Nanomedicine-boosting icaritin-based immunotherapy of advanced hepatocellular carcinoma

Traditional treatments for advanced hepatocellular carcinoma(HCC),such as surgical resection,transplantation,radiofrequency ablation,and chemotherapy are unsatisfactory,and therefore the exploration of powerful therapeutic strategies is urgently needed.Immunotherapy has emerged as a promising strategy for advanced HCC treatment due to its minimal side effects and long-lasting therapeutic memory effects.Recent studies have demonstrated that icaritin could serve as an immunomodulator for effective immunotherapy of advanced HCC.Encouragingly,in 2022,icaritin soft capsules were approved by the National Medical Products Administration(NMPA)of China for the immunotherapy of advanced HCC.However,the therapeutic efficacy of icaritin in clinical practice is impaired by its poor bioavailability and unfavorable in vivo delivery efficiency.Recently,functionalized drug delivery systems including stimuli-responsive nanocarriers,cell membrane-coated nanocarriers,and living cell-nanocarrier systems have been designed to overcome the shortcomings of drugs,including the low bioavailability and limited delivery efficiency as well as side effects.Taken together,the development of icaritin-based nanomedicines is expected to further improve the immunotherapy of advanced HCC.Herein,we compared the different preparation methods for icaritin,interpreted the HCC immune microenvironment and the mechanisms underlying icaritin for treatment of advanced HCC,and discussed both the design of icaritin-based nanomedicines with high icaritin loading and the latest progress in icaritinbased nanomedicines for advanced HCC immunotherapy.Finally,the prospects to promote further clinical translation of icaritin-based nanomedicines for the immunotherapy of advanced HCC were proposed.

Anticancer Effect of Icaritin on Human Lung Cancer Cells through Inducing S Phase Cell Cycle Arrest and Apoptosis

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.

Icaritin requires phosphatidylinositol 3kinase/Akt signaling to counteract skeletal muscle atrophy following mechanical unloading

OBJECTIVE Skeletal muscle undergoes rapid and profound atrophy in response to decreased mechanical loading,e.g.,limb immobilization and bed rest.Phosphatidylinositol 3 kinase(PI3K)/Akt signaling pathway is critical for regulating the balance between protein synthesis and degradation during disuse/inactivity-induced muscle atrophy.The present study aimed to investigate whether natural product Icaritin(ICT)required PI3K/Akt signaling to exert counteractive effect on skeletal muscle atrophy following mechanical unloading.METHODS Two oral dosages of ICT(80and 120mg·kg-1·d-1)were administrated daily to adult male rats with or without daily injection of PI3K/Akt signaling inhibitor wortmannin(15μg·kg-1·d-1)during 28-d hindlimb suspension(HS).Ex vivo muscle functional testing,histological and immunohistochemical analysis were performed to determine the changes of soleus muscle function,mean muscle fiber cross-sectional area(CSA)and fiber type distribution.Western blot and real-time PCR analysis were also performed to evaluate the protein or mRNA expression of the markers involved in PI3K/Akt signaling pathway.RESULTS After 28-d HS,soleus muscle underwent profound muscle atrophy(-52.7% muscle mass vs.pre-HS baseline).The high dose ICT treatment significantly attenuated the decreases in soleus muscle mass(+22.6% vs.HS),muscle fiber CSA(+52.8% vs.HS),as well as the muscle functional testing parameters during the unloading.Molecularly,the high dose ICT treatment significantly attenuated the decreases in protein synthesis markers at protein levels(phosphorylation of Akt and its downstream proteins)during the unloading,whereas the increases in protein degradation markers at mRNA(atrogin-1and MuRF-1)and protein(nuclear FOXO1 and FOXO3a)levels during the unloading were significantly attenuated by the high dose ICT treatment.The low dose ICT treatment moderately attenuated the above changes induced by the unloading.Mechanistically,Wortmannin could abolish the above effects of ICT.CONCLUSION ICT requires participation of PI3K/Akt signaling to counteract skeletal muscle atrophy following mechanical unloading in a dose-dependent manner.

