EWS Knockdown and Taxifolin Treatment Induced Differentiation and Removed DNA Methylation from p53 Promoter to Promote Expression of Puma and Noxa for Apoptosis in Ewing’s Sarcoma

Ewing’s sarcoma is a pediatric tumor that mainly occurs in soft tissues and bones. Malignant characteristics of Ewing’s sarcoma are correlated with expression of EWS oncogene. We achieved knockdown of EWS expression using a plasmid vector encoding EWS short hairpin RNA (shRNA) to increase anti-tumor mechanisms of taxifolin (TFL), a new flavonoid, in human Ewing’s sarcoma cells in culture and animal models. Immunofluorescence microscopy and flow cytometric analysis showed high expression of EWS in human Ewing’s sarcoma SK-N-MC and RD-ES cell lines. EWS shRNA plus TFL inhibited 80% cell viability and caused the highest decreases in EWS expression at mRNA and protein levels in both cell lines. Knockdown of EWS expression induced morphological features of differentiation. EWS shRNA plus TFL caused more alterations in molecular markers of differentiation than either agent alone. EWS shRNA plus TFL caused the highest decreases in cell migration with inhibition of survival, angiogenic and invasive factors. Knockdown of EWS expression was associated with removal of DNA methylation from p53 promoter, promoting expression of p53, Puma, and Noxa. EWS shRNA plus TFL induced the highest amounts of apoptosis with activation of extrinsic and intrinsic pathways in both cell lines in culture. EWS shRNA plus TFL also inhibited growth of Ewing’s sarcoma tumors in animal models due to inhibition of differentiation inhibitors and angiogenic and invasive factors and also induction of activation of caspase-3 for apoptosis. Collectively, knockdown of EWS expression increased various anti-tumor mechanisms of TFL in human Ewing’s sarcoma in cell culture and animal models.

Fabrication of taxifolin loaded zein-caseinate nanoparticles and its bioavailability in rats

Taxifolin loaded zein-caseinate nanoparticles(TZP)were fabricated by the anti-solvent method and were used as an oral delivery vehicle to improve their bioavailability in the rat.The formulations of TZP were optimized.With mass ratio of 1:1:2 between taxifolin,zein and sodium caseinate,the particle size andζpotential of TZP were(168.74±0.35)nm and−(57.67±0.25)mV,while the encapsulation and loading efficiency of taxifolin were(85.83±0.89)%and(17.11±0.88)%,respectively.After freeze-drying,TZP exhibited excellent redispersibility in water without aggregation.Physicochemical characterization showed that taxifolin existed in amorphous form in TZP and its interaction with the protein was observed.After encapsulating in TZP,the excellent dispersion of taxifolin in water signifi cantly improve its diffusion velocity through a semipermeable membrane.After oral administration,taxifolin and its 5 metabolites were identifi ed in rat plasma by ultra high performance liquid chromatography(UPLC)with quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS).The dynamic variation of taxifolin and its metabolites in plasma were then quantifi ed by UPLC with a triple-quadrupole typemass spectroscopy(UPLC-QqQ-MS/MS).A pharmacokinetic study showed that the bioavailability of taxifolin increased from 0.35%to 0.52%through TZP fabrication.The plasma concentration of taxifolin glucuronide and methylated taxifolin glucuronide was much higher than taxifolin.Glucuronidation was the dominating metabolism pathway of taxifolin in vivo.

Taxifolin attenuates ischemia-reperfusion induced oxidative ovarian damage in rats

Objective:To investigate preventive effects of taxifolin on ischemia-reperfusion induced oxidative ovarian damage in rats.Methods:A total of 18 female Wistar albino rats were randomly and equally divided into three groups:the sham group,the ovarian ischemia reperfusion group,and the 50 mg/kg taxifolin+ovarian ischemia reperfusion group.The ovarian ischemia reperfusion and taxifolin+ovarian ischemia reperfusion groups were exposed to ischemia for 2 h and then followed by two-hour reperfusion protocol.Biochemical and histopathologic examinations were performed on the extracted ovaries.Results:Levels of malondialdehyde and cyclooxygenase-2 were increased,while reduced-glutathione and cyclooxygenase-1 were decreased in the ovarian ischemia reperfusion group.However,these values were reversed in the taxifolin+ovarian ischemia reperfusion group.Similarly,the number of primordial and developing follicules decreased in the ovarian ischemia reperfusion group,while they were within normal range in the taxifolin+ovarian ischemia reperfusion group.Conclusions:Ischemia followed by reperfusion leads to oxidative stress-related ovarian injury,and taxifolin may be useful for protecting ovarian tissue from such injury.

