6,8-二-三氟甲基-7-乙酰基白杨素抗肝癌作用实验研究

目的:研究6,8-二-三氟甲基-7-乙酰基白杨素(dFMAChR)体内外抗肝癌作用。方法:MTT比色法测定dFMAChR对体外培养人肝癌细胞Hep G2细胞和人胚肝细胞L-02细胞增殖的影响;平皿克隆形成法及软琼脂克隆形成法测定dFMAChR对体外培养Hep G2细胞的锚定依赖性及非锚定依赖性生长作用;PI染色流式细胞术(FCM)分析dFMAChR对Hep G2细胞周期的影响;人肝癌裸鼠异种移植瘤模型治疗实验评价dFMAChR治疗人肝癌的有效性。结果:dFMAChR抑制体外培养人肝癌Hep G2细胞增殖和生长,其效价强度高于先导化合物白杨素(ChR),而对人胚肝(L-02)细胞的毒性小,其选择指数为42.96。PI染色FCM结果显示3.0μM,10.0μM,30.0μM的dFMAChR作用于Hep G2细胞48h后,其G1期累积细胞百分率分别为64.5%、69.1%、78.4%,较溶媒对照组和ChR 30.0μM的58.2%和68.3%有显著提高,呈现G1期阻滞现象。人肝癌裸鼠异种移植瘤模型治疗实验结果显示:dFMAChR对人肝癌异种移植瘤生长具有显著抑制作用,20,40,80 mg/kg的dFMAChR对移植瘤的瘤重抑制率分别为40.17%,47.41%和66.81%。结论:dFMAChR具有人肝癌治疗作用。     

6,8-二-三氟甲基-7-乙酰氧基白杨素抑制酪蛋白激酶诱导人卵巢癌CoC1细胞凋亡

目的:研究6,8-二-三氟甲基-7-乙酰氧基白杨素(9dFMAChR))抑制体外培养人卵巢癌细胞系CoC1细胞增殖和诱导凋亡作用及机制。方法:体外培养CoC1细胞,MTT比色法测定dFMAChR对CoC1细胞增殖活性的影响;AO/EB染色法荧光显微镜观察dFMAChR诱导CoC1凋亡细胞的形态学改变;DNA凝胶电泳确证dFMAChR诱导CoC1细胞凋亡作用;Western blotting法分析dFMAChR对CoC1细胞酪蛋白激酶CK2α蛋白表达和活性的影响。结果:MTT比色法结果显示,dFMAChR有效抑制CoC1细胞增殖活性,呈剂量依赖性;其IC50值为11.8μM。AO/EB染色荧光显微镜观察dFMAChR处理后,部分CoC1细胞呈现典型凋亡细胞形态特征;dFMAChR(10.0μM)处理CoC1细胞48 h,琼脂糖凝胶电泳出现“梯形”DNA条带。Western blotting分析结果表明:dFMAChR下调酪蛋白激酶CK2α表达,呈浓度和时间依赖性。结论:dFMAChR具有抑制人卵巢癌CoC1细胞增殖和诱导细胞凋亡作用;作用机制与其抑制酪蛋白激酶CK2α表达有关。     

Antiviral Activity of Chrysin Derivatives against Coxsackievirus B3 in vitro and in vivo

Chrysin is a 5,7-dihydroxyflavone and was recently shown to potently inhibit enterovirus 71 (EV71) by suppressing viral 3C protease (3C^pro) activity. In the current study, we investigated whether chrysin also shows antiviral activity against coxsackievirus B3 (CVB3), which belongs to the same genus (Enterovirus) as EV71, and assessed its ability to prevent the resulting acute pancreatitis and myocarditis. We found that chrysin showed antiviral activity against CVB3 at 10 μM, but exhibited mild cellular cytotoxicity at 50 μM, prompting us to synthesize derivatives of chrysin to increase the antiviral activity and reduce its cytotoxicity. Among four 4-substituted benzyl derivatives derived from C(5) benzyl-protected derivatives 7, 9–11 had significant antiviral activity and showed the most potent activity against CVB3 with low cytotoxicity in Vero cells. Intraperitoneal injection of CVB3 in BALB/c mice with 1×10⁶ TCID₅₀ (50% tissue culture infective dose) of CVB3 induced acute pancreatitis with ablation of acinar cells and increased serum CXCL1 levels, whereas the daily administration of 9 for 5 days significantly alleviated the pancreatic inflammation and reduced the elevation in serum CXCL1 levels. Collectively, we assessed the anti-CVB3 activities of chrysin and its derivatives, and found that among 4-substituted benzyl derivatives, 9 exhibited the highest activity against CVB3 in vivo, and protected mice from CVB3-induced pancreatic damage, simultaneously lowering serum CXCL1 levels.     

