Glabridin induces apoptosis and cell cycle arrest in oral cancer cells through the JNK1/2 signaling pathway

Glabridin, a flavonoid extracted from licorice (Glycyrrhiza glabra), possesses various biological properties, including anticancer activities. However, the effect of glabridin on oral cancer cell apoptosis and the underlying molecular mechanisms has not been elucidated. In this study, we demonstrated that glabridin treatment significantly inhibits cell proliferation in human oral cancer SCC‐9 and SAS cell lines. Flow cytometric assays demonstrated that glabridin induced several features of apoptosis, such as sub‐G1 phase cell increase and phosphatidylserine externalization. Furthermore, glabridin induced apoptosis dose‐dependently in SCC‐9 cells through caspase‐3, −8, and −9 activation and poly (ADP‐ribose) polymerase cleavage. Moreover, glabridin increased the phosphorylation of the extracellular signal–regulated kinase, p38, and c‐Jun N‐terminal kinase (JNK) pathways in a dose‐dependent manner. Moreover, the inhibition of the JNK1/2 inhibitor significantly reversed the glabridin‐induced activation of the caspase pathway. In conclusion, our findings suggest that glabridin induces oral cancer cell apoptosis through the JNK1/2 pathway and is a potential therapeutic agent for oral cancer.     

光甘草定通过调节ERK/IRS-1和PI3K/Akt信号通路改善HepG2细胞的胰岛素抵抗

目的 探索光甘草定改善肝细胞胰岛素抵抗（insulin resistance,IR）的作用和机制。方法 通过高胰岛素诱导人肝癌HepG2细胞建立IR模型,采用葡萄糖氧化酶法检测细胞的葡萄糖消耗量及生成;荧光标记法检测葡萄糖摄取量;蒽酮法检测糖原含量;ELISA检测葡萄糖代谢关键酶的活性;Western blotting检测磷脂酰肌醇3-激酶/蛋白激酶B（phosphatidylinositol3-kinase/protein kinase B,PI3K/Akt）、细胞外调节蛋白激酶/胰岛素受体底物-1（extracellular regulated protein kinase/insulinreceptor substrate-1,ERK/IRS-1）信号通路相关蛋白以及葡萄糖转运蛋白4（glucose transporter 4,GLUT4）的表达。采用分子对接技术研究光甘草定和ERK分子间的相互作用。结果 光甘草定显著增加IR-HepG2细胞的葡萄糖消耗和摄取（P＜0.05）;通过显著提高糖原合成酶（glycogen synthase,GS）、葡萄糖激酶（glucokinase,GCK）和丙酮酸激酶（pyruvate kinase,PK）活性（P＜0.05、0.01）,促进IR-HepG2细胞的糖原合成和糖酵解;通过显著减弱磷酸烯醇丙酮酸羧激酶（phosphoenolpyruvate carboxykinase,PEPCK）和葡萄糖-6-磷酸酶（glucose-6-phosphatase,G6Pase）的活性（P＜0.05）,抑制IR-HepG2细胞的糖异生。IR-HepG2细胞经光甘草定处理后,Akt、糖原合成酶激酶-3β（glycogen synthase kinase-3β,GSK-3β）和叉头框蛋白O1（forkhead boxing protein O1,FOXO1）的磷酸化水平得到显著恢复（P＜0.01）,而这种作用被PI3K的抑制剂LY294002所逆转（P＜0.01）。同时,光甘草定显著促进GLUT4向质膜的易位（P＜0.01）。光甘草定显著降低IRHepG2细胞的ERK和IRS的磷酸化水平（P＜0.01）,还可作为ERK的I1/2型抑制剂。结论 光甘草定通过抑制ERK/IRS-1通路,激活PI3K/Akt信号通路,修复IR-HepG2细胞的糖代谢紊乱,缓解IR症状。     

