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Investigation and experimental validation of curcumin-related mechanisms against hepatocellular carcinoma based on network pharmacology

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2022年第8期23卷 682-698页

作者： Yang CHEN Qian LI Sisi REN Ting CHEN bingtao zhai Jiangxue CHENG Xiaoyan SHI Liang SONG Yu FAN Dongyan GUO College of Basic Medicine Shaanxi University of Chinese MedicineXianyang 712046China School of Pharmacy Shaanxi University of Chinese MedicineXianyang 712046China Medical Experimental Center Shaanxi University of Chinese MedicineXianyang 712046China Shaanxi Provincial Key Laboratory of TCM Constitution and Disease Prevention Shaanxi University of Chinese MedicineXianyang 712046China

Objective:To determine the potential molecular mechanisms underlying the therapeutic effect of curcumin on hepatocellular carcinoma(HCC)by network pharmacology and experimental in vitro validation.Methods:The predictive targets of curcumin or HCC were collected from several databases.the identified overlapping targets were crossed with Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses using the Database for Annotation,Visualization,and Integrated Discovery(DAVID)platform.Two of the candidate pathways were selected to conduct an experimental verification.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium(MTT)assay was used to determine the effect of curcumin on the viability of Hep G2 and LO2 cells.The apoptosis and autophagy of Hep G2 cells were respectively detected by flow cytometry and transmission electron microscopy.Besides,western blot and real-time polymerase chain reaction(PCR)were employed to verify the p53 apoptotic pathway and adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)autophagy pathway.Hep G2 cells were pretreated with pifithrin-α(PFT-α)and GSK690693 for further investigation.Results:The 167 pathways analyzed by KEGG included apoptosis,autophagy,p53,and AMPK pathways.The GO enrichment analysis demonstrated that curcumin was involved in cellular response to drug,regulation of apoptotic pathway,and so on.The in vitro experiments also confirmed that curcumin can inhibit the growth of Hep G2 cells by promoting the apoptosis of p53 pathway and autophagy through the AMPK pathway.Furthermore,the protein and messenger RNA(m RNA)of the two pathways were downregulated in the inhibitor-pretreated group compared with the experimental group.The damage-regulated autophagy modulator(DRAM)in the PFT-α-pretreated group was downregulated,and p62 in the GSK690693-pretreated group was upregulated.Conclusions:Curcumin can treat HCC through the p53 apoptotic pathway and the AMPK/Unc-51-like kinase 1(ULK1)autophagy pathway,in which the mutual transformati

关键词： Curcumin Network pharmacology p53 Adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK) Apoptosis Autophagy

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羌活胜湿汤物质基准量值传递分析

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中国药学杂志 2024年第2期59卷 128-135页

作者：祁庆瑞翟秉涛岳宝森赵锋田欢陕西中医药大学陕西省中药基础与新药研究重点实验室西安712046 西安市中医医院药剂科西安710021

目的建立羌活胜湿汤(Qianghuo Shengshi decoction, QSD)物质基准的指纹图谱及多指标成分含量测定方法,研究QSD物质基准量值传递规律,为QSD后续开发及质量控制评价提供依据。方法依据古籍记载制备15批QSD物质基准样品溶液,测定其出膏... 详细信息

目的建立羌活胜湿汤(Qianghuo Shengshi decoction, QSD)物质基准的指纹图谱及多指标成分含量测定方法,研究QSD物质基准量值传递规律,为QSD后续开发及质量控制评价提供依据。方法依据古籍记载制备15批QSD物质基准样品溶液,测定其出膏率,建立指纹图谱并进行相似度评价,明确特征峰及特征峰归属,完成QSD物质基准中8个指标成分的含量测定及饮片-物质基准量值传递分析。结果 15批QSD物质基准指纹图谱相似度处于0.882~0.938之间,共确定24个共有峰,并指认出8个共有峰分别为绿原酸、升麻素苷、阿魏酸、洋川芎内脂Ⅰ、5-O-甲基维斯阿米醇苷、甘草酸铵、羌活醇、蛇床子素。15批QSD物质基准的出膏率为22.54%~29.92%,平均出膏转移率为70.69%。各指标性成分的含量及饮片-基准样品的平均转移率分别为绿原酸0.077 2%~0.106 6%和128.05%;升麻素苷0.015 5%~0.023 8%和120.26%;阿魏酸0.047 5%~0.076 0%和114.49%;洋川芎内脂Ⅰ0.006 6%~0.008 9%和132.93%;5-O-甲基维斯阿米醇苷0.007 2%~0.009 9%和90.27%;甘草酸铵0.181 6%~0.273 5%和80.53%;羌活醇0.000 7%~0.001 3%和50.97%;蛇床子素0.000 9%~0.002 2%和13.67%。结论本实验采用指纹图谱、出膏率及多指标成分含量测定相结合的方法,对QSD饮片-物质基准的全过程进行量值传递分析,为该方的质量控制及复方制剂的开发提供参考。

