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.

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.

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.

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.

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.

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。

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。

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.

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。

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.

超高压辅助胶束法提取落叶松中二氢槲皮素的工艺优化

为简化二氢槲皮素提取工艺,降低能耗与成本,提高提取效率,促进二氢槲皮素的综合应用,本研究采用黑龙江省的兴安落叶松为原料,运用超高压辅助胶束提取技术提取落叶松中二氢槲皮素,测定落叶松树根、树干等不同部位的二氢槲皮素总含量。以此总含量为基础,对提取胶束进行筛选,采用响应面试验对提取工艺进行优化,考察了料液比、提取压力、提取次数及胶束浓度4种不同因素对二氢槲皮素提取率的影响,并与微波提取、超声提取、回流提取等不同提取工艺进行能耗与CO_(2)排放比较。结果表明,最终确定提取胶束为茶皂素,最佳提取工艺条件为:茶皂素浓度8%,料液比1:11.5,提取压力157 MPa,提取次数3次,保压时间5min,在此最佳条件下重复进行3次实验,二氢槲皮素实际提取率可达84.35%±1.20%,与预测值84.98%基本一致。与其他提取方法相比超高压辅助胶束提取率最高,单位原料的能耗和CO_(2)排放量均为最低,分别为1.71×10^(-4)kW·h·g^(-1),1.34×10^(-4)kg/g。综上,利用超高压辅助胶束绿色溶剂提取技术提取落叶松中二氢槲皮素,绿色环保、操作简单、工艺合理可靠,并且重复性高,可广泛应用。

皂角刺不同部位高效液相色谱特征图谱及化学模式识别

建立皂角刺药材不同部位高效液相色谱(HPLC)特征图谱及花旗松素含量测定方法,评价皂角刺药材不同部位的质量差异。采用Waters Symmetry(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%甲酸溶液为流动相梯度洗脱,波长为260 nm,柱温为25℃,流量为1.0 mL/min。采用相似度评价、热图聚类分析、主成分分析及方差分析对12批皂角刺药材不同部位的HPLC特征图谱进行分析,并测定花旗松素含量。皂角刺药材不同部位的HPLC特征图谱均确定了8个共有峰,分别指认了香草酸、花旗松素和槲皮素3个特征峰;12批主刺根、支刺根、刺尖HPLC特征图谱与相应对照特征图谱的相似度均大于0.990;热图聚类分析和主成分分析结果可区分主刺根和刺尖;方差分析结果表明峰2、花旗松素和槲皮素是导致皂角刺不同部位质量差异的主要成分;同一皂角刺药材中花旗松素含量均为刺尖>支刺根>主刺根。该法能反映皂角刺不同部位的化学成分差异,对皂角刺药材的质量控制及整体评价具有重要意义。

花旗松素对转化生长因子β1诱导肝星状细胞的激活作用及基因表达谱分析

目的探索花旗松素对肝星状细胞的激活作用,通过转录组测序表征基因表达谱,分析潜在的作用机制。方法在10μg/L转化生长因子β1(transforming growth factor-β1,TGF-β1)诱导下,体外培养大鼠肝星状细胞HSC-T6,分别给予62.5μmol/L、125μmol/L和250μmol/L花旗松素处理,在24 h、48 h分别加入CCK8试剂,450 nm波长测定吸光度,计算不同浓度和时间下的细胞增殖率。在24 h通过AnnexinV/7AAD染色,流式细胞检测细胞凋亡。通过荧光定量聚合酶链式反应(polymerase chain reaction,PCR)法检测α-平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)、Ⅰ型胶原α1链(Collagen1A1)及Ⅲ型胶原α1链(Collagen3A1)基因mRNA相对表达量。通过Western blot检测α-SMA、Collagen1A1蛋白表达量。通过转录组测序,表征细胞基因表达谱并对差异基因进行基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)功能注释及功能富集分析。结果花旗松素可显著抑制肝星状细胞增殖,处理24 h,125μmol/L组和250μmol/L组细胞增殖率分别为(66.7±8.6)%和(45.6±3.0)%。处理48 h,125μmol/L组和250μmol/L组的细胞增殖率分别为(60.5±2.9)%和(43.9±1.9)%,均显著低于模型组(P均<0.05)。花旗松素可促进细胞凋亡,处理24 h,花旗松素62.5μmol/L组、125μmol/L组、250μmol/L组细胞凋亡率分别为(8.537±0.445)%、(8.85±0.169)%及(14.453±0.577)%,均显著高于模型组(P均<0.05)。处理24 h,花旗松素125μmol/L组、250μmol/L组可抑制α-SMA、Collagen1A1、Collagen3A1基因和α-SMA、Collagen1A1蛋白表达(P均<0.05)。差异基因表达表现为促肝纤维化发生相关基因下调,如氧化型低密度脂蛋白受体1(oxidized low-density lipoprotein receptor 1,Olr1)、人从状蛋白(Plexin A4,Plxna4)、反应蛋白1(Spondin 1,Spon1)、聚集蛋白(Agrn)等,以及细胞抗氧化及死亡相关的基因上调,如血红素加氧酶1(heme oxygenase 1,Hmox1)等。结论花旗松素对肝星状细胞具有抑制增殖、促进凋亡、抑制激活的作用,推测其可能通过调控多项基因表达,主要调控因子之间相互作用信号通路、卟啉与叶绿素代谢信号通路、Hedgehog信号通路和PPARγ信号通路发挥作用。