Assessment of ultra-structure,viability and function of lipopolysaccharides-stimulated human dermal fibroblasts treated with chrysin and exosomes (in vitro study)

BackgroundLipopolysaccharides (LPS) stimulate production of inflammatory cytokines. Chrysin is flavonoid beneficial for treatment of inflammatory conditions. Bone marrow mesenchymal stem cell (BM-MSC) exosomes have regenerative ability in different tissues.ObjectiveTo assess potential role of chrysin and BM-MSC exosomes on ultra-structure, viability and function of human dermal fibroblasts-adult (HDFa) stimulated by LPS.MethodsHDFa cells were divided into: Group I: Cells received no treatment. Group II: Cells were stimulated with LPS. Group III: LPS stimulated cells were treated with chrysin. Group IV: LPS stimulated cells were treated with exosomes.ResultsAfter 48 h, ultrastructural examination of HDFa cells in Group I revealed intact plasma membrane and numerous cytoplasmic organelles. Group II displayed destructed plasma membrane and apoptotic bodies. Group III showed intact plasma membrane with loss of its integrity at some areas. Group IV demonstrated intact plasma membrane that showed fusion with exosomes at some areas. Statistical analysis of MTT represented highest mean value of cell viability% in Group IV followed by Groups III, I and II respectively. Statistical analysis of enzyme-linked immunosorbent assay (ELISA) showed the highest mean value of interleukin-1β (IL-1β) was in Group II followed by Groups III, IV and I, while highest mean values of interleukin-10 (IL-10), nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins were in Group I, followed by Groups IV, III and II respectively.ConclusionsLPS have harmful consequences on ultra-structure, viability and function of HDFa cells. BM-MSC exosomes have better regenerative action on inflamed fibroblasts in comparison to chrysin.     

蜂胶毛胶中总黄酮、高良姜素和白杨素含量分析

目的：建立蜂胶毛胶中总黄酮以及高良姜素、白杨素含量测定方法,测定不同基原毛胶中上述各指标的含量。方法：以芦丁为对照品,采用uV法测定总黄酮含量,检测波长为415nm;用HPLC同时测定高良姜素、白杨素含量,采用Welchrom C18柱,以甲醇-0．15％磷酸溶液梯度洗脱,检测波长为268nm。结果：总黄酮在0～1．248mg线性关系良好,r=0．9999（n=6）,平均回收率为98．82％,RSD=1．45％。高良姜素、白杨素分别在0．089～0．4450,0．115～0．580μg线性关系良好,r均达到1．000,平均回收率分别为99．03％、99．32％,RSD分别为1．10％、1．72％。结论：该方法准确、可靠、简便易行,可用于蜂胶毛胶的定量分析,为蜂胶毛胶、其提取物以及蜂胶制品的质量评价和控制提供了依据。     

8-硝基白杨素对人宫颈癌Hela细胞增殖和凋亡的影响

目的：观察8-硝基白杨素(8-NOChR)抑制体外培养人宫颈癌Hela细胞增殖和诱导凋亡作用。方法：体外培养人宫颈癌Hela细胞系细胞。MTT比色法测定Hela细胞增殖活性。软琼脂培养克隆形成法检测Hela细胞集落形成能力。AO/EB染色荧光显微镜观察Hela细胞凋亡形态学改变。DNA凝胶电泳观察梯形DNA条带。结果：MTT比色测定显示,8-NOChR抑制Hela细胞增殖,呈剂量依赖性。软琼脂培养克隆形成法检测表明,8-NOChR显著抑制Hela细胞集落形成,呈剂量依赖性。AO/EB染色荧光显微镜观察发现,8-NOChR诱导Hela细胞呈现典型凋亡细胞形态特征。8-NOChR（30μmol/L）处理Hela细胞72h,琼脂糖凝胶电泳出现“梯形”DNA条带。结论：8-NOChR具有抑制人宫颈癌Hela细胞增殖和诱导细胞凋亡作用     

白杨黄素影响两种胃癌细胞系增殖与凋亡的研究

 目的 研究白杨黄素对两种胃癌细胞系SGC-7901和MKN-45是否具有抑制作用的分子生物学机制.方法 通过MTT法检测白杨黄素对SGC-7901和MKN-45细胞的增殖抑制作用,流式细胞术测定经白杨黄素处理后两种细胞的细胞周期,Westen-blot测定凋亡相关因子Caspase-3、Survivin和Bcl-2的表达水平.结果 白杨黄素对SGC-7901和MKN-45细胞具有增殖抑制作用,呈时间浓度依赖性;能够将细胞周期阻滞于Sub G1期;能上调Caspase-3,下调Survivin和Bcl-2的基因表达.结论 白杨黄素能够诱导SGC-7901和MKN-45细胞的凋亡并有效地抑制其增殖.其机制可能与干扰细胞周期及激活Caspase-3,抑制Survivin和Bcl-2有关.     