关键词：羌活胜湿汤物质基准指纹图谱量值传递

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Preparation, characterization, pharmacokinetics and anticancer effects of PEGylated β-elemene liposomes

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Cancer Biology & Medicine 2020年第1期17卷 60-75页

作者： bingtao zhai Qibiao Wu Wengang Wang Mingming Zhang Xuemeng Han Qiujie Li Peng Chen Xiaying Chen Xingxing Huang Guohua Li Qin Zhang Ruonan Zhang Yu Xiang Shuiping Liu Ting Duan Jianshu Lou Tian Xie Xinbing Sui College of Pharmacy Chengdu University of Traditional Chinese MedicineChengdu 611137China Department of Holistic Integrative Pharmacy Institutes and Comprehensive Cancer Diagnosis and Treatment Center the Affiliated Hospital of Hangzhou Normal UniversityCollege of MedicineHangzhou Normal UniversityHangzhou 310018China State Key Laboratory of Quality Research in Chinese Medicines Faculty of Chinese MedicineMacao University of Science and TechnologyMacao 519020China Key Laboratory of Elemene Class Anticancer Chinese Medicine of Zhejiang Province and Engineering Laboratory of Development and Application of Traditional Chinese Medicine from Zhejiang Province Hangzhou Normal UniversityHangzhou 310018China

Objective:This study aimed to develop a new polyethylene glycol(PEG)ylatedβ-elemene liposome(PEG-Lipo-β-E)and evaluate its characterization,pharmacokinetics,antitumor effects and safety in vitro and in vivo.Methods:The liposomes were prepared by ethanol injection and high-pressure micro-jet homogenization.Characterization of the liposomes was conducted,and drug content,entrapment efficiency(EE),in vitro release and stability were studied by ultra-fast liquid chromatography(UFLC)and a liquid surface method.Blood was drawn from rats to establish the pharmacokinetic parameters.The anticancer effect was evaluated in a KU-19-19 bladder cancer xenograft model.Histological analyses were performed to evaluate safety.Results:The PEG-Lipo-β-E showed good stability and was characterized as 83.31±0.181 nm in size,0.279±0.004 in polydispersity index(PDI),-21.4±1.06 mV in zeta potential,6.65±0.02 in pH,5.024±0.107 mg/mL inβ-elemene(β-E)content,and 95.53±1.712%in average EE.The Fourier transform infrared spectroscopy(FTIR)and differential scanning calorimetry(DSC)indicated the formation of PEG-Lipo-β-E.Compared to elemene injection,PEG-Lipo-β-E demonstrated a 1.75-fold decrease in clearance,a 1.62-fold increase in half-life,and a 1.76-fold increase in area under the concentration-time curves(AUCs)from 0 hour to 1.5 hours(P<0.05).PEG-Lipo-β-E also showed an enhanced anticancer effect in vivo.Histological analyses showed that there was no evidence of toxicity to the heart,kidney,liver,lung or spleen.Conclusions:The present study demonstrates PEG-Lipo-β-E as a new formulation with ease of preparation,high EE,good stability,improved bioavailability and antitumor effects.

