甘草抗肿瘤作用研究进展

近年来,中医药在恶性肿瘤的治疗中发挥重要作用。甘草是最常用的中药之一,主要活性成分为三萜皂苷类、黄酮类以及香豆素类物质。研究发现,甘草可通过介导肿瘤细胞凋亡和自噬、抑制肿瘤细胞增殖和转移等途径发挥抗肿瘤作用。目前,甘草相关方剂,如大黄甘草汤、补中益气汤和六君子汤等,在肿瘤辅助治疗中多有应用,可缓解肿瘤疼痛、黏膜刺激、胃肠道不良反应、贫血等。针对甘草抗肿瘤的主要有效成分异甘草素水溶性差、生物利用度低、体内半衰期短的问题,纳米悬浮液、脂质-聚合物杂化纳米颗粒系统和聚合物胶束等新型药物递送系统的研究突飞猛进。开发甘草及其生物活性成分作为抗肿瘤药物具有巨大潜力和应用价值。     

甘草酸诱导人乳腺癌细胞凋亡和细胞内Ca2+的关系

引言 甘草酸（Glycyrrhizin,GL）是甘草中最重要的有效成分之一,有研究表明甘草酸对人癌细胞增殖有明显的抑制和诱导其凋亡的作用。本研究即探讨甘草酸诱导人乳腺癌细胞（MCF-7）凋亡与细胞内Ca^2＋浓度变化的关系。     

神经监测技术在甲状腺手术中帮助识别喉上神经外支的临床意义

甲状腺癌手术中喉返神经损伤引起甲状腺外科医师的广泛关注,但喉上神经外支的保护常不为重视。喉上神经损伤导致声音嘶哑、发声疲劳、声频范围降低等症状。然而在甲状腺癌手术中肉眼识别喉上神经外支存在诸多困难,多数外科医师在常规暴露和保护神经方面显得力不从心。术中神经监测具有识别喉上神经外支以及评价其功能完整性的潜能。甲状腺癌手术中常规显露喉上神经外支,辅助术中神经监测,可显著降低术后神经损伤发生率,提高手术安全性。      

吸脂术对肋间臂神经保护的意义

 目的　探讨吸脂术在乳腺癌改良根治术中对保护肋间臂神经的意义。方法　2006年6月至2008年12月深圳市第六人民医院普外科诊治Ⅰ～ⅢA期乳腺癌患者共54例，随机分为吸脂组（脂肪切除术33例）和传统组（腋窝淋巴结切除术21例），对比分析两组手术时间、术中出血、肋间臂神经保护及术后随访等情况。结果　吸脂组手术时间长于传统组（P＜0.05），两组术中出血量比较差异无统计学意义（P＞0.05），吸脂组可明显提高肋间臂神经保护成功率（P＜0.05）、术后疼痛及麻木感降低（P＜0.05），两组随访6～24个月，均未见局部复发和远处转移。结论　在熟练掌握该技术的情况下，吸脂术可明显改善术中肋间臂神经损伤，可减少患者术后麻痹、疼痛。     

基于网络药理学和分子对接法预测甘草干姜汤抗乳腺癌的主要活性成分及作用机制

目的： 利用网络药理学和分子对接法研究甘草干姜汤（LDGD）可能的抗乳腺癌活性成分及作用机制。方法： 借助TCMSP数据库筛选出复方甘草干姜汤的主要有效成分;利用Genecards人类基因数据库获取乳腺癌的疾病靶点;利用STRING数据库和Cytoscape 3.8.0软件进行蛋白相互作用网络的构建和分析;并通过DAVID数据库进行GO富集分析和KEGG通路富集分析;基于文献检索甘草干姜汤的药效成分并将筛选出的关键靶点进行分子对接。结果： LDGD抗乳腺癌潜在的关键靶点有AKT1、MYC、TP53、EGF等;GO功能富集分析显示与作用靶点有关的生物学过程主要有凋亡信号通路、氧化应激等;细胞成分主要有细胞周期蛋白依赖性激酶、转录因子复合体、丝氨酸/苏氨酸蛋白激酶复合物、膜区、膜微区等;分子功能主要有蛋白质丝氨酸/苏氨酸激酶活性、转录因子活性核受体活性、泛素蛋白连接酶结合等;KEGG通路分析结果显示主要涉及p53、TNF、IL-17、EGFR、细胞凋亡等信号通路;以AKT1为靶点,8种药效成分中与其结合的小分子有甘草苷、6-姜酚、6-姜烯酚、甘草素、异甘草素、异甘草苷,其中甘草苷、6-姜酚、6-姜烯酚、甘草素的打分值大于原配体打分值的80%。结论： 甘草苷、6-姜酚、6-姜烯酚、甘草素可能是甘草干姜汤中抗乳腺癌主要活性成分之一,抗肿瘤作用机制可能涉及细胞增殖、凋亡、氧化应激、炎症等相关通路。     