Chrysin promotes tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induced apoptosis in human cancer cell lines

Chrysin exists widely in plants, honey and propolis. The anti-cancer property of chrysin has been demonstrated though the molecular mechanism is not clear. In this study, we found that pre-treatment with chrysin could promote the cell death induced by TRAIL according to the morphological changes and appearance of sub-G1 peak in four human cancer cell lines. In HCT-116 cells, the results of flow cytometry analysis showed that the percentage of sub-G1 reached (38.89 ± 3.78) % when pre-treatment of chrysin was used at 40 μM, but that was only (2.53 ± 0.10) % in the untreated group and (13.22 ± 0.20) % in TRAIL alone group. The differences between the combination and the untreated or TRAIL alone group were all significant (P < 0.05) and dose-dependent effect was obvious. Similar results were obtained in CNE1 cells. In the search of molecular mechanisms, we found that pre-treatment with chrysin could increase TRAIL-induced degradation of caspase 3, caspase 8, PARP proteins. Z-VAD-fmk, which is a pan-caspase inhibitor, could inhibit the apoptosis enhanced by the combination of chrysin and TRAIL. All data indicate that chrysin can enhance the apoptosis induced by TRAIL, and the apoptosis is caspase-dependent and related to the activation of caspase 8.     

白杨素和它的四乙基二磷酸酯对人宫颈癌 Hela细胞增殖和核基质蛋白影响的研究

探讨白杨素和它的四乙基二磷酸酯对人宫颈癌细胞的增殖、分化和核基质蛋白的影响。用终浓度为10μmol/L的白杨素(chrysin,CR)和四乙基白杨素二磷酸酯(tetraethyl bis-phosphoric ester of chrysin,CP)分别处理人人宫颈癌Hela细胞,记录细胞生长数目,进行增殖分化型细胞染色,测定软琼脂集落形成率,应用SDS-PAGE技术分析细胞核基质蛋白成分的变化。CR/CP能明显抑制Hela细胞的增殖,处理组与对照组之间差异有显著性(P<0.05);处理组与对照组比较增殖型细胞减少,分化型细胞数增加,双层软琼脂中细胞集落形成受到明显抑制,对照组集落普遍比药物组集落大,且单个集落中细胞数目多(P<0.05);通过Quantity One软件分析,实验组(CR组与CP组)与对照组(C组)比较,相对分子质量70 0006、8 000、13 000为增加的条带,14 000、17 000、18 000为增强的条带,58 000、43 000是减弱的条带。两实验组相比,以CP组变化显著。CR和CP对Hela细胞具有增殖抑制和诱导分化作用。同时还可以诱导Hela细胞核基质蛋白成分改变,这可能对肿瘤细胞基因表达调控、分化及凋亡起重要作用。     

The Protective Effect of Chrysin Against Cisplatin İnduced Ototoxicity in Rats

Ototoxicity is a common side effect of cisplatin chemotherapy. The aim of this study was to investigate the potential protective effect of chrysin against cisplatin-induced ototoxicity. Thirty-four adult female Wistar albino rats were separated into four groups: a cisplatin group (Group A), with cisplatin administered to ten rats once daily for three consecutive days at doses of 8 mg/kg body weight intraperitoneally (i.p.); a cisplatin plus chrysin group (Group B), with 8 mg/kg of cisplatin administered i.p. daily to ten rats for three consecutive days and 25 mg/kg of chrysin administered via oral gavage in a corn oil for 5 days: a chrysin group (Group C), with 25 mg/kg of chrysin administered via oral gavage in corn oil for 5 days to seven rats; and a control group (Group D), with 5 ml/kg of corn oil administered to seven rats via oral gavage for 5 days. Distortion product otoacoustic emission measurements were performed in the same ear of the rats under general anesthesia at baseline and on the first and fifth days after drug administration. No significant differences were noted between the measurements either in the chrysin group or in the control group. In the cisplatin group, there was a significant worsening of hearing compared to baseline and the measurements on the fifth day at all frequencies. In the statistical analysis, a statistically significant difference was observed at 5039, 6351, 8003, and 10078 Hz frequencies between the measurements on the first and fifth days. In the cisplatin plus chrysin group, there were statistically significant differences at frequencies of 2,003 and 5,039 Hz between the measurements at baseline and on the fifth day, at 3,175 and 5,039 Hz between the measurements on the first and fifth days, and at 8,003 and 100,078 Hz between the measurements at baseline and on the first day. According to these results, this study demonstrates that cisplatin-related ototoxicity can be prevented in rats by the administration of chrysin.