Icaritin inhibits the progression of urothelial cancer by suppressing PADI2-mediated neutrophil infiltration and neutrophil extracellular trap formation

Tumor relapse and metastasis are the major causes of mortality associated with urothelial cancer.In the tumor microenvironment,negative regulatory molecules and various immune cell subtypes suppress antitumor immunity.The inflammatory microenvironment,associated with neutrophils and neutrophil extracellular traps(NETs),promotes tumor metastasis.However,no drugs are currently available to specifically inhibit neutrophils and NETs.In this study,we first demonstrated that icaritin(ICT),a Chinese herbal remedy that is a first-line treatment for advanced and incurable hepatocellular carcinoma,reduces NETs caused by suicidal NETosis and prevents neutrophil infiltration in the tumor microenvironment.Mechanistically,ICT binds to and inhibits the expression of PADI2 in neutrophils,thereby suppressing PADI2-mediated histone citrullination.Moreover,ICT inhibits ROS generation,suppresses the MAPK signaling pathway,and inhibits NET-induced tumor metastasis.Simultaneously,ICT inhibits tumoral PADI2-mediated histone citrullination,which consequently suppresses the transcription of neutrophil-recruiting genes such as GM-CSF and IL-6.The downregulation of IL-6 expression,in turn,forms a regulatory feedback loop through the JAK2/STAT3/IL-6 axis.Through a retrospective study of clinical samples,we found a correlation between neutrophils,NETs,UCa prognosis,and immune evasion.Combining ICT with immune checkpoint inhibitors may have synergistic effects.In summary,our study demonstrated that ICT could be a novel inhibitor of NETs and a novel UCa treatment.

Enhancing osteogenic bioactivities of coaxial electrospinning nanoscaffolds through incorporating iron oxide nanoparticles and icaritin for bone regeneration

Bone tissue engineering provides a promising strategy for the treatment of bone defects.Nonetheless,the clinical utilization of biomaterial-based scaffolds is constrained by their inadequate mechanical strength and absence of osteo-inductive properties.Here,we proposed to endow nano-scaffold(NS)constructed by coaxial electrospinning technique with enhanced osteogenic bioactivities and mechanical properties by incorporating biocompatible magnetic iron oxide nanoparticles(IONPs)and icaritin(ICA).Four types of nano-scaffolds(NS,ICA@NS,NS-IONPs and ICA@NS-IONPs)were prepared.The incorporation of ICA and IONPs minimally impact their surface morphological and chemical properties.IONPs enhanced the mechanical properties of NS scaffolds,including hardness,tensile strength,and elastic modulus.In vitro assessments demonstrated that ICA@NS-IONPs exhibited enhanced osteogenic bioactivities towards mouse calvarial pre-osteoblast cell line MC3T3-E1 as evidenced by detecting the alkaline phosphatase(ALP)activity level,expressions of osteogenesis-related genes and proteins as well as mineralized nodule formation.Mechanistic investigations revealed that MEK/ERK(MAP kinase-ERK kinase(MEK)/extracellularsignal-regulated kinase(ERK))signaling pathway could offer a plausible explanation for the osteogenic differentiation of MC3T3-E1 cells induced by ICA@NS-IONPs.Furthermore,the implantation of nano-scaffolds in rat skull defects exhibited a substantial improvement in in vivo bone regeneration.Therefore,IONPs and ICA incorporated coaxial electrospinning nano-scaffolds present a novel strategy for the optimization of scaffolds for bone tissue engineering.