Effect of taxifolin on cold-shock damages in spermatozoa in rabbits

Objective:To investigate the effect of taxifolin added to rabbit semen on freezing-induced cold-shock damages in spermatozoa.Methods:Semen was collected from six adult New Zealand rabbits once a week by artificial vagina.The collected semen was pooled at 38℃ and divided into four equal volumes.They were diluted with 0,50,100 and 200μM taxifolin-containing Tris+egg yolk extender at 38℃ and their temperatures were lowered to 4℃.Following equilibration,semen drawn into 0.25 mL straws were frozen in an automatic semen freezing device and stored in liquid nitrogen container at-196℃.Samples were thawed in 38℃ water for 25 s and the analyses of motility,kinematic parameters,morphological deformities,changes in membrane integrity,mitochondrial membrane potential,dead-live ratio,acrosomal damages and as well as oxidative stress analyses were performed in semen.Results:Addition of 50μM taxifolin significantly improved motility(total,progressive,rapid and static),high mitochondrial membrane potential and the ratios of spermatozoa with acrosomal damage compared to the control group.Compared to the control group,malondialdehyde(MDA)levels in the 50 and 100μM taxifolin groups were significantly lower,while the MDA level was high and viable spermatozoa ratio was low in the 200μM taxifolin group.There were no significant differences between the groups in terms of kinematic parameters,morphological deformities,membrane integrity and antioxidant levels.Conclusions:The low dose of taxifolin(50μM)has a positive effect and the high dose(200μM)has a negative effect.Therefore,it is concluded that the addition of low-dose(50μM)taxifolin to the extenders would be a useful additive in reducing cold-shock damage that occurs during freezing of rabbit semen.

Taxifolin和EGCG对IFN-γ诱导的HaCaT细胞iNOS表达的影响

紫杉叶素和L-(-)-表没食子儿茶素没食子酸酯(EGCG)均属于类黄酮物质,具有抗炎活性.本文采用激光共聚焦显微镜和流式细胞仪检测SMT(iNOS抑制子)对800U/mL IFN-γ诱导的iNOS表达的影响,结果发现人表皮角质细胞HaCaT中800U/mL IFN-γ诱导iNOS蛋白的表达显著提高,同时SMT可以抑制IFN-γ诱导iNOS蛋白的表达.分别用40μg/mL紫杉叶素和40μg/mL EGCG预处理HaCaT细胞,然后用800U/mL IFN-γ诱导HaCaT细胞,结果发现紫杉叶素和EGCG均可以抑制iNOS蛋白的表达,并且EGCG的效果更好.因此,单体类黄酮紫杉叶素和EGCG可用于皮肤炎症的治疗,并在相同浓度下EGCG效果更强.

Taxifolin protects hypoxia-induced cardiomyocytes injury via HIF1-a/HO-1/autophagy pathway

Background Autophagy, a dynamic and efficient process of self-digestion in vivo, has been proven to be ben- eficial for cardiac protection during MI process via removing the additional protein and damaged organelles in the heart. Taxifolin （Tax）, a common plant flavonoid, has been widely used for the treatment of myocardial infarction. However, the underlying mechanism of Tax is largely unknown. Methods Murine arterial cardiomyocytes HL-1 cells were pretreated with Tax and then exposed to hypoxia environment. CCK-8 was performed for the de- termination of cell viability. Monodansylcadaverine （MDC） and Microtubule-associated protein 1A/1B-light chain 3 （LC3） were analyzed by immunofluorescence staining. The relative gene expressions of Hypoxia Inducible Factor-1 （HIFI-a） and Heine Oxygenase-1 （HO-1） were determined by qRT-PCR. Results Tax at 100μm for 6 h has the maximal effect to avoid reducing the cell viability excessively and significantly abolished hypoxia-induced cell death. Both of the MDC and LC3 immunofluorescence staining revealed markedly increase of expression of autophagy with pretreatment of Tax. Finally, qRT-PCR showed the upregulation of HIFI-a and HO-1 with Tax. Conclusions Taken together, Tax might be a potential candidate for the treatment of MI by promoting cardio- myocyte autophagy via the activation of HIF 1-a and HO-1.