关键词： β-elemene PEGylated liposome pharmacokinetics antitumor effect bladder cancer

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An ATF24 peptide-functionalized β-elemene-nanostructured lipid carrier combined with cisplatin for bladder cancer treatment

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Cancer Biology & Medicine 2020年第3期17卷 676-692页

作者： bingtao zhai Peng Chen Wengang Wang Shuiping Liu Jiao Feng Ting Duan Yu Xiang Ruonan Zhang Mingming Zhang Xuemeng Han Xiaying Chen Qiujie Li Guohua Li Ying Liu Xingxing Huang Wenzheng Zhang Ting Pan Lili Yan Ting Jin Tian Xie Xinbing Sui Department of Medical Oncology the Affiliated Hospital of Hangzhou Normal UniversityCollege of MedicineHangzhou Normal UniversityHangzhou 310018China Key Laboratory of Elemene Class Anti-cancer Chinese Medicine of Zhejiang Provinee and Engineering Laboratory of Development and Application of Traditional Chinese Medicine from Zhejiang Province Hangzhou Normal UniversityHangzhou 310018China College of Pharmacy Chengdu University of Traditional Chinese MedicineChengdu 611137China Department of Medical Oncology Sir Run Run Shaw HospitalZhejiang UniversityHangzhou 310016China

Objective:In this study,we aimed to develop an amino-terminal fragment(ATF)peptide-targeted liposome carryingβ-elemene(ATF24-PEG-Lipo-β-E)for targeted delivery into urokinase plasminogen activator receptor-overexpressing bladder cancer cells combined with cisplatin(DDP)for bladder cancer treatment.Methods:The liposomes were prepared by ethanol injection and high-pressure microjet homogenization.The liposomes were characterized,and the drug content,entrapment efficiency,andin vitro release were studied.The targeting efficiency was investigated using confocal microscopy,ultra-fast liquid chromatography,and an orthotopic bladder cancer model.The effects of ATF24-PEG-Lipo-β-E combined with DDP on cell viability and proliferation were evaluated by a Cell Counting Kit-8(CCK-8)assay,a colony formation assay,and cell apoptosis and cell cycle analyses.The anticancer effects were evaluated in a KU-19-19 bladder cancer xenograft model.Results:ATF24-PEG-Lipo-β-E had small and uniform sizes(~79 nm),high drug loading capacity(~5.24 mg/mL),high entrapment efficiency(98.37±0.95%),and exhibited sustained drug release behavior.ATF24-PEG-Lipo-β-E had better targeting efficiency and higher cytotoxicity than polyethylene glycol(PEG)ylatedβ-elemene liposomes(PEG-Lipo-β-E).DDP,combined with ATF24-PEG-Lipo-β-E,exerted a synergistic effect on cellular apoptosis and cell arrest at the G2/M phase,and these effects were dependent on the caspase-dependent pathway and Cdc25C/Cdc2/cyclin B1 pathways.Furthermore,thein vivo antitumor activity showed that the targeted liposomes effectively inhibited the growth of tumors,using the combined strategy.Conclusions:The present study provided an effective strategy for the targeted delivery ofβ-elemene(β-E)to bladder cancer,and a combined strategy for bladder cancer treatment.

关键词： uPAR β-elemene active targeting liposome bladder cancer DDP

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基于转录组测序探讨金天格胶囊调控滑膜间充质干细胞外泌体miRNA和关节软骨细胞mRNA治疗大鼠骨关节炎的作用机制

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中国现代应用药学 2024年第11期41卷 1464-1475页

作者：王智超张雪张小飞史亚军郭东艳栾飞翟秉涛邹俊波袁普卫陕西中医药大学药学院陕西省中药基础与新药研究重点实验室陕西咸阳712046 陕西中医药大学附属医院陕西咸阳712046 陕西中医药大学第一临床医学院陕西咸阳712046

目的对金天格胶囊治疗骨关节炎的药效进行确证,基于高通量测序技术探讨滑膜间充质干细胞外泌体(synovial mesenchymal stem cell exosomes,SMSC-Exos)与关节软骨细胞(articular chondrocytes,ACs)治疗骨关节炎的潜在作用机制。方法采用... 详细信息