Conversion therapy of a giant hepatocellular carcinoma with portal vein thrombus and inferior vena cava thrombus:A case report and review of literature

BACKGROUND The prognosis of hepatocellular carcinoma(HCC)combined with portal and hepatic vein cancerous thrombosis is poor,for unresectable patients the combination of targeted therapy and immune therapy was the first-line recommended treatment for advanced HCC,with a median survival time of only about 2.7-6 months.In this case report,we present the case of a patient with portal and hepatic vein cancerous thrombosis who achieved pathologic complete response after conversion therapy.CASE SUMMARY In our center,a patient with giant HCC combined with portal vein tumor thrombus and hepatic vein tumor thrombus was treated with transcatheter arterial chemoembolization(TACE),radiotherapy,targeted therapy and immunotherapy,and was continuously given icaritin soft capsules for oral regulation.After 7 months of conversion therapy,the patient's tumor shrank and the tumor thrombus subsided significantly.The pathology of surgical resection was in complete remission,and there was no progression in the postoperative follow-up for 7 months,which provided a basis for the future strategy of combined conversion therapy.CONCLUSION In this case,atezolizumab,bevacizumab,icaritin soft capsules combined with radiotherapy and TACE had a good effect.For patients with hepatocellular carcinoma combined with hepatic vein/inferior vena cava tumor thrombus,adopting a high-intensity,multimodal proactive strategy under the guidance of multidisciplinary team(MDT)is an important attempt to break through the current treatment dilemma.

淫羊藿活性成分抗卵巢癌作用机制研究进展

卵巢癌是女性生殖系统三大癌症之一,其病死率位居妇科恶性肿瘤之首。淫羊藿具有补肾阳、强筋骨、祛风湿的功效,临床常用于生殖系统疾病的治疗。本文通过综述国内外相关文献,发现淫羊藿主要活性成分淫羊藿苷、淫羊藿素和淫羊藿次苷Ⅱ,主要从促进细胞凋亡、影响细胞的侵袭和转移、阻滞细胞周期、诱导细胞自噬、促进细胞焦亡等5个方面发挥抗卵巢癌作用,可见淫羊藿可以通过多靶点、多途径治疗卵巢癌。

D-半乳糖对小鼠睾丸TM4支持细胞连接功能损伤及淫羊藿素的改善作用研究

目的研究D-半乳糖(D-galactose,D-gal)对小鼠睾丸TM4支持细胞连接功能损伤以及淫羊藿素(icaritin,ICT)的改善作用机制。方法首先将TM4细胞分为正常对照组、不同浓度D-gal处理组,Western blot检测TM4细胞连接功能相关蛋白(ZO-1、Occludin、β-catenin和Cx43)以及ERα/FAK信号通路相关蛋白(ERα、FAK和pY397-FAK)表达水平的变化;随后MTT筛选ICT药物浓度,并将TM4细胞分为正常对照组、D-gal处理组、D-gal处理+不同浓度ICT组,Western blot检测上述蛋白表达水平变化;分子对接研究ERα与ICT之间的相互作用,并预测两者之间的亲和力;最后将TM4细胞分为正常对照组、D-gal处理组、ERα抑制剂组、D-gal+ICT组、ERα抑制剂+ICT组,Western blot检测上述蛋白表达水平的变化。结果D-gal可浓度依赖性下调连接功能相关蛋白(ZO-1、Occludin、β-catenin和Cx43)以及ERα/FAK信号通路相关蛋白(ERα、FAK和pY397-FAK)表达水平;给予ICT药物后,上述蛋白表达水平均明显上调;ERα和ICT分子对接结果显示,ERα的Asp351氨基酸残基与ICT之间形成2个距离为3.4Å和2.4Å的氢键,且两者之间的对接结合能小于-7 kcal·mol^(-1);ERα抑制剂处理TM4细胞后,上述蛋白表达水平均明显下调,给予ICT后,上述蛋白表达水平未有明显变化。结论D-gal可导致TM4细胞连接功能损伤,而ICT可改善其损伤,机制可能与上调ERα/FAK信号通路相关。

