BACKGROUND:The phosphoinositide 3-kinase/protein kinase-B/mechanistic target of rapamycin (PI3K/Akt/mTOR) signalling pathway is crucial for cell survival, differentiation, apoptosis and metabolism. Xihuang pills (XHP) are a traditional Chinese preparation with antitumour properties. They inhibit the growth of breast cancer, glioma, and other tumours by regulating the PI3K/Akt/mTOR signalling pathway. However, the effects and mechanisms of action of XHP in hepatocellular carcinoma (HCC) remain unclear. Regulation of the PI3K/Akt/mTOR signalling pathway effectively inhibits the progression of HCC. However, no study has focused on the XHP-associated PI3K/Akt/mTOR signalling pathway. Therefore, we hypothesized that XHP might play a role in inhibiting HCC through the PI3K/Akt/mTOR signalling pathway. AIM:To confirm the effect of XHP on HCC and the possible mechanisms involved. METHODS:The chemical constituents and active components of XHP were analysed using ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). Cell-based experiments and in vivo xenograft tumour experiments were utilized to evaluate the effect of XHP on HCC tumorigenesis. First, SMMC-7721 cells were incubated with different concentrations of XHP (0, 0.3125, 0.625, 1.25, and 2.5 mg/mL) for 12 h, 24 h and 48 h. Cell viability was assessed using the CCK-8 assay, followed by an assessment of cell migration using a wound healing assay. Second, the effect of XHP on the apoptosis of SMMC-7721 cells was evaluated. SMMC-7721 cells were stained with fluorescein isothiocyanate and annexin V/propidium iodide. The number of apoptotic cells and cell cycle distribution were measured using flow cytometry. The cleaved protein and mRNA expression levels of caspase-3 and caspase-9 were detected using Western blotting and quantitative reverse-transcription polymerase chain reaction (RT-qPCR), respectively. Third, Western blotting and RT-qPCR were performed to confirm the effects of XHP on the protein and mRNA expression of components of the PI3K/Akt/mTOR signalling pathway. Finally, the effects of XHP on the tumorigenesis of subcutaneous hepatocellular tumours in nude mice were assessed. RESULTS:The following 12 compounds were identified in XHP using high-resolution mass spectrometry: Valine, 4-gingerol, myrrhone, ricinoleic acid, glycocholic acid, curzerenone, 11-keto-β-boswellic acid, oleic acid, germacrone, 3-acetyl-9,11-dehydro-β-boswellic acid, 5β-androstane-3,17-dione, and 3-acetyl-11-keto-β-boswellic acid. The cell viability assay results showed that treatment with 0.625 mg/mL XHP extract decreased HCC cell viability after 12 h, and the effects were dose- and time-dependent. The results of the cell scratch assay showed that the migration of HCC cells was significantly inhibited in a time-dependent manner by the administration of XHP extract (0.625 mg/mL). Moreover, XHP significantly inhibited cell migration and resulted in cell cycle arrest and apoptosis. Furthermore, XHP downregulated the PI3K/Akt/mTOR signalling pathway, which activated apoptosis executioner proteins (e.g., caspase-9 and caspase-3). The inhibitory effects of XHP on HCC cell growth were determined in vivo by analysing the tumour xenograft volumes and weights. CONCLUSION:XHP inhibited HCC cell growth and migration by stimulating apoptosis via the downregulation of the PI3K/Akt/mTOR signalling pathway, followed by the activation of caspase-9 and caspase-3. Our findings clarified that the antitumour effects of XHP on HCC cells are mediated by the PI3K/Akt/mTOR signalling pathway, revealing that XHP may be a potential complementary therapy for HCC.