Taxifolin attenuates inflammation via suppressing MAPK signal pathway in vitro and in silico analysis

Objective:Taxifolin is a natural flavonoid compound that can be isolated from onions,grapes,oranges and grapefruit.It also acts as a medicine food homology with extraordinary antioxidant and antiinflammatory activity.This study aims to explain the protective effects and potential mechanisms of taxifolin against inflammatory reaction.Methods:Levels of interleukin(IL)-6,IL-1βand intracellular reactive oxygen species(ROS)were assessed in different time after the treatment of taxifolin in RAW264.7 cells induced by lipopolysaccharide(LPS).Subsequently,the m RNA and protein levels of inducible nitric oxide synthase(i NOS),vascular endothelial growth factor(VEGF),cyclooxygenase(COX)-2,tumor necrosis factor(TNF)-a and the phosphorylation expression levels of the MAPK signal pathway were also evaluated.A silico analysis was used to explain the binding situation for the investigation of taxifolin and MAPK signal pathway.And then MAPK inhibitors were used to reveal the expression level of i NOS,VEGF,COX-2 and TNF-a in RAW264.7 cells.Results:It was demonstrated that cell inflammatory damage induced by LPS was significantly alleviated after the treatment of taxifolin.Then,the m RNA and protein levels of i NOS,VEGF,COX-2 and TNF-a were reduced and the phosphorylation expression levels of the MAPK signal pathway were down-regulated remarkably as well.In silico analysis,taxifolin could form a relatively stable combination with MAPK signal pathway.MAPK inhibitors showed increasing or decreasing effect in the m RNA levels of i NOS,VEGF,COX-2 and TNF-a,which suggesting that taxifolin down-regulated i NOS,VEGF,COX-2 and TNF-a expressions were not entirely through the MAPK pathway.Conclusion:This finding demonstrated that taxifolin improved the inflammatory responses that partly involved in the phosphorylation expression level of MAPK signal pathway in RAW264.7 cells exposed to acute stress.

Protective effects of Taxifolin on H_2O_2-induced damage in H9C2 cells

Background Taxifolin（Tax） is an essential natural antioxidant. Multiple studies have shown that Tax can protect cardiomyocytes from ischemia-reperfusion injury. However, the underlying mechanism is still unclear.Methods H9C2 cells were randomly divided into control, H_2O_2 group, Tax pretreatment group（Tax ＋ H_2O_2）;Tax effect group. Cell activity was detected by CCK-8 and the intracellular structure was observed by transmission electron microscopy. Autophagy was determine by Western blotting analysis of Beclin-1, Bcl-2 and PKC.Results Tax pretreatment significantly increased anti-apoptotic protein Bcl-2 and autophagy protein Beclin-1.Expression of PKC was inhibited by Tax. Conclusions Tax pretreatment could protect H9 C2 cells against H_2O_2-induced damage through the Bcl-2 and autophagy pathways.