炒水红花子工业化生产参数优化及质量考察

目的优化炒水红花子的工业化生产参数,考察炒水红花子的质量。方法采用正交分析与工业化生产相结合,采用独立性权重法进行综合评分,对炒水红花子工业化生产参数进行优化。以最优炮制工艺,炮制10批不同产地的水红花子,采用高效液相色谱法(HPLC)测定炮制品中花旗松素的含量,薄层色谱法对炮制品进行考察。结果炒水红花子的最优生产工艺为置270℃预热炒锅内,炒制5 min,转速15 r/min。HPLC法测定炒水红花子中花旗松素的含量,进样量在0.04~2.00μg范围内,线性关系良好(r=0.9999),平均回收率为100.3%(RSD=2.3%,n=9),含量结果在0.13%~0.31%之间。薄层色谱法能进行有效鉴别。结论本研究明确了炒水红花子最优工业化生产参数,考察10批不同产地炮制品质量,验证生产工艺稳定可靠。对炒水红花子质量监管和地方标准升级为国家标准提供参考。

花旗松素调控内质网应激PERK-ATF4通路减轻高血压大鼠心肌肥厚的机制研究

目的探讨花旗松素(TAX)对自发性高血压大鼠(SHR)心肌肥厚的影响及分子机制。方法24只SHR分为SHR对照组(SHR组)、TAX组(20 mg/kg)、TAX+PERK激活剂CCT020312(CCT)组(20 mg/kg TAX+2 mg/kg CCT),每组8只;另选8只正常血压Wistar-Kyoto(WKY)大鼠作为正常对照组(WKY组),给予相应的药物持续干预8周。实验过程中观察大鼠血压变化,并于干预结束后超声心动图检测大鼠舒张期室间隔厚度(IVSd)、收缩期室间隔厚度(IVSs)、左心室射血分数(LVEF)判断心肌肥厚程度和心脏功能,计算心脏指数、左心室指数,苏木精-伊红(HE)染色、小麦胚芽凝集素(WGA)染色和Masson染色评估心肌组织病理学变化,实时荧光定量PCR(qRT-PCR)检测心肌组织中心房钠尿肽(ANP)、B型利钠肽(BNP)、I型胶原蛋白α1链(COL1A1)和Ⅲ型胶原蛋白α1链(COL3A1)mRNA表达,Western blot检测心肌组织蛋白激酶R样内质网激酶(PERK)-转录激活因子4(ATF4)通路相关蛋白表达。结果干预结束后,与WKY组相比,SHR组收缩压(SBP)、舒张压(DBP)、IVSd、IVSs、心脏指数、左心室指数、心肌细胞横截面积、胶原容积分数(CVF)、心肌组织ANP、BNP、COL1A1和COL3A1 mRNA表达、葡萄糖调节蛋白78(GRP78)、ATF4、C/EBP同源蛋白(CHOP)蛋白水平和p-PERK/PERK比值升高(均P<0.05),LVEF降低(P<0.05);与SHR组相比,TAX组SBP、DBP、IVSd、IVSs、心脏指数、左心室指数、心肌细胞横截面积、CVF、心肌组织ANP、BNP、COL1A1和COL3A1 mRNA表达、GRP78、ATF4、CHOP蛋白水平和p-PERK/PERK比值降低(均P<0.05),LVEF升高(P<0.05);CCT020312可部分逆转TAX对心脏功能和心肌肥厚的保护作用。结论TAX可通过抑制内质网应激(ERS),改善高血压心肌肥厚,其作用机制可能与抑制PERK-ATF4通路有关。

花旗松素对人宫颈癌Hela细胞的体外抗肿瘤活性及其机理研究

目的观察和探索花旗松素对人宫颈癌Hela细胞的体外抗肿瘤活性及其作用机理。方法结晶紫染色法检测花旗松素对人宫颈癌Hela细胞的增殖抑制作用;RT-PCR法检测与肿瘤细胞凋亡相关基因的表达。结果花旗松素能够抑制人宫颈癌Hela细胞的增殖,呈剂量依赖关系;花旗松素不能诱导Bcl-2 mRNA和Bax mRNA的表达,但能使P53和P21的mRNA表达量增加。结论花旗松素具有良好的体外抑制人宫颈癌Hela细胞增殖作用;花旗松素诱导Hela细胞死亡与细胞凋亡相关,其分子机制可能与升高P53和P21mRNA表达有关,而与Bcl-2和Bax的蛋白转录无关。