BACKGROUND:In traditional Chinese medicine (TCM), frankincense and myrrh are the main components of the antitumor drug Xihuang Pill. These compounds show anticancer activity in other biological systems. However, whether frankincense and/or myrrh can inhibit the occurrence of hepatocellular carcinoma (HCC) is unknown, and the potential molecular mechanism(s) has not yet been determined.AIM:To predict and determine latent anti-HCC therapeutic targets and molecular mechanisms of frankincense and myrrh in vivo.METHODS:In the present study, which was based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (http://tcmspw.com/tcmsp.php), Universal Protein database (http://www.uniprot.org), GeneCards: The Human Gene Database (http://www.genecards.org/) and Comparative Toxicogenomics Database (http://www.ctdbase.org/), the efficacy of and mechanism by which frankincense and myrrh act as anti-HCC compounds were predicted. The core prediction targets were screened by molecular docking. In vivo, SMMC-7721 human liver cancer cells were transplanted as xenografts into nude mice to establish a subcutaneous tumor model, and two doses of frankincense plus myrrh or one dose of an EGFR inhibitor was administered to these mice continuously for 14 d. The tumors were collected and evaluated: the tumor volume and growth rate were gauged to evaluate tumor growth; hematoxylin-eosin staining was performed to estimate histopathological changes; immunofluorescence (IF) was performed to detect the expression of CD31, α-SMA and collagen IV; transmission electron microscopy (TEM) was conducted to observe the morphological structure of vascular cells; enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of secreted HIF-1α and TNF-α; reverse transcription-polymerase chain reaction (RT-qPCR) was performed to measure the mRNA expression of HIF-1α, TNF-α, VEGF and MMP-9; and Western blot (WB) was performed to determine the levels of proteins expressed in the EGFR-mediated PI3K/Akt and MAPK signaling pathways.RESULTS:The results of the network pharmacology analysis showed that there were 35 active components in the frankincense and myrrh extracts targeting 151 key targets. The molecular docking analysis showed that both boswellic acid and stigmasterol showed strong affinity for the targets, with the greatest affinity for EGFR. Frankincense and myrrh treatment may play a role in the treatment of HCC by regulating hypoxia responses and vascular system-related pathological processes, such as cytokine-receptor binding, and pathways, such as those involving serine/threonine protein kinase complexes and MAPK, HIF-1 and ErbB signaling cascades. The animal experiment results were verified. First, we found that, through frankincense and/or myrrh treatment, the volume of subcutaneously transplanted HCC tumors was significantly reduced, and the pathological morphology was attenuated. Then, IF and TEM showed that frankincense and/or myrrh treatment reduced CD31 and collagen IV expression, increased the coverage of perivascular cells, tightened the connection between cells, and improved the shape of blood vessels. In addition, ELISA, RT-qPCR and WB analyses showed that frankincense and/or myrrh treatment inhibited the levels of hypoxia-inducible factors, inflammatory factors and angiogenesis-related factors, namely, HIF-1α, TNF-α, VEGF and MMP-9. Furthermore, mechanistic experiments illustrated that the effect of frankincense plus myrrh treatment was similar to that of an EGFR inhibitor with regard to controlling EGFR activation, thereby inhibiting the phosphorylation activity of its downstream targets: the PI3K/Akt and MAPK (ERK, p38 and JNK) pathways.CONCLUSION:In summary, frankincense and myrrh treatment targets tumor blood vessels to exert anti-HCC effects via EGFR-activated PI3K/Akt and MAPK signaling pathways, highlighting the potential of this dual TCM compound as an anti-HCC candidate.