基于病症模型的经方四妙丸防治高尿酸血症痛风的药效物质探讨

目的探索经方四妙丸防治高尿酸血症痛风的药效物质。方法建立高尿酸血症药理模型,采用超高效液相色谱串联四极杆静电场轨道阱质谱联用技术(UPLC-Q-Exactive-MS)分析四妙丸体内外化学成分;以入血成分为基础,采用网络药理学构建四妙丸调控高尿酸血症痛风“活性成分-靶点-通路”网络;以关键活性成分与高尿酸血症痛风关键靶点尿酸生成酶(XDH),尿酸转运体(ABCG2、GLUT9、OAT1、URAT1)及炎症靶点(PTGS2、TLR2、TLR4)进行分子对接,根据对接结果进行实验验证,探讨四妙丸防治高尿酸血症痛风的关键药效物质。结果UPLC-Q-Exactive-MS法共鉴定四妙丸89个成分,包含74个入血成分,即候选成分;网络药理学构建了“入血成分-靶点-通路”网络,入血成分匹配116个高尿酸靶点和173痛风靶点,涉及调节糖脂代谢、氧化应激、炎症反应、ERK1、ERK2和MAPK级联反应、PI3K信号传导等生物过程;调节血脂与动脉粥样硬化、凋亡、AGE-RAGE、TNF、PI3K-Akt、MAPK、TLRs、JAK-STAT、NF-κB等信号通路,发挥多途径调控高尿酸血症痛风疾病网络的作用。分子对接研究表明小檗碱、黄柏碱、木兰花碱、药根碱、巴马汀、黄柏酮、柠檬苦素、苍术素、花旗松素、白术内酯Ⅲ及β-蜕皮甾酮等成分与尿酸合成酶、尿酸转运体及炎症靶点的亲和力良好,Western Blot验证发现花旗松素具有负调节尿酸盐转运蛋白1(URAT1)表达的作用,其是四妙丸防治高尿酸血症痛风的主要药效物质。结论该研究阐述了四妙丸调控高尿酸血症痛风的主要药效物质,可为四妙丸临床应用、质量评价和标准提升提供科学依据。

花旗松素调节MCP-1/CCR2轴对急性淋巴细胞白血病细胞恶性进展的影响

目的:研究花旗松素(Taxifolin)对急性淋巴细胞白血病(ALL)细胞恶性进展的影响及单核细胞趋化蛋白-1(CCL2)/CC趋化因子受体2(CCR2)轴发挥的作用。方法:体外培养ALL-T淋巴细胞CCRF-CEM;CCK-8法检测不同浓度Taxifolin对CCRF-CEM细胞活力影响,筛选本实验用Taxifolin浓度;将CCRF-CEM细胞分为control组、Taxifolin-L组、Taxifolin-M组、Taxifolin-H组、Taxifolin+rCCL2组;EDU法检测各组CCRF-CEM细胞增殖;流式细胞术检测各组CCRF-CEM细胞凋亡;Transwell检测各组CCRF-CEM细胞侵袭能力;平板克隆形成实验检测各组CCRF-CEM细胞集落形成能力;RT-PCR检测各组CCRF-CEM细胞CCL2、CCR2基因表达水平;Western blot检测各组CCRF-CEM细胞CCL2、CCR2、Bcl-2、Bax、Caspase-3蛋白表达水平。结果:选择25、50、100μmol/L三个Taxifolin浓度进行后续实验;与control组相比,Taxifolin-L、M、H组CCRF-CEM细胞凋亡率、Bax、Caspase-3蛋白表达水平显著升高,CCRF-CEM细胞EDU阳性细胞率、侵袭细胞数量、细胞克隆集落数、CCL2、CCR2基因表达水平、CCL2、CCR2、Bcl-2蛋白表达水平均显著性降低,且呈剂量依赖性(P<0.05);与Taxifolin-H组相比,Taxifolin+rCCL2组CCRF-CEM细胞凋亡率、Bax、Caspase-3蛋白表达水平显著性降低,CCRF-CEM细胞EDU阳性细胞率、侵袭细胞数量、细胞克隆集落数、CCL2、CCR2基因表达水平、CCL2、CCR2、Bcl-2蛋白表达水平均显著性升高(P<0.05)。结论:Taxifolin可能通过抑制CCL2/CCR2轴,从而抑制CCRF-CEM细胞恶性进展。

