine group and control group were gavaged with 2 ml of core prescription herbal decoction and the same dose of saline for 7 d,respectively,and the drug-containing serum was extracted.CCK8 method was used to determine the optimal intervention concentration and intervention time of drug-containing serum on A549 and HepG2 cells.Blank groups were set up respectively:complete medium culture,no drug;Chinese medicine group:intervention with the best concentration of drug-containing serum;control group:intervention with the same dose of blank serum.Transwell assay was used to detect cell migration and invasion ability.Flow cytometry was used to detect apoptosis and cell cycle.Western blot was used to detect the expression of apoptosis and epithelial mesenchymal transition related proteins.Results A total of 8166 prescriptions were included,with a total of 300 kinds of drugs.The medicinal properties were warm and sweet,and the liver and lung meridians were the main meridians.The frequency of use of 16 drugs such as Radix Astragali,Carapax Trionycis,Mume Fructus,and cornus were>90%.Combining the results of cluster analysis and complex network analysis with the experience of Professor Li Zhong,one core prescription of the pingtong yin-yang method was derived,including raw Radix Astragali,roasted Radix Astragali,Carapax Trionycis shell,Agrimoniae Herba,Mume Fructus,Codonopsis Radix,Angelicae Sinensis Radix,Rehmanniae Radix Praeparata,Lonicerae Japonicae Caulis,Paeoniae Radix Alba,Dioscoreae Nipponicae Rhizoma,Fritillariae Thunbergii Bulbus,Bombyx Batryticatus,raw Concha Ostreae,and cornus.It was found that the optimal intervention concentration of the herbal medicine containing serum was 20%and the duration of the intervention was 48 h.The migration and invasion ability of A549 and HepG2 cells in the Chinese medicine group was lower than that of the control group,the apoptosis rate was higher than that of the control group,the blocked cell cycle was longer than that of the control group,and the expression levels of Bcl-2,N-Cadherin,and Vimentin were lower than those of the control group,and the expression levels of Cleaved-Caspase-3 and E-Cadherin were higher than those of the control group(P<0.05).Conclusion By using data mining,the core prescription can be obtained.The prescription drug-containing serum can inhibit the proliferation,migration,and invasion of tumor cells.The mechanism may be related to promoting apoptosis,blocking cell cycle,and inhibiting epithelial-mesenchymal transition process.
作者
孙可意
李忠
SUN Keyi;LI Zhong(Department of Hematology and Oncology,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing100700,China)
出处
《中国医药导报》
CAS
2024年第20期8-16,共9页
China Medical Herald
基金
北京市科技计划课题(Z161100001816014)。
关键词
厥阴
恶性肿瘤
核心处方
阴阳理论
数据挖掘
机制
Jueyin
Malignant cancer
Core prescription
Yin and yang theory
Data mining
Mechanism