淫羊藿素联合吗啡对骨癌痛小鼠的镇痛效应研究

目的:探究淫羊藿素对骨癌痛(bone cancer pain,BCP)小鼠的镇痛效应,以及淫羊藿素与吗啡联合用药的镇痛效应;确定半效剂量淫羊藿素与吗啡联合用药,在达到吗啡最大镇痛效应时所减少的吗啡用量。方法:使用雄性C57BL/6J小鼠及路易斯(Lewis)肺癌细胞建立BCP小鼠模型。通过测定机械刺激缩足反射阈值(mechanical withdrawal threshold,MWT),冷刺激缩足潜伏期(paw withdrawal latency,PWL)以及自发抬足次数和抬足保护时间以评估小鼠疼痛敏感度。结果:自造模后第7~14天起,BCP小鼠左侧(荷瘤侧)后爪MWT和PWL明显降低,自发抬足次数和抬足保护时间明显上升,并持续存在(P<0.05)。造模后第7~14天给予淫羊藿素,自第10~14天起淫羊藿素明显增加BCP小鼠左侧后爪的MWT和PWL,并减少自发抬足次数和抬足保护时间,停药后效应持续至造模后第18天(即停药后第4天)(P<0.05)。应用2倍EC50剂量淫羊藿素(200 mg/kg)与吗啡联合用药可明显延长停药后镇痛效应,停药后镇痛效应持续至造模后第21天(即停药后第7天)(P<0.05)。应用半效剂量淫羊藿素(100 mg/kg)与吗啡联合用药,达到吗啡最佳镇痛效应时最多可减少60%吗啡用量。结论:淫羊藿素对BCP小鼠有明显的镇痛效应,当与吗啡联合用药时,不仅明显延长了停药后的镇痛效应,还能够减少高达60%的吗啡用量。

淫羊藿苷对消化系统肿瘤作用及其机制研究进展

消化道肿瘤预后差且缺乏有效治疗药物,是导致人类死亡的主要原因之一。淫羊藿(黄檗科)作为补肾壮阳中药用药历史悠久,现代研究表明淫羊藿及其主要活性成分淫羊藿苷(Icaritin,ICA)具有多途径、多靶点的抗肿瘤特性,是有广泛应用前景的抗消化道肿瘤药物,其成药阿可拉定已在肝癌、宫颈癌患者中显示出良好疗效。通过梳理淫羊藿苷调节肿瘤细胞周期,抑制肿瘤细胞增殖和迁徙,调控肿瘤免疫等机制,综述近年来淫羊藿苷在消化系统肿瘤中的研究进展,以期加快淫羊藿苷药理作用分子机制的探索和发现,为临床新药研发提供参考依据。

基于网络药理学与体外实验探讨淫羊藿素抗非小细胞肺癌EGFR-TKIs耐药的分子机制

目的基于网络药理学与体外实验探讨淫羊藿素抗非小细胞肺癌(NSCLC)表皮生长因子受体-酪氨酸酶抑制剂(EGFR-TKIs)耐药的分子机制。方法利用PubChem和化合物靶点预测(Swiss Target Prediction)数据库下载淫羊藿素的简化分子线性输入规范(SMILES)号及作用靶点,通过人类基因(GeneCards)和在线人类孟德尔遗传(OMIM)数据库收集NSCLC耐药疾病靶点,将药物与疾病交集靶点导入STRING数据库分析蛋白质-蛋白质相互作用(PPI)情况,运用Cytoscape 3.9.1软件内置插件计算节点拓扑参数值并筛选核心靶点,采用生物信息学分析平台(DAVID)数据库进行京都基因与基因组百科全书(KEGG)和基因本体(GO)富集分析,构建“淫羊藿素-关键靶点-疾病-通路”图。使用Pymol和Autodock Tools 1.5.7软件进行分子对接。体外实验选用NSCLC耐药株PC9OR研究淫羊藿素对细胞增殖、集落形成、迁移、侵袭和凋亡能力的影响,并对富集得到的核心靶点及关键通路进行验证。结果共筛选到淫羊藿素治疗NSCLC耐药的潜在作用靶点1952个。通过PPI网络节点拓扑参数值筛选得到13个核心靶点,涉及蛋白激酶B(AKT1)、雌激素受体α(ESR1)、B淋巴细胞瘤2(BCL2)、表皮生长因子受体(EGFR)等基因。KEGG通路富集分析显示,癌症相关通路、磷脂酰肌醇-3-激酶-蛋白激酶B(PI3K-AKT)信号通路、EGFR-TKIs耐药通路等可能在淫羊藿素治疗NSCLC EGFR-TKIs耐药的过程中起关键作用。GO富集分析显示,细胞功能涉及信号传导、凋亡过程的负调控、DNA转录的正调控等。分子对接显示淫羊藿素与各核心靶点均具有较强的结合能力。细胞实验表明,淫羊藿素抑制耐药细胞的增殖、集落形成、迁移、侵袭及促进细胞凋亡,并下调ESR1、AKT1、EGFR等mRNA表达水平以及PI3K-AKT通路磷酸化磷脂酰肌醇-3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)的关键蛋白水平。结论淫羊藿素可能通过多靶点调控PI3K-AKT通路抑制EGFR-TKIs耐药细胞的增殖、集落形成、迁移、侵袭及促进细胞凋亡,从而发挥抗NSCLC EGFR-TKIs耐药的作用。