落叶松中花旗松素的提取、抑菌作用及其与白屈菜红碱联用协同抑菌作用机理

本研究以落叶松为原料,采用超声辅助乙醇热浸提法提取花旗松素。以大肠杆菌和金黄色葡萄球菌分别作为典型供试菌,通过观察细菌形态结构,测定细菌生长量、细胞膜泄漏、抗氧化酶系活性等变化情况,分析花旗松素抑菌效果。最终获得纯度达90%、提取率0.35%的花旗松素,且花旗松素对两种菌株的最小抑菌浓度均为1.2 mg/mL,抑菌率分别为81.12%、83.95%。通过电镜扫描后发现菌体表面被破坏伴有内容物泄漏。在Escherichia coli中,培养至24 h的1/2 MIC、1 MIC和2 MIC实验组OD_(260)测试值是同期对照组的1.18、1.52、1.88倍,检测到胞外β-半乳糖苷酶的相对活性分别为79.15%和70.1%。Bliss独立模型计算生长速率为0.0142,表明花旗松素和白屈菜红碱药物联用后对金黄色葡萄球菌表现为协同抑制作用。药物联用条件下,低浓度对细菌细胞膜破坏明显增强,抑菌效果更加显著,培养上清液中β-半乳糖苷酶活达87.01%,较2 MIC单独作用S.aureus酶活性高16.91%,24 h CAT、SOD和POD酶活较单独使用1 MIC浓度花旗松素对S.aureus作用要更高,分别是其1.28、1.25、1.11倍。研究将为花旗松素作为食品保鲜、防腐剂等产品的开发应用奠定理论基础。

基于SynergyFinder探究花旗松素和葛根素的协同抗氧化能力

为探究花旗松素与葛根素的协同抗氧化能力,将两者分别按照体积比1:1、1:2和2:1比例进行复配,通过DPPH、ABTS和FRAP法测定其体外抗氧化活性,进一步运用SynergyFinder软件进行HSA模型评价协同效果。结果表明,花旗松素和葛根素均具有一定的抗氧化能力,当花旗松素6.25~25μg/mL与葛根素2~25 mg/mL,花旗松素0~6.25μg/mL与葛根素3.12~12.5μg/mL,花旗松素2.5~10μg/mL与葛根素0~250μg/mL三个浓度范围内1:1复配时,分别对DPPH自由基、ABTS阳离子自由基和FRAP的协同抗氧化能力最强,HSA模型评分最高,分别为13.486、8.383和1.930。因此,选用花旗松素与葛根素进行复配使用,在体积比1:1具有较强的协同抗氧化能力。

花旗松素的提取、检测及功能研究进展

花旗松素是一种二氢黄酮类化合物,常见于松科植物和水果蔬菜中。其常见提取方法有乙醇加热提取法、超声波辅助提取法以及闪式提取法。检测花旗松素最常用的方法为高效液相色谱法。体外和动物试验结果证实花旗松素具有抗氧化、抗癌、抗炎、保护肝脏等多种生物活性。本文对花旗松素提取、检测方法以及其抗氧化、抗炎、抗癌作用机制进行了综述,以期为其工业化生产和应用提供参考。

花旗松素磷脂复合物制备优化及其体外模拟消化释放研究

为了提高花旗松素的利用率,该研究制备花旗松素磷脂复合物,采用Box-Behnken响应面试验优化磷脂复合物制备工艺,并进行表征分析验证磷脂复合物的形成;通过平衡溶解度、油水分配系数及模拟消化实验对理化性质和释放度进行研究。结果表明,磷脂复合物最佳制备条件为花旗松素/磷脂质量比1∶3.71、反应溶剂体积12.82 mL、反应时间2.94 h,此条件下的复合率为95.55%。差示扫描量热分析、傅里叶红外光谱、X射线衍射实验表明花旗松素与磷脂间通过氢键作用形成磷脂复合物,且花旗松素由晶型转变为无定形态。磷脂复合物增加了花旗松素在水中的溶解度和水-正辛醇体系中的油水分配系数;体外模拟消化释放实验显示,6 h内磷脂复合物中花旗松素累积释放由48.26%提升至71.86%。磷脂复合物改善了花旗松素的理化性质和体外释放,为花旗松素的高效应用与食品制剂开发提供研究基础。

花旗松素对神经系统疾病的神经保护作用研究进展

花旗松素是存在于松科植物、蔬菜和水果中的天然黄酮类化合物。本文对近年来花旗松素在神经系统疾病中的研究进展进行综述,包括帕金森病、阿尔茨海默病、亨廷顿病、脑淀粉样血管病、脊髓损伤、肝性脑病、脑缺血再灌注、胶质瘤和癫痫,并从花旗松素的抗神经元凋亡、抗神经炎症、抗氧化损伤和抑制蛋白异常积累的神经保护作用机制进行探讨,以期为进一步了解和研究花旗松素对神经系统疾病的防治作用提供科学依据。