光甘草定醇质体与立方液晶纳米粒皮肤滞留量比较及抗豚鼠皮肤光老化作用观察

目的 制备光甘草定醇质体与立方液晶纳米粒,通过测定2种纳米制剂在离体豚鼠皮肤中的滞留量优选出适合光老化经皮给药的制剂。建立光老化模型,通过观察优选制剂对豚鼠光老化模型的治疗效果来评价药物的疗效。方法 采用注入法制备光甘草定醇质体,高压均质法制备光甘草定立方液晶纳米粒,利用Franz扩散装置考察光甘草定醇质体、立方液晶纳米粒凝胶离体鼠皮中的滞留量,优选出光甘草定经皮给药治疗光老化的纳米制剂。紫外线照射背部剃毛豚鼠建立皮肤光老化模型。雌性豚鼠随机分为模型组、基质（给予空白凝胶0.5 g/只）组、维甲酸（阳性对照,0.5 g/只）组和甘草定立方液晶纳米粒凝胶高、低剂量（0.50、0.25 g/只）组,治疗2周后,通过肉眼观察、HE染色、Masson染色等评价其治疗效果,用水份测定仪观察其对豚鼠皮肤含水量的影响。结果 2种纳米制剂凝胶的鼠皮滞留量最高者为光甘草定立方液晶纳米粒凝胶。豚鼠皮肤光老化模型建立成功,HE染色、Masson染色等结果表明光甘草定立方液晶纳米粒对光老化有明显的治疗效果,使光老化皮肤的含水量显著升高（P<0.05）。结论 光甘草定立方液晶纳米粒在离体鼠皮中的滞留较高,对豚鼠皮肤光老化模型治疗效果显著,为光甘草定的临床应用提供了新的方法与思路。     

Review of pharmacological effects of Glycyrrhiza sp. and its bioactive compounds

The roots and rhizomes of licorice (Glycyrrhiza) species have long been used worldwide as a herbal medicine and natural sweetener. Licorice root is a traditional medicine used mainly for the treatment of peptic ulcer, hepatitis C, and pulmonary and skin diseases, although clinical and experimental studies suggest that it has several other useful pharmacological properties such as antiinflammatory, antiviral, antimicrobial, antioxidative, anticancer activities, immunomodulatory, hepatoprotective and cardioprotective effects. A large number of components have been isolated from licorice, including triterpene saponins, flavonoids, isoflavonoids and chalcones, with glycyrrhizic acid normally being considered to be the main biologically active component. This review summarizes the phytochemical, pharmacological and pharmacokinetics data, together with the clinical and adverse effects of licorice and its bioactive components.     

Inhibition of serotonin re-uptake by licorice constituents

Ovarian steroid hormones, estrogen and progestin, affect the function of the serotonin neural system by inhibiting serotonin re-uptake through allosteric interaction with the serotonin transporter (SERT) in a nongenomic mechanism. Blocking or reducing serotonin re-uptake at the synapse alleviates depression. The aim of this study was to test the effect of compounds of the isoflavan and isoflavene groups, subclasses of the flavonoids family, on serotonin re-uptake and to compare the results with the effect of other known phytoestrogens like genistein and daidzein to relate the activity of these compounds to their structure. The effect of these compounds on the re-uptake of radioactive serotonin was assayed in HEK-293 cells stably expressed the recombinant human serotonin transporter (hSERT). The results demonstrated that the isoflavans glabridin and 4'-O-methylglabridin (4'-OMeG) and the isoflavene glabrene inhibited serotonin re-uptake by 60, 53 and 47%, respectively, at 50 microM, whereas resorcinol, the isoflavan 2'-O-methylglabridin (2'-OMeG), and the isoflavones genistein and daidzein were inactive. The inhibition of serotonin re-uptake is dose dependant with glabridin and estradiol. These results emphasize the importance of the lipophilic part of the isoflavans, as well as the hydroxyl at position 2' on ring B. In conclusion, this study showed that several isoflavans are unique phytoestrogens, which like estradiol, affects the serotonergic system and inhibits serotonin re-uptake and, thus, potentially may be beneficial for mild to moderate depression in pre- and postmenopausal women.     

促渗剂对光甘草定体外经皮渗透效果的影响

目的:考察促渗剂对光甘草定体外经皮渗透性能的影响,为该化合物经皮给药系统的开发提供参考。方法:采用HPLC测定光甘草定含量,流动相乙腈-水-冰乙酸(55∶44∶1),检测波长282nm。选取小鼠背部皮肤,通过Franz垂直扩散池考察光甘草定的透皮性能,考察氮酮、冰片、丙二醇对光甘草定乳剂透皮吸收的影响。结果:光甘草定的透皮吸收率较低,促渗剂能有效使其透过皮肤。不同促渗剂对光甘草定促透作用顺序为5%氮酮组>5%丙二醇组>5%冰片组>单体组,稳态渗透速率分别为4.938,3.041,2.950,2.583μg·cm-2·h-1,滞后时间依次为0.056,0.180,0.059,0.167h。结论:不同促渗剂均能促进光甘草定的经皮渗透效果,以氮酮为最佳。     