诱导生物合成金属硫蛋白对顺铂引起血液毒性的保护作用

顺铂对小鼠血液系统具有毒副作用，而组织中金属硫蛋白（ＭＴ）的产生能减轻其毒性反应。观察多种含微量元素药物（甘草锌、富硒麦芽和亚硒酸钠）对诱导组织产生ＭＴ的作用和对顺铂引起血液系统毒性的保护作用。结果表明：富硒麦芽的ＭＴ诱导作用最强（Ｐ＜０．０１），其次为亚硒酸钠和甘草锌。甘草锌、富硒麦芽和亚硒酸钠对顺铂引起的小鼠血液系统毒性具有明显的保护作用。由此提示了在应用含锌、硒等微量元素药物降低顺铂引起重金属中毒时，诱导生物合成ＭＴ增加，可能是其中的重要机理之一。     

神经保护在乳腺癌改良根治术中的临床意义

目的:探讨乳腺癌改良根治术行保留胸前神经及肋间臂神经的方法及临床价值。方法:选择乳腺癌患者112例,保留胸前神经及肋间臂神经72例（试验组）,切断肋间臂神经及胸前神经40例（对照组）,观察随访两组术后情况。结果:神经保护能够明显减少术后6个月胸肌萎缩、运动及感觉障碍的发生率。两组胸肌萎缩发生率比较,有统计学差异(P<0.01);两组运动及感觉障碍有统计学差异（P<0.01）。结论:保留胸前神经及肋间臂神经的乳腺癌改良根治术能有效防止胸大肌萎缩和患侧腋窝上肢感觉及运动障碍发生。     

乳腺癌Auchincloss术中保护肋间臂神经、胸肌神经的意义

目的：探讨在乳腺癌Auchincloss术中保护肋间臂神经、胸肌神经的临床意义。方法：对2008年9月-2010年10月间的38例乳腺癌患者,在Auchincloss术中行腋淋巴结清扫时,注意游离并保护肋间臂神经、胸肌神经,随访观察术前、术后患者胸大肌功能、胸大肌外缘厚度、上臂内侧及腋部皮肤感觉功能的变化;腋窝淋巴结清扫的数量,对术中保护肋间臂神经、胸肌神经的价值进行评估。结果：38例患者中患侧上臂内侧及腋部皮肤感觉正常32例,感觉异常仅2例,占5.6%,4例腋窝淋巴结明显肿大与之黏连,放弃保留肋间臂神经;38例患者均成功保留胸肌神经,经术后随访观察,胸大肌功能均为5级,术后6个月复查B超,胸大肌外缘厚度与术前比较无明显差异。结论：在乳腺癌Auchincloss术中注意保护肋间臂神经、胸肌神经可有效避免术后上臂内侧皮肤感觉障碍及胸大肌萎缩,能明显改善患者术后生存质量,对手术疗效并无影响。     

乳腺癌Auchincloss术中保护肋间臂神经、胸肌神经的意义

目的:探讨在乳腺癌Auchincloss术中保护肋间臂神经、胸肌神经的临床意义.方法:对2008年9月-2010年10月间的38例乳腺癌患者,在Auchincloss术中行腋淋巴结清扫时,注意游离并保护肋间臂神经、胸肌神经,随访观察术前、术后患者胸大肌功能、胸大肌外缘厚度、上臂内侧及腋部皮肤感觉功能的变化;腋窝淋巴结清扫的数量,对术中保护肋间臂神经、胸肌神经的价值进行评估.结果:38例患者中患侧上臂内侧及腋部皮肤感觉正常32例,感觉异常仅2例,占5.6%,4例腋窝淋巴结明显肿大与之黏连,放弃保留肋间臂神经;38例患者均成功保留胸肌神经,经术后随访观察,胸大肌功能均为5级,术后6个月复查B超,胸大肌外缘厚度与术前比较无明显差异.结论:在乳腺癌Auchincloss术中注意保护肋间臂神经、胸肌神经可有效避免术后上臂内侧皮肤感觉障碍及胸大肌萎缩,能明显改善患者术后生存质量,对手术疗效并无影响.     