Objective: To investigate the effects of Niuhuang (Bovis Calculus, BC) and Shexiang (Moschus) (BC-Moschus) on human hepatocellular carcinoma (HCC) cells SMMC-7721 and a nude mouse model of subcutaneous xenografts, and to explore its anti-HCC mechanism. Methods: The BC-Moschus combination was applied to two liver cancer models in vivo and in vitro. SMMC-7721 was divided into the BC-Moschus group and the control group, and different doses (rude drug dosage 0.625, 1.25, 2.5, and 5 mg/mL) of BC-Moschus extract were used for the intervention. The proliferation ability of HCC cells was detected using the Cell Counting Kit-8 (CCK-8) assay, and the migration ability was detected by a wound healing assay. A subcutaneous xenograft model was prepared using nude mice with human HCC. Specific pathogen-free-grade BALB/c nude mice (5-week-old) were randomly divided into the following groups (n = 6 per group): control (0.9% physiological saline 0.2 mL/d), BC-Moschus [BC 45.5 mg/(kg·d)+ Moschus 13 mg/(kg·d)], and cisplatin (DDP, intraperitoneal injection 5 mg/kg per week) groups. All groups were administered for 14 d. The volume and mass of the subcutaneous xenografts in nude mice were observed. The expression levels of phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway, apoptosis-associated factor p70 S6 Kinase (S6K), Bax, Bcl-2, caspase-3, and caspase-9 in nude mice subcutaneous xenografts were measured by real-time quantitative PCR (RT-qPCR) and Western blot. Terminal Deoxynucleotidy Transferase-Mediated dUTP Nick-End Labeling (TUNEL) was used for quantitative analysis of apoptotic cells. Results: The CCK-8 assay demonstrated that the BC-Moschus combination inhibited HCC cell proliferation in a superior manner to the use of BC and Moschus alone, and the inhibition effect was dose- and time-dependent (P < 0.01). The wound healing assay showed that the BC-Moschus combination inhibited HCC cell migration (P < 0.01). In the subcutaneous xenograft model of nude mice with human HCC, we found that the tumor volume and weight of the BC-Moschus group were lower than those of the control group (P < 0.01). The levels of the PI3K/AKT/mTOR signaling pathway and S6K protein in the BC-Moschus and DDP groups were significantly decreased (P < 0.01). The expression level of the anti-apoptotic gene Bcl-2 was downregulated (P < 0.05), and the expression of the pro-apoptotic gene Bax and apoptosis-related factors caspase-3 and caspase-9 were significantly upregulated (P < 0.01). The TUNEL assays further confirmed that the combination of the BC-Moschuas could promote HCC (P < 0.01). Conclusion: The BC-Moschus combination inhibited the proliferation and migration ability of HCC cells SMMC-7721 and effectively inhibited the growth of subcutaneous xenografts in nude mice. The mechanism may be closely related to the downregulation of the PI3K/AKT/mTOR pathway, regulation of apoptosis-related protein caspase-3, caspase-9, Bcl-2, and Bax expression, and promotion of apoptosis.

Gastrointestinal cancer may be associated with dysbiosis, which is characterized by an alteration of the gut microbiota. Understanding the role of gut microbiota in the development of gastrointestinal cancer is useful for cancer prevention and gut microbiota-based therapy. However, the potential role of dysbiosis in the onset of tumorigenesis is not fully understood. While accumulating evidence has demonstrated the presence of dysbiosis in the intestinal microbiota of both healthy individuals and patients with various digestive system diseases, severe dysbiosis is often present in patients with digestive system cancer. Importantly, specific bacteria have been isolated from the fecal samples of these patients. Thus, the association between dysbiosis and the development of digestive system cancer cannot be ignored. A new model describing this relationship must be established. In this review, we postulate that dysbiosis serves as the first hit for the development of digestive system cancer. Dysbiosis-induced alterations, including inflammation, aberrant immune response, bacteria-produced genotoxins, and cellular stress response associated with genetic, epigenetic, and/or neoplastic changes, are second hits that speed carcinogenesis. This review explains the mechanisms for these four pathways and discusses gut microbiota-based therapies. The content included in this review will shed light on gut microbiota-based strategies for cancer prevention and therapy.