光甘草定对宫颈癌Hela细胞增殖、侵袭和凋亡的影响及其机制

目的:探讨光甘草定对宫颈癌Hela细胞增殖、侵袭和凋亡的影响及其机制。方法:以0、1、2.5、5、10和20 μmol/L光甘草定作用于体外培养的Hela细胞24 h后，采用细胞计数（CCK-8)法检测细胞存活率并计算出光甘草定的半抑制浓度。将体外培养的Hela细胞分别以0、2.5和5 μmol/L光甘草定处理后，采用Transwell小室检测细胞侵袭变化，流式细胞仪检测细胞凋亡和周期变化，免疫印迹法检测细胞中Wnt/β-连环蛋白（β-catenin）信号通路相关蛋白Wnt1、β-catenin、细胞周期蛋白D1（CyclinD1）、基质金属蛋白酶-2（MMP-2）和生存素（Survivin）的表达情况。结果:与对照（0 μmol/L）组相比，1、2.5、5、10和20 μmol/L 光甘草定处理组细胞存活率均明显降低（P<0.05），且呈浓度依赖性；同时，光甘草定对Hela细胞的半抑制浓度为4.56 μmol/L。与对照（0 μmol/L）组相比，2.5和5 μmol/L光甘草定处理组中侵袭细胞数和细胞在S期与G2/M期的百分比以及细胞中Wnt1、β-catenin、CyclinD1、MMP-2、Survivin蛋白的表达水平均明显减少，而细胞凋亡率和细胞在G0/G1期百分比明显升高（P<0.05），且5 μmol/L光甘草定处理组细胞的变化更为显著。结论:光甘草定可抑制Hela细胞增殖、侵袭并促进其凋亡，其作用机制可能与抑制Wnt/β-catenin信号通路的活化有关。     

Retracted: Glabridin attenuates the migratory and invasive capacity of breast cancer cells by activating microRNA‐200c

Current treatments for breast cancer, a common malignancy in human females, are less than satisfactory because of high rates of metastasis. Glabridin (GLA), which acts through the FAK/ROS signaling pathway, has been used as an antioxidant and anti‐metastatic agent. However, little is known regarding the effect of microRNA (miRNA) on GLA's anti‐metastatic activity. The miRNA‐200 family, which is frequently expressed at low levels in triple negative breast cancers, inhibits metastasis by blocking the epithelial–mesenchymal transition. Here, we found that GLA attenuated the migratory and invasive capacity of breast cancer cells by activating miR‐200c. GLA induced the mesenchymal–epithelial transition in vitro and in vivo, as determined by increased expression of the epithelial marker, E‐cadherin, and decreased expression of the mesenchymal marker, vimentin. Overexpression of miR‐200c enhanced the expression of E‐cadherin and decreased the expression of vimentin. Furthermore, in MDA‐MB‐231 and BT‐549 breast cancer cells exposed to GLA, knockdown of miR‐200c blocked the GLA‐induced mesenchymal–epithelial transition and alleviated the GLA‐induced inhibition of migration and invasion. Thus, elevation of miR‐200c by GLA has considerable therapeutic potential for anti‐metastatic therapy for breast cancer patients.     

甘草及其黄酮类化合物的神经保护作用

甘草及其黄酮类化合物对神经元有保护作用,能对抗缺血再灌注引起的脑损伤,也有改善学习记忆和抗抑郁作用。综述甘草粗提物及其异甘草素、甘草素、甘草查耳酮、光甘草定等黄酮类化合物的神经保护作用的研究进展文献,认为甘草及其黄酮类化合物的神经保护作用是其抗炎、抗氧化、抗细胞凋亡及抗胆碱酯酶、单胺氧化酶、环磷酸腺苷磷酸二酯酶等基本药理作用综合的结果。     

RP-HPLC测定不同产地光果甘草废渣中光甘草定含量

目的利用反相高效液相色谱法测定由不同产地光果甘草制备的甘草废渣中光甘草定的含量。方法对4种产地的光果甘草用同一种水提法制备甘草废渣;再用不同提取法制备6种总黄酮粗提物;采用SymmetryC18反相色谱柱,流动相为乙腈-水-冰醋酸（55∶44∶1）,检测波长为282nm,流速为1.0mL·min^-1,柱温为30℃,测定各样品中光甘草定的含量。结果光甘草定在0.09～0.45mg·mL^-1内呈良好线性关系（r=0.9997）,平均回收率100.2%,RSD为0.89%。6种总黄酮粗提物中光甘草定含量分别为1.31%,1.16%,1.59%,2.93%,0.98%,0.85%;4种甘草废渣中,光甘草定含量分别为0.225%,0.290%,0.211%,0.218%。结论反相高效液相色谱法简便、快速、重复性好,适用于光果甘草中光甘草定的含量测定。4种光果甘草废渣中,新疆和静的光果甘草废渣所含光甘草定含量明显高于其他产地的光果甘草废渣。此外,利用甲醇回流提取法制备光甘草定时,提取率和总黄酮出膏率都高于乙醇回流提取法和甲醇超声提取法。