神经导航在经单鼻孔蝶窦入路手术中的应用——附78例报告

背景与目的:垂体瘤是颅内最常见的肿瘤之一,以手术治疗为主。保护肿瘤周围正常结构,顺利暴露肿瘤是经鼻蝶窦入路垂体瘤切除术的关键。本文旨在探讨不同影像资料下神经导航在经单鼻孔蝶窦入路手术中的应用。方法:选择临床诊断为垂体瘤患者78例。其中47例患者均应用神经CT导航辅助手术切除垂体瘤,31例患者均在神经MRI导航下,辅助手术切除垂体瘤,对两种手术方法的术中准确性及手术效果进行评价。结果:应用神经导航辅助手术切除78例垂体瘤的过程中均准确找到病灶,导航误差范围1.4～2.9mm。结论:经鼻蝶窦入路垂体瘤切除术中应用神经导航可靠、准确性高,同时可提高手术的疗效。     

甘草素通过影响鼻咽癌CNE-2 细胞的自噬增强其放疗敏感性

[摘要] 目的：探讨甘草素（licorice）对鼻咽癌CNE-2 细胞放疗增敏的影响及其作用机制。方法：体外培养构建放射抵抗性的鼻咽癌细胞株CNE-2-RR。MTT细胞实验检测不同浓度的甘草素对鼻咽癌细胞增殖活性的影响,透射电镜检测甘草素处理鼻咽癌细胞后自噬体的变化情况,Western blotting 检测甘草素对鼻咽癌细胞自噬蛋白水平的影响,彗星实验检测不同组鼻咽癌细胞DNA损伤修复情况,流式细胞术检测鼻咽癌细胞株凋亡率的变化。结果：成功构建放射抵抗细胞株CNE-2-RR,20 mmol/L甘草素对鼻咽癌细胞的最高抑制率为（58.86±5.02）%。甘草素处理鼻咽癌CNE-2-RR细胞后胞内自噬体数量增加,线粒体和细胞核形态异常;细胞中自噬体蛋白LC3-II 水平升高、LC3-I 水平降低（P<0.05）;甘草素作用CNE-2-RR细胞后彗星尾距长度大于对照组,表明对DNA损伤修复能力明显降低。甘草素作用导致CNE-2-RR细胞的凋亡率明显增加（P<0.05）。结论：甘草素通过影响鼻咽癌CNE-2-RR细胞的自噬行为及DNA修复能力增强其对放疗的敏感性。     

4-Methyl-3-nitro-benzoic acid, a migration inhibitor, prevents breast cancer metastasis in SCID mice

The aim of this study was to determine the anticancer effects of seven licorice compounds in MKN-28, AGS, and MKN-45 gastric cancer cells and human gastric epithelium immortalized cells. We also explored the mechanism of action of licochalcone A (LCA), the most cytotoxic licorice compound, by analyzing its influence on cell cycle progression and apoptosis. The results indicated that LCA was the most cytotoxic licorice compound of those tested, and it inhibited gastric cancer cells growth in a dose-dependent manner, with an IC50 value of approximately 40μM. LCA affected gastric cancer cell viability by blocking cell cycle progression at the G2/M transition and inducing apoptosis. LCA treatment increased the expression of Rb and decreased the expression of cyclin A, cyclin B and MDM2 in MKN-28, AGS and MKN-45 cell lines. In addition, LCA-induced apoptosis by its effects on the expression of PARP, caspase-3, Bcl-2 and Bax. These data provide evidence that LCA has the potential to be used in the treatment of gastric cancer.     

Effect of licorice compounds licochalcone A, glabridin and glycyrrhizic acid on growth and virulence properties of Candida albicans

Candida albicans is the predominant causal agent of candidiasis. Its ability to form hyphae and biofilm has been suggested to be key virulence factors. In this study, we investigated the effect of major licorice compounds licochalcone A, glabridin and glycyrrhizic acid on growth, biofilm formation and yeast-hyphal transition of C. albicans. The synergistic effect of licorice compounds with the antifungal drug nystatin was also evaluated. Minimal inhibitory concentrations (MICs) for C. albicans were determined using a microplate dilution assay. The synergistic effect with nystatin was determined similarly. The effect of licorice compounds on biofilm formation was evaluated using a microplate assay and crystal violet staining. The effect of licorice compounds on yeast-hyphal transition was determined by microscopic observation. The toxicity of licorice compounds towards oral epithelial cells was evaluated with an MTT assay. Glabridin and licochalcone A showed antifungal activity on C. albicans while glycyrrhizic acid had no effect. Complete growth inhibition occurred with sub-inhibitory concentrations of nystatin with either glabridin or licochalcone A. Biofilm formation was inhibited by 35-60% in the presence of licochalcone A (0.2 μg ml(-1)). A strong inhibitory effect (>80%) on hyphal formation was observed with licochalcone A or glabridin (100 μg ml(-1)). Glabridin and licochalcone A at high concentrations showed toxicity towards oral epithelial cells. In summary, glabridin and licochalcone A are potent antifungal agents and may act in synergy with nystatin to inhibit growth of C. albicans. Licochalcone A has a significant effect on biofilm formation, while both licochalcone A and glabridin prevented yeast-hyphal transition in C. albicans. These results suggest a therapeutic potential of licochalcone A and glabridin for C. albicans oral infections.     

