邳州银杏叶黄酮类成分分析

目的：探索银杏叶黄酮类成分之间的相互关系。方法：采用HPLC法(二级管阵列检测器DAD)分别测定了银杏叶中各黄酮的含量。结果与结论：银杏叶中槲皮素和山柰素的含量随树龄的增长而下降，含量的相对关系会发生变化，二、三年树龄叶以槲皮素为主，四年以上树龄叶山柰素所占比例相对较大。     

抑制HSP 70表达对体外培养的脑胶质瘤细胞生长的影响

目的 探讨抑制热休克蛋白７０（ＨＳＰ７０）表达对脑胶质瘤细胞增殖及凋亡的影响。方法 利用细胞培养、电子显微镜及流式细胞仪的方法，观察阻滞ＨＳＰ７０表达后对肿瘤细胞生长的影响。结果 体外培养脑胶质瘤细胞株、经不同浓度槲皮素作用不同时间后，碘化丙碇染色荧光显微镜观察发现细胞核固缩，致密和破裂现象；流式细胞检测发现亚二倍体峰，提示出现细胞凋亡。分析细胞周期，发现凋亡细胞主要出现在Ｇ０、Ｇ１期，随药物的浓     

槲皮素对X射线照射后大鼠免疫功能的影响

目的 探讨槲皮素(QN)对6 Gy X射线照射后大鼠免疫功能及肝脏氧化应激反应的影响。 方法40只大鼠随机分成4组,每组10只,第1组连续7 d 腹腔内注射生理盐水(0.5 ml/150 g体重)作为对照组,第2组(QN组)连续7 d 用槲皮素(40 mg/kg体重)腹腔注射,第3组用单次6 Gy X射线照射,第4组(6 Gy+QN组)连续7 d 腹腔注射槲皮素(40 mg/kg体重),在最后一次注射后1 h,给予单次6 Gy X射线照射。照射后24 h,将大鼠麻醉后断头处死,取其脾脏用四甲基偶氮唑盐(MTT)法检测淋巴细胞转化率,流式细胞术检测CD⁺₄、CD⁺₈及CD⁺₄/CD^＋₈T淋巴细胞的变化,取肝脏检测氧化应激反应并观察一般状态变化。 结果经QN预先处理组的大鼠脾脏淋巴细胞转化率显著高于6 Gy照射组,CD^＋₄、CD⁺₈及CD^＋₄/CD⁺₈ T淋巴细胞百分率亦显著高于单纯照射组(F值分别为8.455、22.644和18.911, P<0.01),尤其以增加CD^＋₄T淋巴细胞百分率为显著。同时发现QN预先处理的照射组大鼠肝脏丙二醛(MDA)含量显著低于照射组(F=10.059, P<0.01),抗氧化酶类超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和抗氧化物质GSH含量显著高于照射组(F值分别为23.688、186.046和19.788, P<0.01)。6 Gy照射组大鼠肝脏肝小叶中心静脉扩张,肝血窦明显充血,并有炎性细胞浸润,而经QN处理后的照射组大鼠肝脏,肝小叶中心静脉轻度扩张,肝血窦充血较6 Gy照射组明显减轻。结论槲皮素对X射线照射后大鼠免疫功能具有明显增强作用,并显著改善了照射大鼠肝脏的氧化应激损伤,对6 Gy照射大鼠起到了一定程度的保护作用。     

Quercetin Confers Tumoricidal Activity Through Multipathway Mechanisms in A N-Methylnitrosourea Rat Model of Colon Cancer

Objective: This research was conducted to explore mechanisms behind the potency of quercetin in inhibiting colon cancer induced in an experimental model. Materials and Methods: Forty adult male rats of Wistar strain were distributed into 4 groups; a negative control group, a colon cancer bearing group, a quercetin-treated group and a 5-fluorouracil (5-FU)-treated group. Serum TAG72 and GAL3 levels were quantified by ELISA. Colonic Wnt5a and Axin-1 gene expression was estimated by PCR. In addition, colonic tissues were subjected to immunohistochemical examination of Bax expression and histological investigation of histopathological alterations. Results: Quercetin elicited significant reduction in serum TAG72 and GAL3 levels, in addition to significant suppression of colonic Wnt5a gene expression and amplification of colonic Axin-1 gene expression. Also, it caused moderate positive reaction for Bax in mucosal epithelium. Conclusion: The present research provides experimental evidence about the activity of quercetin in the colon cancer of rats. Inhibitory effects on cancer development might be ascribable to regulatory action on Wnt signaling and induction of apoptosis.     