目的对金天格胶囊治疗骨关节炎的药效进行确证,基于高通量测序技术探讨滑膜间充质干细胞外泌体(synovial mesenchymal stem cell exosomes,SMSC-Exos)与关节软骨细胞(articular chondrocytes,ACs)治疗骨关节炎的潜在作用机制。方法采用Ⅱ型胶原酶诱导大鼠骨关节炎,通过一般行为学观察、双足平衡差异试验、机械足反射阈值、MicroCT观察与番红固绿染色进行药效验证。将SMSC、ACs在适宜浓度的含药血清中培养,对照组、模型组和金天格组ACs进行mRNA测序以及SMSC-Exos进行miRNA测序。筛选差异表达mRNA与差异表达miRNA,并进行靶基因预测。通过Venn图对SMSC与ACs的差异表达基因求交集得到共同差异表达基因,运用Cytoscape软件构建miRNA-m RNA调控网络。对共同差异表达基因进行表达趋势分析以及对差异表达基因mRNA与miRNA、Micro-CT药效指标与差异表达基因mRNA进行相关性分析。结果骨关节炎病理状态下,miRNA-23a-3p、miRNA-342-3p、miRNA-146b-5p、mi RNA-501-3p、miRNA-214-3p表达下调,miRNA-222-3p、miRNA-30e-3p、miRNA-676-3p、miRNA-192-5p表达上调(P<0.05),这些miRNA的表达均在金天格胶囊含药血清干预后逆转表达,且趋势明显。Micro-CT药效指标、m RNA与miRNA之间存在一定的相关性。结论金天格胶囊治疗骨关节炎的药效作用明显,其作用机制可能与促进SMSC-Exos靶向ACs转运miRNA进而调控Serpinb10、Ntn1、Il1b、Tgm2、Megf10、Il11、Cd40、Slc15a3、Pou2f2等基因有关。

关键词：骨关节炎转录组测序金天格胶囊滑膜间充质干细胞外泌体关节软骨细胞

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Erianin,a novel dibenzyl compound in Dendrobium extract,inhibits lung cancer cell growth and migration via calcium/calmodulin-dependent ferroptosis

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Signal Transduction and Targeted Therapy 2020年第1期5卷 1882-1892页

作者： Peng Chen Qibiao Wu Jiao Feng Lili Yan Yitian Sun Shuiping Liu Yu Xiang Mingming Zhang Ting Pan Xiaying Chen Ting Duan Lijuan zhai bingtao zhai Wengang Wang Ruonan Zhang Bi Chen Xuemeng Han Yicong Li Liuxi Chen Ying Liu Xingxing Huang Ting Jin Wenzheng Zhang Hong Luo Xiaohui Chen Yongqiang Li Qiujie Li Guohua Li Qin Zhang Lvjia Zhuo Zuyi Yang Huifen Tang Tian Xie Xiaoping Ouyang Xinbing Sui Holistic Integrative Pharmacy Institutes and Department of Medical Oncology the Affiliated Hospital of Hangzhou Normal UniversityCollege of MedicineHangzhou Normal UniversityHangzhouZhejiangChina Institute of Chinese Materia Medica Shanghai University of Traditional Chinese MedicineShanghai 201203China Key Laboratory of Elemene Class Anti-cancer Chinese Medicine of Zhejiang Province Hangzhou Normal UniversityHangzhouZhejiangChina State Key Laboratory of Quality Research in Chinese Medicines Faculty of Chinese MedicineMacao University of Science and TechnologyMacaoP.R.China Zhejiang Chinese Medicinal University HangzhouChina Dongzhimen Hospital Beijing University of Chinese MedicineBeijingChina Department of Medical Oncology Zhejiang UniversityHangzhouChina Department of Hematology and Oncology the Affiliated Hospital of Hangzhou Normal UniversityCollege of MedicineHangzhou Normal UniversityHangzhouZhejiangChina Radiation Detection Research Center Northwest Institute of Nuclear TechnologyXi’anChina Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica School of PharmacyNanjing University of Chinese MedicineNanjingChina

Ferroptosis,a novel form of programmed cell death,is characterized by iron-dependent lipid peroxidation and has been shown to be involved in multiple diseases,including cancer.Stimulating ferroptosis in cancer cells may be a potential strategy for cancer therapy.Therefore,ferroptosis-inducing drugs are attracting more attention for cancer treatment.Here,we showed that erianin,a natural product isolated from Dendrobium chrysotoxum Lindl,exerted its anticancer activity by inducing cell death and inhibiting cell migration in lung cancer cells.Subsequently,we demonstrated for the first time that erianin induced ferroptotic cell death in lung cancer cells,which was accompanied by ROS accumulation,lipid peroxidation,and GSH depletion.The ferroptosis inhibitors Fer-1 and Lip-1 but not Z-VAD-FMK,CQ,or necrostatin-1 rescued erianin-induced cell death,indicating that ferroptosis contributed to erianin-induced cell death.Furthermore,we demonstrated that Ca^(2+)/CaM signaling was a critical mediator of erianin-induced ferroptosis and that blockade of this signaling significantly rescued cell death induced by erianin treatment by suppressing ferroptosis.Taken together,our data suggest that the natural product erianin exerts its anticancer effects by inducing Ca^(2+)/CaMdependent ferroptosis and inhibiting cell migration,and erianin will hopefully serve as a prospective compound for lung cancer treatment.