Glabridin inhibits migration, invasion, and angiogenesis of human non-small cell lung cancer A549 cells by inhibiting the FAK/rho signaling pathway

This study reports the antimigration, anti-invasive effect of glabridin, a flavonoid obtained from licorice, in human non-small cell lung cancer A549 cells. Glabridin exhibited effective inhibition of cell metastasis by decreasing cancer cell migration and invasion of A549 cells. In addition, glabridin also decreased A549-mediated angiogenesis. Further investigation revealed that glabridin's inhibition of cancer angiogenesis was also evident in a nude mice model. Blockade of A549 cells migration was associated with an increase of ανβ3 integrin proteosome degradation. Glabridin also decreased the active forms of FAK and Src, and enhanced levels of inactivated phosphorylated Src (Tyr 527), decreasing the interaction of FAK and Src. Inhibition of the FAK/Src complex by glabridin also blocked Akt activation, resulting in reduced activation of RhoA and myosin light chain phosphorylation. This study demonstrates that glabridin may be a novel anticancer agent for the treatment of lung cancer in 3 different ways: inhibition of migration, invasion, and angiogenesis.     

Induction of apoptosis in human leukemia cells by MCS-C2 via caspase-dependent Bid cleavage and cytochrome c release

Much of the interest on the chemopreventive properties of licorice has been focused on the plant genius Glycyrrhiza glabra. In this study the ethanol extract of Chinese licorice root, Glycyrrhiza uralensis (G. uralensis) was investigated for its estrogenic effect and the ability to inhibit cell proliferation in the MCF-7 human breast cancer cell line. The extract of the root of G. uralensis was fractionated in EtOH:H₂O (80:20) (80% ethanol). The extract exhibited estrogenic effects similar to 17β- estradiol (E2) and induced apoptosis at the same dose level (100 μg/ml) in MCF-7 breast cancer cells, results were associated with up-regulation of tumor suppressor gene p53 and pro-apoptotic protein Bax. G. uralensis extract caused the up-regulation of p21^waf1/cip1 and down-regulation of cdk 2 and cyclin E and most significantly, induced G1 cell cycle arrest. This is the first study to show that the ethanolic extract of the root of G. uralensis has an estrogen-like activity and anti-cancer effects against MCF-7 human breast cancer cells. Whilst the use of phytoestrogens to protect against hormone-dependent cancers or as a ‘natural’ alternative to hormone replacement therapy remains controversial, the data in this paper support the suggestion that extracts of root of the Chinese licorice G. uralensis might be of importance in this debate.     

18α-glycyrrhetinic acid targets prostate cancer cells by down-regulating inflammation-related genes

Glycyrrhetinic acid is an active triterpenoid metabolite of glycyrrhizin abundantly present in licorice roots. Glycyrrhetinic acid exists as α and β stereo-isomeric forms. Both stereo-isomeric forms are known to have anti-inflammatory and anticancer activity. However, the effects and anticancer mechanism of α glycyrrhetinic acid in prostate cancer cells has not yet been evaluated. Therefore, we investigated the growth inhibition, induction of apoptosis and the anticancer mechanisms of 18α-glycyrrhetinic acid (AGA), on the androgen-independent metastatic prostate cancer cell line DU-145. Our results showed that AGA inhibited proliferation and growth of these cells by inducing apoptosis as determined by Annexin V and flow cytometry analyses. Our studies also showed that HUVEC tube formation was drastically reduced when cultured in conditioned medium of AGA-treated DU-145 cells. In addition, AGA treatment prevented the invasion of DU-145 prostate cancer cells on matrigel coated transwells via down-regulation of NF-κB (p65), VEGF and MMP-9 expression. Furthermore, AGA treatment also down-regulated the expression of pro-inflammatory cytokine/growth factor genes HMGB1, IL-6 and IL-8 in DU-145 cells. Interestingly, AGA simultaneously upregulated the expression of non-steroidal anti-inflammatory gene-1 (NAG-1) in DU-145 cells suggesting its anti-inflammatory activity on prostate cancer cells. Taken together, the results of this study suggest that AGA may be a promising anticancer agent that merits further investigation for the chemoprevention and treatment of prostate cancer.     