鳖甲是临床常用药物,现代对鳖甲的研究主要聚焦于实验研究及临床观察,鲜见文献研究.该文在系统查阅鳖甲古今文献的基础上,通过分析历代本草方书中的相关内容,对鳖甲的名称、基原、产地、品质评价、功效主治、炮制方法及用药禁忌进行全面考证.通过考证发现,在基原上,古籍文献所载的鳖甲当来源于中华鳖Trionyx sinensis的背甲,山瑞鳖T.steindachneri的背甲不宜作为中药鳖甲的来源.鳖甲的道地产区在今长江中下游的岳阳、荆州、安徽东南部及江苏西部.关于鳖甲的品质评价,本草古籍中常以鳖甲的肋数如七肋、九肋作为品质评价的标准,但通过文献研究及对药材市场的实地考察,发现以肋数作为品质的评价标准在现代并不可取.随着时代的更迭,鳖甲的功效主治在《神农本草经》的基础上逐渐扩展,后世将鳖甲广泛应用于内外妇儿各科.需要注意的是,鳖甲治劳热骨蒸当来源于《神农本草经》,而不是《本草衍义》等古籍所言的《药性论》.在炮制方面,鳖甲炮制方法多样,主要以醋制为主.在用药禁忌方面,鳖甲不能与矾石、理石配伍,孕妇禁用,脾虚胃弱、肝虚无热者慎用.该文的考证结果为鳖甲的正本清源及进一步资源开发利用提供了参考依据.

目的:分析、总结经方中黄连"角药"的配伍特点与临床应用,并深入剖析以黄连为角药的类方鉴别,为现代临床用药谴方提供理论依据.方法:谨遵经旨,对经方中黄连"角药"的配伍特点与临床应用进行详尽梳理,并从独立成方之"角药"、作为方剂主要部分之"角药"以及类方鉴别三方面进行论述.结果:经方中独立成方之黄连"角药"两对、作为方剂主要部分之"角药"7 对,黄连的角药配伍,寒热并用,表里同调,虚实相兼,使得其可用于下利、呕吐、肠鸣、烦躁、心下痞等多个方证.结论:经方中蕴含着丰富的黄连"角药"配伍知识,理解掌握经方中黄连"角药"的配伍特点与临床应用及其类方的准确运用,可扩大主治,减少药味,精准治疗.

目的 运用网络药理学方法及分子对接技术对雷公藤治疗肝癌的主要活性成分及其潜在作用机制进行探讨.方法 通过中药系统药理学数据库与分析平台(TCMSP)筛选雷公藤主要活性成分及其预测靶点;在DrugBank、GeneCards、OMIM、TTD数据库中筛选肝癌相关靶点;通过R语言软件映射得到雷公藤治疗肝癌靶点,并绘制韦恩图;在STRING网站构建治疗靶点PPI网络图;应用Cytoscape软件构建"雷公藤-活性成分-靶点-肝癌"互作网络图;通过R语言软件对治疗靶点进行GO功能分析和KEGG通路富集分析;应用PubChem、RCSB PDB数据库,AutoDock、PyMOL软件对药物潜在活性成分与关键靶点进行分子对接.结果 共筛选出活性成分51个和潜在治疗靶点120个,靶点主要涉及酰胺结合、肽结合、DNA结合转录因子结合等生物学过程,并主要富集于卡波西肉瘤相关疱疹病毒感染、乙型肝炎、流体剪切应力和动脉粥样硬化等信号通路中.分子对接验证显示对接得分大于-5kcal/mol占100％,即所有靶点与成分的结合活性较好.结论 通过网络药理学方法及分子对接技术证实了雷公藤多成分、多靶点、多途径的作用特点,预测了雷公藤治疗肝癌的潜在作用机制,为后续进一步开发和应用提供理论依据.