槲皮素及其7-OH衍生物对膀胱癌细胞增殖抑制作用及细胞周期的影响

【摘要】 目的 通过细胞试验及裸鼠移植瘤实验,研究槲皮素及其7-OH衍生物对人膀胱癌BIU-87细胞的增殖抑制作用及细胞周期的影响。方法〓体外实验:培养BIU-87细胞,分别加入槲皮素(QUE)及槲皮素单硫酸酯钠盐(SQMS),MTT实验测定并计算生长抑制率及半数抑制浓度IC50;流式细胞仪检测细胞周期。体内实验:观察不同剂量SQNS及槲皮素对BIU-87细胞裸鼠移植瘤生长的抑制作用;瘤体组织切片行HE染色和免疫组化染色检测Ki-67,CyclingB1,CyclingD1蛋白表达情况。结果〓槲皮素及SQMS体外均能抑制BIU-87细胞的增殖,相同浓度两药物间对比差异无统计学意义(P>0.05)。槲皮素使BIU-87细胞周期明显阻滞于G2/M期,而SQMS则主要阻滞于G0/G1期。在体内实验中,中等剂量的SQMS组抑瘤率与槲皮素组相近(P>0.05),CyclingD1表达比槲皮素组下调更明显(P<0.01),而槲皮素组CyclingB1表达比中剂量SQMS组下调更明显(P<0.05)。两组Ki-67表达无显著性差异(P>0.05)。结论〓槲皮素及其衍生物SQMS对膀胱癌细胞BIU-87有抑制增殖作用;中剂量的SQMS对成瘤生长的抑制作用与槲皮素作用相近;槲皮素对人膀胱癌细胞增殖抑制作用可能是通过下调CyclingB1的表达,阻滞细胞周期于G2/M期,而SQMS则可能是通过下调CyclingD1的表达,阻滞细胞周期于G0/G1期。     

Aberrant Epigenetic Alteration in Eca9706 Cells Modulated by Nanoliposomal Quercetin Combined with Butyrate Mediated via Epigenetic-NF-κB Signaling

Since the epigenetic alteration in tumor cells can be reversed by the dietary polyphenol quercetin (Q) orbutyrate (B) with chemopreventive activity, suggesting that Q or B can be used for chemopreventive as well astherapeutic agent against tumors. In this study the polyphenol flavonoid quercetin (Q) or sodium butyrate (B)suppressed human esophageal 9706 cancer cell growth in dose dependent manner, and Q combined with B (Q+B)could further inhibit Eca9706 cell proliferation than that induced by Q or B alone, compared with untreatedcontrol group (C) in MTT assay. The reverse expressions of global DNMT1, NF-κBp65, HDAC1 and Cyclin D1were down-regulated, while expressions of caspase-3 and p16INK4α were up-regulated, compared with the Cgroup in immunoblotting; the down-regulated HDAC1-IR (-immunoreactivity) with nuclear translocation, andup-regulated E-cadherin-IR demonstrated in immunocytochemistry treated by Q or B, and Q+B also displayedfurther negatively and positively modulated effects compared with C group. The order of methylation specific(MS) PCR of p16INK4α: C>B/Q>Q+B group, while the order of E-cadherin expression level was contrary,Q+B>Q/B>C group. Thus, Q/B, especially Q+B display reverse effect targeting both altered DNA methylationand histone acetylation, acting as histone deacetylase inhibitor mediated via epigenetic-NF-κB cascade signaling.     

Neuroprotective potential of Quercetin in combination with piperine against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity

1-Methy-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)is a neurotoxin that selectively damages dopaminergic neurons in the substantia nigra pars compacta and induces Parkinson’s like symptoms in rodents.Quercetin(QC)is a natural polyphenolic bioflavonoid with potent antioxidant and anti-inflammatory properties but lacks of clinical attraction due to low oral bioavailability.Piperine is a well established bioavailability enhancer used pre-clinically to improve the bioavailability of antioxidants(e.g.,Quercetin).Therefore,the present study was designed to evaluate the neuroprotective potential of QC together with piperine against MPTP-induced neurotoxicity in rats.MPTP(100μg/μL/rat,bilaterally)was injected intranigrally on days 1,4 and 7 using a digital stereotaxic apparatus.QC(25 and 50 mg/kg,intragastrically)and QC(25 mg/kg,intragastrically)in combination with piperine(2.5 mg/kg,intragastrically)were administered daily for 14 days starting from day 8 after the 3~(rd) injection of MPTP.On day 22,animals were sacrificed and the striatum was isolated for oxidative stress parameter(thiobarbituric acid reactive substances,nitrite and glutathione),neuroinflammatory cytokine(interleukin-1β,interleukin-6,and tumor necrosis factor-α)and neurotransmitter(dopamine,norepinephrine,serotonin,gamma-aminobutyric acid,glutamate,3,4-dihydroxyphenylacetic acid,homovanillic acid,and 5-hydroxyindoleacetic acid)evaluations.Bilateral infusion of MPTP into substantia nigra pars compacta led to significant motor deficits as evidenced by impairments in locomotor activity and rotarod performance in open field test and grip strength and narrow beam walk performance.Both QC(25 and 50 mg/kg)and QC(25 mg/kg)in combination with piperine(2.5 mg/kg),in particular the combination therapy,significantly improved MPTP-induced behavioral abnormalities in rats,reversed the abnormal alterations of neurotransmitters in the striatum,and alleviated oxidative stress and inflammatory response in the striatum.These findings indicate that piperine can enhance the antioxidant and anti-inflammatory properties of QC,and QC in combination with piperine exhibits strong neuroprotective effects against MPTP-induced neurotoxicity.     