Antigen levels of the urokinase-type plasminogen activator and its gene polymorphisms in colorectal cancer

Isoliquiritigenin is a chalcone isolated from licorice and shallots. The ability of isoliquiritigenin to suppress metastasis was examined in a pulmonary metastasis model of mouse renal cell carcinoma. Isoliquiritigenin significantly reduced pulmonary metastasis, without any weight loss or leukocytopenia. Isoliquiritigenin suppressed in vitro proliferation of carcinoma cells, potentiated nitric oxide production by lipopolysaccharide-stimulated macrophages, and facilitated cytotoxicity of splenic lymphocytes in vitro. These findings suggest activation of macrophages, activation of cytotoxicity of lymphocytes, and direct cytotoxicity as possible mechanisms of metastasis suppression by isoliquiritigenin. In addition, isoliquiritigenin prevented severe leukocytopenia caused by administration of 5-fluorouracil.     

11种天然药材及植物的抗突变与致突变同步试验报告

本文报告用ＳＯＳ噬菌体诱导试验的抗突变与致突变同步试验法，对１１种天然中药及植物甘草、枸杞、半枝莲、柴胡、丹参、黄芪、决明子、仙人球、仙人指、胡萝卜及青萝卜的抗突变及致突变性进行了筛检。经过加和不加大鼠肝微粒体酶代谢活化系统（Ｓ９）的２种试验及重复验证，结果１１种中药及植物均未发现致突变作用，甘草、枸杞、丹参、黄芪、胡萝卜、青萝卜及仙人指可抑制抗肿瘤药物丝裂霉素Ｃ（ＭＭＣ）的致突变性。     

Inhibition of P-glycoprotein and multidrug resistance protein 1 by dietary phytochemicals

PURPOSE: For the development of a safe and effective dual inhibitor of anticancer drug efflux transporters P-glycoprotein and multidrug resistance protein 1 (MRP1) to conquer multidrug resistance, we investigated the effects of dietary phytochemicals on the functions of P-glycoprotein and MRP1. METHODS: The effects of dietary phytochemicals on the functions of P-glycoprotein and MRP1 were investigated using P-glycoprotein-overexpressing human carcinoma KB-C2 cells and human MRP1 gene-transfected KB/MRP cells. The effects of natural compounds found in dietary supplements, herbs, and foods such as sesame, ginkgo, soybean, and licorice were evaluated. RESULTS: The accumulation of daunorubicin, a fluorescent substrate of P-glycoprotein, increased in the presence of sesamin, ginkgolic acid, matairesinol, glycyrrhetinic acid, glabridin, and phyllodulcin in KB-C2 cells. Glycyrrhetinic acid and matairesinol also increased the accumulation of calcein, a fluorescent substrate of MRP1, in KB/MRP cells. KB-C2 and KB/MRP cells were sensitized to anticancer drugs by glycyrrhetinic acid, showing that glycyrrhetinic acid reverses multidrug resistance. The verapamil-stimulated P-glycoprotein ATPase activity was inhibited by glycyrrhetinic acid. Glycyrrhetinic acid stimulated the ATPase activity of MRP1. CONCLUSION: These results suggest that dietary phytochemicals, such as glycyrrhetinic acid found in licorice, have dual inhibitory effects on P-glycoprotein and MRP1 and might become useful to enhance the efficacy of cancer chemotherapy.     

Inhibition of protein kinase C by the 12-O-tetradecanoylphorbol-13-acetate antagonist glycyrrhetic acid

Glycyrrhetic acid is an anti-inflammatory agent isolated from licorice root that inhibits 12-O-tetradecanoylphorbol-13-acetate (TPA)-mediated tumor promotion in mouse skin. Although it has been established that glycyrrhetic acid inhibits a number of events induced by the phorbol ester tumor promoter TPA in cultured cells, its mechanisms of action has remained obscure. In this report, we demonstrate that glycyrrhetic acid inhibits the Ca2+-and phospholipid-dependent phosphotransferase activity of protein kinase C (PKC), the phorbol ester tumor promoter receptor. Therefore, inhibition of PKC may play a role in the anti-promoting activity of glycyrrhetic acid.