目的 通过收集中医药治疗原发性肝癌经验病例,分析其组方规律,为临床治疗提供选方用药和思路借鉴.方法 计算机检索中国知网中医药治疗肝癌经验病例,以主题词为"原发性肝癌""中医""肝癌"检索中国知网自建库至2020年7月所收录文献,以药物使用频次、性味归经、及Apriori关联规则结果为观察指标进行统计分析.结果 纳入文献101篇,筛选出个113复方,涉及272味中药,词频最高的药物是白术、茯苓、柴胡、白花蛇舌草、黄芪、甘草、鸡内金、半枝莲、莪术、八月札、党参、麦芽等.使用药物在"四气"中所占比重由大到小依次为寒、温、平、凉、热.使用药物在五味中所占比重由大到小依次为甘、苦、辛、淡、酸、咸、涩.药物使用归经频次由多到少依次为脾、肝、胃、肺、心、肾、胆、大肠、小肠、膀胱、三焦、心包等.Apriori关联规则计算结果显示规则支持最高为茯苓+黄芪+白术配伍以及茯苓+白花蛇舌草+白术配伍.结论 中医治疗原发性肝癌以扶正健脾为组方首选,配伍疏肝、清热、化瘀软坚散结等药物.

肿瘤目前已成为危害人类生命健康的主要疾病之一.研究表明,龟鹿二仙胶可通过增强机体对肿瘤细胞的免疫、降低肿瘤细胞耐药性、改善骨髓抑制,从而发挥抗肿瘤作用,而成方中单药抗肿瘤作用亦可通过诱导细胞凋亡、抑制新生血管形成等多种途径来实现.虽然龟鹿二仙胶抗肿瘤机制研究已取得一定进展,但主要聚焦于免疫系统,今后需进一步研究其相关作用机制,为肿瘤临床治疗拓宽思路和方法.

目的 观察薯蓣丸对肿瘤微环境中HIF-1α与p53的表达以及线粒体损伤的影响,及其对肝细胞癌的治疗作用.方法 制备人肝癌裸鼠皮下移植瘤模型,雄性BALB/c裸鼠(n=24,5周龄),随机分为4组,每组6只:模型组(0.9％生理盐水0.2 mL/d)、薯蓣丸组(薯蓣丸0.4 g/d),薯蓣丸+顺铂组(薯蓣丸0.4 g/d+每周腹腔注射顺铂5 mg/kg)、顺铂组(每周腹腔注射顺铂5 mg/kg).每只裸鼠灌胃剂量为0.2 mL/d,每2天测量1次肿瘤体积与小鼠体质量,连续干预14 d后脱颈处死,剥离皮下移植瘤;电镜下观察肿瘤组织线粒体结构和数量;Western blot与RT-qPCR法分别检测人肝癌裸鼠皮下移植瘤中HIF-1α、p53的蛋白及mRNA表达.结果 与模型组相比,薯蓣丸组可抑制人肝癌裸鼠皮下移植瘤的生长,抑制HIF-1α的mRNA与蛋白表达(P<0.05),上调抑癌基因p53的m-RNA与蛋白表达(P<0.05),同时改善线粒体结构损伤;与薯蓣丸组或顺铂组相比,薯蓣丸联合顺铂组对HIF-1α、p53基因表达及改善线粒体结构作用更显著(P<0.05).结论 薯蓣丸可通过促使HIF-1α的失活与p53的活化,改善线粒体结构损伤,从而抑制人肝癌裸鼠皮下移植瘤的生长,与顺铂联用后具有协同增效作用.