Reversal of multidrug resistance by icaritin in doxorubicin-resistant human osteosarcoma cells

Multidrug resistance(MDR) is one of the major obstacles in cancer chemotherapy. Our previous study has shown that icariin could reverse MDR in MG-63 doxorubicin-resistant(MG-63/DOX) cells. It is reported that icariin is usually metabolized to icariside II and icaritin. Herein, we investigated the effects of icariin, icariside Ⅱ, and icaritin(ICT) on reversing MDR in MG-63/DOX cells. Among these compounds, ICT exhibited strongest effect and showed no obvious cytotoxicity effect on both MG-63 and MG-63/DOX cells ranging from 1 to 10 μmol·L^(-1). Furthermore, ICT increased accumulation of rhodamine 123 and 6-carboxyfluorescein diacetate and enhanced DOX-induced apoptosis in MG-63/DOX cells in a dose-dependent manner. Further studies demonstrated that ICT decreased the m RNA and protein levels of multidrug resistance protein 1(MDR1) and multidrug resistance-associated protein 1(MRP1). We also verified that blockade of STAT3 phosphorylation was involved in the reversal effect of multidrug resistance in MG-63/DOX cells. Taken together, these results indicated that ICT may be a potential candidate in chemotherapy for osteosarcoma.

Statistically designed enzymatic hydrolysis of an icariin/b-cyclodextrin inclusion complex optimized for production of icaritin

This study focuses on the preparation and enzymic hydrolysis of an icariin/bcyclodextrin inclusion complex to efficiently generate icaritin.The physical characteristics of the inclusion complex were evaluated by differential scanning calorimetry(DSC).Enzymatic hydrolysis was optimized for the conversion of icariin/b-cyclodextrin complex to icaritin by Box–Behnken statistical design.The inclusion complex formulation increased the solubility of icariin approximately 17-fold,from 29.2 to 513.5 mg/mL at 60℃.The optimum conditions were predicted by Box–Behnken statistical design as follows:60℃,pH 7.0,the ratio of enzyme/substrate(1:1.1)and reaction time 7 h.Under the optimal conditions the conversion of icariin was 97.91%and the reaction time was decreased by 68%compared with that without b-CD inclusion.Product analysis by melting point,ESI-MS,UV,IR,1H NMR and 13C NMR confirmed the authenticity of icaritin with a purity of 99.3%and a yield of 473 mg of icaritin from 1.1 g icariin.

Identification of Icaritin Metabolites in Rats by LC-MS/MS

Objective Icaritin is the main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese materia medica Epimedii Herba. Modern pharmacological studies have demonstrated that icaritin has a wide range of biological activities. However, its metabolites and biotransformation pathways have not yet been comprehensively investigated. The present study aims to identify icaritin metabolites in rats by using a sensitive and effective LC-MS/MS method. Methods The plasma and urine samples of rats were collected before （blank） and after oral administration of icaritin, and subjected to liquid-liquid extraction with ethyl acetate. The full-scan LC-MS chromatograms of the plasma and urine samples were compared with those of blank samples to detect the possible metabolites, which were later detected by their product ion spectra. Results A total of 23 metabolites were identified, and conjugated icaritins produced by glucuronidation, glycosylation, and sulfation were its major metabolites. Minor demethylation, hydrogenation, and oxidation metabolites were also found. Conclusion Phase II metabolism is the main metabolic pathway of icaritin.