花旗松素对人卵巢癌的作用机制

目的基于网络药理学和体外细胞实验探究花旗松素治疗卵巢癌的作用机制。方法查询文献及数据库检索收集花旗松素的作用靶点,采用SuperPred数据库预测靶点,下载疾病基因,找到药物与疾病靶点的交集,进行蛋白质-蛋白质相互作用分析,获取核心靶点,进行富集分析及分子对接;体外培养人卵巢癌SKOV3细胞,设立空白对照组、花旗松素处理组(10、30μg/ml),比较各组细胞抑制率及磷酸化丝氨酸/苏氨酸特异性蛋白激酶(p-AKT)、丝氨酸/苏氨酸特异性蛋白激酶(AKT)表达。结果花旗松素作用于卵巢癌有79个潜在靶点和12个核心靶点。富集分析结果提示,靶点蛋白在癌细胞内异常表达,通过调节代谢等生物过程,影响酶类及蛋白质结合等功能。KEGG主要富集在PI3K/AKT通路,分子对接提示花旗松素与各靶点具有良好的结合活性。体外实验证实,花旗松素可抑制卵巢癌细胞的活性;Western blot结果显示,花旗松素可降低p-AKT/AKT蛋白的表达,从而发挥抗肿瘤作用。结论花旗松素可通过PI3K/AKT通路抑制卵巢癌细胞的增殖。

紫杉叶素调控PI3K/AKT/mTOR通路对糖尿病模型大鼠氧化应激和胰岛功能的机制研究

目的:探索紫杉叶素(TAX)调控磷脂酰肌醇3-激酶(PI3K)/丝苏氨酸蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路对糖尿病模型大鼠氧化应激和胰岛功能的影响。方法:102只无特定病原体(SPF)级SD大鼠高脂高糖饮食饲养4周,建立2型糖尿病大鼠模型,造模成功后随机分为Model组、TAX低剂量组(5 mg/kg)、TAX中剂量组(10 mg/kg)、TAX高剂量组(20 mg/kg)、PI3K激活剂组(0.02 mg/kg PI3K激活剂740Y-P+20 mg/kg TAX)、mTOR激活剂组(10 mg/kg mTOR激活剂MHY1485+20 mg/kg TAX),每组12只。另外取12只正常大鼠作为Control组,造模成功后,给予相应药物,每日1次,连续干预5周。测量各组大鼠体质量;血糖仪检测空腹血糖(FBG);对各组大鼠进行葡萄糖耐量试验(OGTT);酶联免疫吸附测定法(ELISA)测定大鼠血清空腹胰岛素(FINS)水平,计算胰岛素抵抗指数(HOMA-IR)、胰岛β细胞功能指数(HOMA-β)、胰岛素敏感指数(ISI);苏木精-伊红(HE)染色观察大鼠胰腺组织损伤情况,计算胰岛个数;全自动生化分析仪检测大鼠血清总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、游离脂肪酸(FFA)水平;试剂盒检测大鼠血清丙二醛(MDA)、活性氧(ROS)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平;蛋白免疫印迹法(Western Blot)检测大鼠胰腺组织中PI3K/AKT/mTOR通路蛋白表达。结果:与Control组比较,Model组大鼠体质量、FBG、OGTT(0.5、1.0、2.0 h)血糖值、FINS、HOMA-IR、TC、TG、LDL-C、FFA、ROS、MDA、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR增加,ISI、HOMA-β、HDL-C、GSH-Px、SOD、胰岛数量降低(P<0.05);与Model组比较,TAX低剂量组、TAX中剂量组、TAX高剂量组大鼠体质量、FBG、OGTT(0.5、1.0、2.0 h)血糖值、FINS、HOMA-IR、TC、TG、LDL-C、FFA、ROS、MDA、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR水平降低,ISI、HOMA-β、HDL-C、GSH-Px、SOD、胰岛数量增加(P<0.05);PI3K、mTOR激活剂均减弱了高剂量TAX对糖尿病模型大鼠的改善作用(P<0.05)。结论:TAX通过抑制PI3K/AKT/mTOR信号通路,抑制STZ诱导的糖尿病模型大鼠氧化应激,改善胰岛功能。