槲皮素对人腺样囊性癌ACC-M细胞增殖和凋亡的影响及机制研究

目的 观察槲皮素对人腺样囊性癌细胞系ACC-M增殖和凋亡的作用,探讨槲皮素影响ACC-M细胞凋亡的可能机制。方法 采用CCK-8 法检测不同浓度的槲皮素对ACC-M细胞增殖的抑制作用并计算抑制率,计算IC₅₀;流式细胞术检测 ACC-M细胞凋亡率和细胞周期分布;实时荧光定量RT-PCR检测 ACC-M细胞中Survivin mRNA的表达。采用SPSS 20.0软件包对数据进行统计学分析。结果 槲皮素抑制 ACC-M细胞增殖的作用明显,且呈浓度依赖性,IC₅₀为151.12 μmol/L（P<0.05）;Annexin V/PI 双染法检测显示,槲皮素能够诱导ACC-M细胞凋亡,且与剂量、时间呈正相关（P<0.05）;PI染色法检测显示,槲皮素主要作用于 G2/M 节点,能将细胞特异性地阻滞于 G2/M 期;RT-PCR检测显示,随着药物浓度增加,ACC-M细胞中Survivin mRNA的表达水平均呈不同程度的降低,200 μmol/L 槲皮素可显著抑制Survivin mRNA的表达。结论 槲皮素能够抑制ACC-M细胞增殖,诱导其凋亡,且呈时间-剂量依赖性。槲皮素可能通过下调Survivin的表达水平而发挥抗腺样囊性癌的作用。     

槲皮素与贯叶连翘提取物合用抗抑郁作用初步研究

目的：比较不同剂量的槲皮素和贯叶连翘提取物（有效成分为贯叶金丝桃素hyperforin,HF）合用的抗抑郁效果,探索二者的协同作用。 方法：雄性ICR小鼠,分为9组,即：空白对照组,阳性对照（帕罗西汀10 mg/kg）组,槲皮素组（A：5 mg/kg,B：10 mg/kg,C：20 mg/kg）,贯叶连翘提取物（HF 10 mg/kg）组,合用组（A：2.5 mg/kg槲皮素+ HF 5 mg/kg;B：5 mg/kg槲皮素+ HF 5 mg/kg;C：10 mg/kg槲皮素+ HF 5 mg/kg）;灌胃给药。用利血平拮抗试验比较各组（n=11~12）拮抗利血平所致低体温、运动不能和低体温的作用,用小鼠悬尾实验比较各组（n=15）动物的悬尾不动时间。 结果：合用B组的利血平拮抗作用和缩短悬尾不动时间的作用与合用C组无显著差异（P>0.05）,其拮抗利血平所致低体温的作用极显著（P<0.01）强于槲皮素B组,其缩短小鼠悬尾不动时间的作用显著（P<0.05）或极显著（P<0.01）强于贯叶连翘提取物（HF 10 mg/kg）组或槲皮素B组。 结论：槲皮素和贯叶连翘提取物在特定剂量比例合用时有协同抗抑郁作用。     

槲皮素在体外哺乳动物细胞中遗传毒性研究

目的研究槲皮素对体外哺乳动物的细胞遗传毒性。方法采用80、40、20、10、5 mg/L5个剂量组的槲皮素在有或无代谢活化条件下处理体外培养的中国地鼠肺成纤维细胞（CHL）3 h后更换新鲜培养液,恢复生长21 h后收获细胞制片,观察槲皮素对哺乳动物细胞染色体的影响。采用200、100、50、25和12.5 mg/L 5个剂量组的槲皮素在有或无代谢活化条件下处理中国仓鼠肺成纤维细胞（V79）3 h后,经过7 d的次黄嘌呤鸟嘌呤磷酸核糖转移酶（HGPRT）突变基因表达和7 d突变基因选择后计数集落形成率并计突变频率,观察槲皮素对哺乳动物HGPRT基因位点的影响。结果在有或无代谢活化条件下槲皮素在浓度〉10 mg/L均能够诱导CHL细胞染色体断裂和交换等,染色体细胞畸变率显著增加（P〈0.01）;而在有或无代谢活化与溶剂对照组相比较槲皮素各剂量组均未发生基因突变频率显著增加（P〉0.05）。结论在体外试验条件下,槲皮素对哺乳动物细胞显示出明显致突变性,存在潜在的遗传毒性。