Icaritin Attenuates Lipid Accumulation by Increasing Energy Expenditure and Autophagy Regulated by Phosphorylating AMPK

Background and Aims:Lipid accumulation is the major characteristic of non-alcoholic fatty liver disease,the prevalence of which continues to rise.We aimed to investigate the effects and mechanisms of icaritin on lipid accumulation.Methods:Cells were treated with icaritin at 0.7,2.2,6.7,or 20μM for 24 h.The effects on lipid accumulation in L02 and Huh-7 cells were detected by Bodipy and oil red O staining,respectively.Mitochondria biogenesis of L02 cells was detected by MitoTracker Orange staining.Glucose uptake and adenosine triphosphate content of 3T3-L1 adipocytes and C2C12 myotubes were detected.The expression levels of proteins in the adenosine 5′-monophosphate-activated protein kinase(AMPK)signaling pathway,biomarkers of autophagy,and mitochondria biogenesis were measured by western blotting.LC3 puncta were detected by immunofluorescence.Results:Icaritin significantly attenuated lipid accumulation in L02 and Huh-7 cells and boosted the mitochondria biogenesis of L02 cells.Icaritin enhanced glucose uptake,decreased adenosine triphosphate content,and activated the AMPK signaling pathway in 3T3-L1 adipocytes and C2C12 myotubes.Icaritin boosted autophagy and also enhanced the initiation of autophagic flux in 3T3-L1 preadipocytes and C2C12 myoblasts.However,icaritin decreased autophagy and promoted mitochondria biogenesis in 3T3-L1 adipocytes and C2C12 myotubes.Conclusions:Icaritin attenuates lipid accumulation by increasing energy expenditure and regulating autophagy by activating the AMPK pathway.

Metabolic profiling of icaritin in rats using UHPLC-Q/TOF-MS

Objective: To identify the in vivo metabolites of icaritin and speculate its metabolic profiling in rats.Methods: The plasma, bile, urine, and feces of rats were collected after orally administration of icaritin at a dose of 100 mg/kg and detected by an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS/MS) in both positive and negative modes.The data of treated and control groups were compared and analyzed with the aid of Metabolynx XS software.Results: A total of 25 metabolites were identified in the biosamples, and 14 of them were reported for the first time to our knowledge.Conclusion: The main metabolite types of icaritin in rats were glucuronide conjugation, methylation, hydroxylation, reduction, and acetylation.

Protective effect of icaritin on focal cerebral ischemic–reperfusion mice

Objective： To investigate the protective effects of icaritin （ICT）, one of the active ingredients in Epimedii Folium, on mouse model of cerebral ischemia-reperfusion （I/R） in vivo. Methods： ICR mice were subjected to an I h transient middle cerebral artery occlusion （MCAO） and fol- lowed by 24 h of reperfusion. Neurological deficits, infarct volume, brain edema and survive rate were measured, respectively. The levels of brain IL-1β, TNF-a, ROS and DNA-binding activity of NF-KB p65 were measured by ELISA kits. The levels of malondialdehyde （MDA） and activities of superoxide dismu- tase （SOD） were detected by spectrophotometry, and the release of nitric oxide （NO） were detected by Griess kit. Results： ICT markedly reduced the neurological deficit scores, brain edema, infarct volume and increased the survival rate of the cerebral I/R mice. The expression of IL-Iβ, TNF-α, NO, MDA and DNA-binding activity of NF-KB p65 were significantly inhibited by ICT, while the activity of SOD were up-regulated at the same time. Conclusion： ICT possessed significant neuroprotective effects in cerebral I/R mice, which might be related to prevent neuroinflammatory and oxidative damage.