关键词： peroxidation lung drugs

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PCDH17 increases the sensitivity of colorectal cancer to 5-fluorouracil treatment by inducing apoptosis and autophagic cell death

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Signal Transduction and Targeted Therapy 2019年第1期4卷 176-185页

作者： Shuiping Liu Haoming Lin Da Wang Qiang Li Hong Luo Guoxiong Li Xiaohui Chen Yongqiang Li Peng Chen bingtao zhai Wengang Wang Ruonan Zhang Bi Chen Mingming Zhang Xuemeng Han Qiujie Li Liuxi Chen Ying Liu Xiaying Chen Guohua Li Yu Xiang Ting Duan Jiao Feng Jianshu Lou Xingxing Huang Qin Zhang Ting Pan Lili Yan Ting Jin Wenzheng Zhang Lvjia Zhuo Yitian Sun Tian Xie Xinbing Sui Holistic Integrative Pharmacy Institutes and Comprehensive Cancer Diagnosis and Treatment Center the Affiliated Hospital of Hangzhou Normal UniversityCollege of MedicineHangzhou Normal UniversityHangzhouZhejiangChina Key Laboratory of Elemene Class Anti-cancer Chinese Medicine of Zhejiang Province and Engineering Laboratory of Development and Application of Traditional Chinese Medicine from Zhejiang Province Hangzhou Normal UniversityHangzhouZhejiangChina Department of Hepatobiliary Pancreatic Surgery Sun Yat-sen Memorial HospitalSun Yat-sen UniversityGuangzhouChina Department of Colorectal Surgery The Second Affiliated Hospital of Zhejiang University School of MedicineHangzhouChina Department of Ultrasound Medicine The First Affiliated HospitalSchool of MedicineZhejiang UniversityHangzhouChina Department of Medical Oncology Sir Run Run Shaw HospitalZhejiang UniversityHangzhouChina Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica School of PharmacyNanjing University of Chinese MedicineNanjingChina

5-Fluorouracil(5-FU)is known as a first-line chemotherapeutic agent against colorectal cancer(CRC),but drug resistance occurs frequently and significantly limits its clinical success.Our previous study showed that the protocadherin 17(PCDH17)gene was frequently methylated and functioned as a tumor suppressor in CRC.However,the relationship between PCDH17 and 5-FU resistance in CRC remains unclear.Here,we revealed that PCDH17 was more highly expressed in 5-FU-sensitive CRC tissues than in 5-FU-resistant CRC tissues,and high expression of PCDH17 was correlated with high BECN1 expression.Moreover,this expression profile contributed to superior prognosis and increased survival in CRC patients.Restoring PCDH17 expression augmented the 5-FU sensitivity of CRC in vitro and in vivo by promoting apoptosis and autophagic cell death.Furthermore,autophagy played a dominant role in PCDH17-induced cell death,as an autophagy inhibitor blocked cell death to a greater extent than the pancaspase inhibitor Z-VAD-FMK.PCDH17 inhibition by siRNA decreased the autophagy response and 5-FU sensitivity.Mechanistically,we showed that c-Jun NH2-terminal kinase(JNK)activation was a key determinant in PCDH17-induced autophagy.The compound SP600125,an inhibitor of JNK,suppressed autophagy and 5-FU-induced cell death in PCDH17-reexpressing CRC cells.Taken together,our findings suggest for the first time that PCDH17 increases the sensitivity of CRC to 5-FU treatment by inducing apoptosis and JNK-dependent autophagic cell death.PCDH17 may be a potential prognostic marker for predicting 5-FU sensitivity in CRC patients.