目的 挖掘大柴胡汤治疗肝细胞癌的lncRNA-miRNA-mRNA调控网络,探讨其抗肝癌作用机制.方法 利用中药系统药理学数据库与分析平台(TCMSP)、GEO、miRDB、miRTarBase、TargetScan数据库获取大柴胡汤的活性成分、预测靶点及肝细胞癌的差异表达长链非编码RNA(lncRNAs)、微小RNA(miRNAs)和mRNAs,通过R语言和Perl软件筛选大柴胡汤治疗肝细胞癌的关键lncRNAs、miRNAs、mRNAs.采用Cytoscape3.7.0软件和STRING数据库构建活性成分-靶点网络、lncRNA-miRNA-mRNA转录网络和蛋白相互作用网络,进行GO和KEGG富集分析,并予分子对接验证,进行整合分析.结果 得到大柴胡汤治疗肝细胞癌的97个有效成分和42个靶点,构建了由19个关键lncRNAs、23个关键miRNAs和42个关键mRNAs组成的lncRNA-miRNA-mRNA转录网络.富集分析结果显示,这些靶基因参与了肿瘤的生存、凋亡、自噬、氧化应激、蛋白激酶等生物学过程,主要通过PI3K-Akt信号通路、MAPK信号通路和TNF信号通路等发挥作用.结论 大柴胡汤治疗肝细胞癌具有多成分、多靶点、多途径协同作用特点,可为相关研究提供依据和方向.

壁虎和石龙子药用历史悠久,但长期以来两者常混为一谈,而现代对两者的研究主要集中在临床观察、有效成分及抗癌机制等方面,鲜见文献研究.该文在系统查阅壁虎和石龙子古今文献的基础上,通过分析历代本草方书中的相关内容,对两者的名称、基原、功效主治、炮制方法及用药禁忌进行全面考证.通过考证发现,在先秦,壁虎与石龙子被当作同一物,至汉代则知两者为不同的动物,到三国时期两者又被混淆,直至明代《本草纲目》才明确两者的关系.在基原上,石龙子的基原包括蜥蜴科丽斑麻蜥Eremias argus,山地麻蜥E.brenchleyi及石龙子科中国石龙子Eumeses chinensis.壁虎的基原包括多疣壁虎Gekko japonicus,铅山壁虎G.hokouensis,无蹼壁虎G.swinhonis及蹼趾壁虎G.subpalmatus.功效主治方面,历代本草记载石龙子的主要功效为利水,壁虎的主要功效为祛风、化瘀.在炮制上壁虎与石龙子有多种炮制方法,但主要以传统"火制法"为主.关于用药禁忌,古籍文献记载石龙子恶硫黄、芜荑、斑蝥,孕妇忌用;壁虎慎用于血虚气弱之人.该文的考证结果厘清了壁虎和石龙子从名称、功效主治到炮制等多方面的历史沿革,为壁虎及石龙子的正本清源及进一步开发利用提供了理论依据.

蜈蚣作为中国传统中药材,因其走窜主力最速,有活血祛瘀、豁痰攻毒散结的功效,使其广泛被应用于临床的抗肿瘤治疗,尤其是肝癌的治疗领域.本文从蜈蚣入药的文献记载、药理机制及其在肝癌治疗中的临床应用等方面进行综述,为进一步研究提供借鉴及参考.

目的 分别研究大黄、灵芝、人参对蜈蚣提取液所致大鼠肝肾损伤的修复作用.方法 SD大鼠分为5组,正常组(蒸馏水)、蜈蚣组(4.5 g/kg)、蜈蚣(4.5 g/kg)+灵芝组(1.08 g/kg)、蜈蚣(4.5 g/kg)+人参组(0.8 g/kg)、蜈蚣(4.5 g/kg)+大黄组(0.45 g/kg),灌胃给药14 d,每日1次,观察大鼠的一般情况,检测各组谷丙转氨酶(Alanine transaminase,ALT)、血肌酐(Serum creatinine,SCr)、肝脏组织脂质过氧化物丙二醛(Malondialdehyde,MDA)活性及观察肝组织病理学改变.结果 正常组、蜈蚣+灵芝组、蜈蚣+人参组、蜈蚣+大黄组的谷丙转氨酶、血肌酐、丙二醛值均低于蜈蚣组,差异有统计学意义.灵芝、人参、大黄对蜈蚣所致肝组织的片状坏死,有不同程度的减轻.结论 灵芝、人参、大黄对蜈蚣所致肝肾损伤具有修复作用,其抗肝损伤作用机制可能与抗脂质过氧化有关.