Improved Synthesis of Icaritin and Total Synthesis of β-Anhydroicaritin

Natural products icaritin and β-anhydroicaritin with P-glycoprotein(P-gp) inhibitory activities were ciently synthesized in nine steps from commercially available phloroglucinol. A modified Algar-Flynn-Oyamada cyclization and relay Claisen-Cope rearrangement were employed in this concise route. Oiir synthesis offers opportunities to synthesize various icariin analogues for biological and pharmacological investigations.

淫羊藿素通过调控IL-6/JAK2/STAT3途径抑制卵巢癌的发生与恶性生长

目的:探讨淫羊藿素通过调控IL-6/JAK2/STAT3信号通路对卵巢癌A2780和SKOV3细胞增殖、凋亡和侵袭及对小鼠卵巢癌移植瘤模型的影响。方法:将人卵巢癌A2780和SKOV3细胞置于细胞培养箱中培养,设置空白对照组和淫羊藿素低、中、高剂量组(5.00、10.00、20.00μmol/L),药物干预48 h。采用克隆形成实验检测细胞增殖,流式细胞术检测细胞凋亡,Transwell实验检测细胞侵袭。Western blotting检测Ki67、PCNA、cleaved Caspase-3、cleaved Caspase-9、E-cadherin、N-cadherin、VEGF、IL-6、p-JAK2和p-STAT3蛋白相对表达量。建立裸鼠卵巢癌移植瘤模型,检测移植瘤质量和体积,采用Western blotting检测Ki67、cleaved Caspase-3、E-cadherin、N-cadherin、VEGF、IL-6、p-JAK2和p-STAT3蛋白的表达量。结果:与空白对照组比较,淫羊藿素中剂量和高剂量治疗后,卵巢癌A2780和SKOV3细胞增殖、侵袭降低,凋亡增加,体内外Ki67、PCNA、cleaved Caspase-3、cleaved Caspase-9、N-cadherin、VEGF、IL-6、p-JAK2和p-STAT3蛋白的表达量下调,E-cadherin蛋白的表达量上调。结论:淫羊藿素可抑制卵巢癌A2780和SKOV3细胞的增殖和侵袭,促进细胞凋亡,其机制与淫羊藿素影响IL-6、JAK2和STAT3的表达有关。

Icaritin enhances sorafenib-induced apoptosis through a mitochondria-dependent pathway

Sorafenib remains the standard systemic treatment for advanced human hepatocellular carcinoma(HCC).However,the low response rate,high recurrence,and high progression limit the therapeutic efficacy.Therefore,a combination therapy strategy was advanced to strengthen the antitumor effects of sorafenib.In the present study,we aimed to evaluate whether icaritin could enhance the inhibitory effects of sorafenib on HCC cells and clarify the underlying mechanism.The cell viability was evaluated via MTT assay,and the synergistic inhibitory effects of sorafenib and icaritin were verified by calculating the combination index(CI).Their combined effects on cell proliferation or apoptosis were investigated using colony formation assay and flow cytometry.Mitochondrial membrane potential(MMP)was detected by flow cytometric assay.The protein expressions associated with the apoptotic pathway were determined by Western blotting analysis.The data demonstrated that sorafenib and icaritin exerted synergistic inhibitory effects on cell viability(CI<1).Icaritin enhanced the inhibitory effect of sorafenib on colony formation and sorafenib-induced apoptosis of HCC cells.We discovered a reduced level of antiapoptotic Bcl-2 and an elevated level of proapoptotic protein Bax when the cells were exposed to the combination.The effect of cleaved and activated PARP was also enhanced.Cleaved caspase-9 and cleaved caspase-3 were increased markedly in the combination group.Furthermore,the combination of icaritin and sorafenib significantly increased the loss of MMP compared with the single treatment group and induced the release of cytochrome c from the mitochondria to the cytosol.These findings indicated that icaritin could enhance sorafenib-induced cytotoxicity and trigger sorafenib-induced apoptosis through a mitochondria-dependent pathway.