关键词： death treatment apoptosis

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宣肺止咳颗粒的制备工艺及薄层色谱鉴别

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陕西中医药大学学报 2024年第4期47卷 78-86页

作者：王智超石长楠史亚军栾飞张小飞郭东艳翟秉涛雷根平邹俊波陕西省中药基础与新药研究重点实验室/陕西中医药大学药学院陕西咸阳712046 西安恒生堂制药有限公司陕西西安710000 陕西中医药大学附属医院陕西咸阳712046

目的筛选宣肺止咳颗粒剂制备成型稳定可行的最优工艺,为其质量研究奠定基础。方法选用橙皮苷含量和干膏率的不同权重比为综合评价指标,进行单因素实验,发现提取次数、时间及处方加水倍数对实验结果影响显著,故对饮片粉碎粒度、浸泡时... 详细信息

目的筛选宣肺止咳颗粒剂制备成型稳定可行的最优工艺,为其质量研究奠定基础。方法选用橙皮苷含量和干膏率的不同权重比为综合评价指标,进行单因素实验,发现提取次数、时间及处方加水倍数对实验结果影响显著,故对饮片粉碎粒度、浸泡时间不予考虑,以此为基础进行正交实验,最终优选出宣肺止咳颗粒的水提工艺。考察药液比、高速离心的转速和时间,筛选出较优的药液除杂工艺;将常压浓缩与减压浓缩药液的橙皮苷峰面积、干膏率进行比较,确定浓缩工艺;选用减压干燥方法进行干燥,并确定该方法的可行性;选用不同浓度乙醇,将玉米淀粉和糊精(1∶1)、蔗糖和糊精(1∶1)与干膏粉分别混合进行制粒,对比制得颗粒的性状,筛选出成型工艺。根据2020版《中国药典》一部,对宣肺止咳方中的瓜蒌皮、清半夏两味药材进行定性鉴别,并对宣肺止咳颗粒橙皮苷含量测定的方法建立质量标准。结果全方加10倍量水,提取2次,每次60 min;浓缩至药液比为1∶3,选用高速离心法分离纯化,固定转速4000 r·min^(-1),时间10 min;在80℃下进行减压浓缩,结果最优;减压干燥方法稳定可靠,适用于本方;优选出糊精和玉米淀粉(1∶1)与药膏干粉1∶1混合,喷洒90%乙醇,制得的颗粒性质最好;宣肺止咳颗粒橙皮苷含量测定方法稳定可行;薄层鉴别方法简便可靠。结论筛选出的宣肺止咳颗粒的制备工艺稳定可靠、科学合理,可应用于企业化大生产。

关键词：宣肺止咳颗粒橙皮苷正交实验 HPLC 薄层色谱

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葛根-南五味子预防给药减轻四氯化碳致小鼠急性肝损伤

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中药药理与临床 2023年第11期39卷 39-45页

作者：郭伟胡田颖雷宏强詹博李欢翟秉涛郭东艳渭南市中心医院渭南714000 陕西中医药大学秦药特色资源研究与开发国家重点实验室(培育)陕西省中药基础与新药研究重点实验室咸阳712046

目的:研究葛根-南五味子对四氯化碳(CCl4)诱导的急性肝损伤的保护作用,并初步探究其作用机制。方法:48只c57BL/6雄性小鼠随机分为正常对照组、模型对照组、水飞蓟素0.2 g/kg组、葛根-南五味子2.5、5、10 g/kg组,每组8只,连续给药14 d。... 详细信息