花旗松素调节AMPK/mTOR信号通路对急性心肌梗死大鼠心肌损伤的影响

目的探讨花旗松素(TAX)调节AMP活化的蛋白质激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对急性心肌梗死(AMI)大鼠心肌损伤的影响。方法2022年2—11月,从90只雄性SD大鼠中按随机数字表法选择18只大鼠作为空白对照(CON)组,其他大鼠构建AMI模型,将造模成功的AMI大鼠按随机数字表法分为AMI组、TAX组(50 mg/kg)、DOR组(0.01 mol/L的AMPK/mTOR抑制剂DOR)、TAX+DOR组(50 mg/kg TAX+0.01 mol/L的AMPK/mTOR抑制剂DOR),每组18只大鼠。观察超声心动图评估舒张期室间隔厚度(IVSd)、舒张期左室内径(LVIDd)、收缩期左室内径(LVIDs)、左心室舒张期后壁厚度(LVPWd)、左室射血分数(EF)和缩短分数(FS);酶联免疫吸附测定(ELISA)检测心肌损伤标志物肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)以及炎症因子白细胞介素(IL)-1β水平;苏木精-伊红(HE)染色检测心脏组织病理变化;2,3,5-氯化三苯基四氮唑(TTC)染色检测心肌梗死面积;TUNEL染色检测细胞凋亡;蛋白质印迹法检测AMPK/mTOR信号通路蛋白水平。结果CON组心脏组织结构完整,细胞排列整齐有序,染色清晰;AMI组出现炎症细胞浸润现象;TAX组炎症细胞浸润现象减轻,而DOR组炎症细胞浸润现象加重;TAX+DOR组与AMI组结构相似。与CON组相比,AMI组LVIDd、LVIDs、CK、CK-MB、LDH、IL-1β含量、细胞凋亡率、梗死面积[(17.82±2.31)%比(3.91±0.15)%]、mTOR(1.26±0.14比0.54±0.04)、核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3(NLRP3)(0.76±0.08比0.26±0.02)蛋白水平显著升高(P<0.05),IVSd、LVPWd、EF、FS以及p-AMPK/AMPK(0.49±0.05比0.93±0.07)蛋白水平显著下降(P<0.05);与AMI组相比,TAX组LVIDd、LVIDs、CK、CK-MB、LDH、IL-1β含量、细胞凋亡率、梗死面积[(12.36±1.76)%比(17.82±2.31)%]、mTOR(0.69±0.07比1.26±0.14)、NLRP3(0.39±0.04比0.76±0.08)蛋白水平显著降低(P<0.05),IVSd、LVPWd、EF、FS以及p-AMPK/AMPK(0.85±0.09比0.49±0.05)蛋白水平显著升高(P<0.05),而DOR组趋势相反(P<0.05);TAX+DOR组减弱了TAX对AMI大鼠心肌损伤的改善作用。结论TAX可通过调控AMPK/mTOR信号通路减轻AMI大鼠心肌损伤。

黄杉素药理学基础及生物合成研究进展

黄杉素是一种二氢黄酮醇类活性天然产物,具有多种显著的药理活性,包括抗氧化,保护心血管和肝脏等,广泛应用于美容、保健、食品和医疗产业。黄杉素目前主要通过植物提取,产量不足限制了其广泛应用,而生物合成方法研究的快速发展有望为其高效获取提供新契机。本文综述了黄杉素的结构性质、生物合成途径、药理作用以及生产方式,并对未来合成生物学技术在黄杉素生产中的应用进行展望。

UPLC法测定水红花子中花旗松素及槲皮素的含量

目的 应用超高效液相色谱法(UPLC)建立同时测定水红花子药材中花旗松素和槲皮素含量的方法。方法 采用Agilent ZORBAX SB-C18色谱柱,以乙腈-0.1%磷酸水为流动相梯度洗脱,柱温25℃,检测波长290 nm,流速0.2 mL·min^(-1),进样量1μL。结果 建立的分析方法方法学考察良好。花旗松素、槲皮素进样浓度分别在3.258~104.272μg·mL^(-1)、2.983~95.452μg·mL^(-1)内呈良好的线性关系(r~2=0.999 9),平均加样回收率为98.8%、98.8%,RSD值为1.9%、2.2%。结论 本方法简便、快速,可在同一波长下快速测定水红花子中花旗松素及槲皮素的含量,为水红花子药材质量控制提供参考依据。