目的:研究葛根-南五味子对四氯化碳(CCl4)诱导的急性肝损伤的保护作用,并初步探究其作用机制。方法:48只c57BL/6雄性小鼠随机分为正常对照组、模型对照组、水飞蓟素0.2 g/kg组、葛根-南五味子2.5、5、10 g/kg组,每组8只,连续给药14 d。末次给药后,除正常对照组外,每只小鼠按10 mL/kg腹腔注射2%CCl4橄榄油溶液,禁食16 h后收集小鼠血清和肝脏。生化法检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆固醇(TC)、三酰甘油(TG)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)和还原性谷胱甘肽(GSH)的活力或含量;ELISA法检测血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、IL-1β和IL-10的含量;TUNEL法检测小鼠肝组织细胞凋亡率;免疫荧光检测F4/80阳性率情况;免疫组化法检测肝组织中细胞色素P4502E1(CYP2E1)、中性粒细胞表面抗原(Ly6G)、BCL2-Associated X(BAX)、B淋巴细胞瘤-2基因(BCL-2)和半胱氨酸蛋白酶-3(Caspase-3)的蛋白表达。结果:与正常对照组比较,模型对照组出现肝细胞气球样变、脂变和坏死以及炎性浸润;血清中ALT、AST、TC、TG、TNF-α、IL-6和IL-1β活力或含量显著升高,且IL-10含量显著降低(P<0.01);肝组织中SOD活力、CAT和GSH含量显著降低,且MDA含量显著升高(P<0.01);肝组织中细胞凋亡率和F4/80阳性率显著增加(P<0.01);肝组织中CYP2E1、Ly6G、BAX、Caspase-3蛋白阳性表达显著增加,且BCL-2蛋白阳性表达显著降低(P<0.01);与模型对照组比较,葛根-南五味子各组呈剂量依赖性地缓解肝细胞气球样变、脂变和坏死减少以及炎症浸润,降低血清中ALT、AST、TC、TG、TNF-α、IL-6和IL-1β活力或含量,增加IL-10含量,增加肝组织中SOD、CAT和GSH活力或含量,降低MDA含量;减少肝组织中细胞凋亡率和F4/80阳性率,降低肝组织中CYP2E1、Ly6G、BAX、Caspase-3蛋白阳性表达,升高BCL-2蛋白阳性表达。结论:葛根-南五味子预防给药对CCl4所致肝损伤小鼠具有一定保护作用,其机制可能与增强抗氧化能力、抑制炎症和细胞凋亡有关。

关键词：葛根-南五味子急性肝损伤氧化应激炎症细胞凋亡保肝作用

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基于网络药理学和Box-Behnken响应面法优化养心颗粒提取工艺

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中国现代应用药学 2023年第15期40卷 2064-2075页

作者：王嘉雯王洁樊强强翟秉涛张小飞程江雪郭东艳陕西中医药大学秦药特色资源研究与开发国家重点实验室(培育)陕西省中药基础与新药研究重点实验室西安712046 陕西中医药大学附属医院西安712046

目的通过网络药理学和Box-Behnken响应面法优化养心颗粒提取工艺。方法通过网络药理学挖掘养心颗粒活性成分并初步探究其治疗冠心病的作用机制;分子对接技术预测活性成分与主要靶点的结合能力。同时结合该方君臣佐使配伍关系、成分药... 详细信息

目的通过网络药理学和Box-Behnken响应面法优化养心颗粒提取工艺。方法通过网络药理学挖掘养心颗粒活性成分并初步探究其治疗冠心病的作用机制;分子对接技术预测活性成分与主要靶点的结合能力。同时结合该方君臣佐使配伍关系、成分药理作用及中国药典2020年版一部各药味含量测定指标成分,进一步确定养心颗粒工艺评价指标成分。此外,结合层次分析法确定各成分权重,Box-Behnken响应面法优化养心颗粒提取工艺。结果筛选得到的养心颗粒主要活性成分为梓醇、地黄苷D、毛蕊花糖苷、阿魏酸和党参炔苷,通过作用于AKT1、IL-6、IL-1β、VEGFA、JUN、MAPK3等核心靶标,调节脂质和动脉粥样硬化、MAPK信号等通路共同发挥治疗冠心病的作用。分子对接结果表明,网络药理学预测的活性成分与核心靶点结合能均<–5.0 kJ·mol^(–1)。层次分析法计算得到5种成分梓醇、地黄苷D、毛蕊花糖苷、阿魏酸、党参炔苷权重系数分别为0.329 7,0.329 7,0.164 8,0.109 9,0.065 9,Box-Behnken响应面法得到最优水提工艺:采用10倍量的水提取2次,每次煎煮1.5 h。验证试验表明5种成分含量与预测值相符,RSD值均<5%。结论基于网络药理学和Box-Behnken响应面法得到的养心颗粒提取工艺稳定可行,为其进一步质量提升提供了实验依据。

关键词：养心颗粒网络药理学层次分析法 Box-Behnken响应面法提取工艺

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