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。     

Protective effects of quercetin and chrysin against 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced oxidative stress,body wasting and altered cytokine productions in rats

The aim of this study is to investigate the effects of the quercetin (Q) and chrysin (CH) on oxidative stress, cytokines levels and body weights in rats induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Rats were divided randomly into six equal groups. TCDD, Q and CH were administered by gavages dissolved in corn oil at the doses of 2 µg/kg/week, 20?mg/kg/day and 50?mg/kg/day, respectively. The blood samples were taken from all rats at 60th days to be analyzed for the determination of thiobarbituric acid reactive substances (TBARS), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). The results indicated that although TCDD increased significantly TBARS and TNF-α levels, it caused a decline in the levels of IFN-γ and body weight. In contrast, these effects of TCDD on TBARS, TNF-α, IFN-γ levels and body weight were significantly prevented by treatments of Q and CH. In conclusion, it was determined that TCDD caused the adverse effects on immune functions, body weight and oxidative stress in rats. However, Q and CH administered with TCDD eliminated these adverse effects. These results suggest that Q and CH may play a protective role against TCDD toxicity.     

Influence of chrysin on D-galactose induced-aging in mice: Up regulation of AMP kinase/liver kinase B1/peroxisome proliferator-activated receptor-γ coactivator 1-α signaling pathway

Adenosine monophosphate kinase/liver kinase B1/peroxisome proliferator-activated receptor-γ coactivator 1-α (AMPK/LKB1/PGC1α) pathway has a vital role in regulating age-related diseases. It controls neurogenesis, cell proliferation, axon outgrowth, and cellular energy homeostasis. AMPK pathway also regulates mitochondrial synthesis. The current study evaluated the effect of chrysin on D-galactose (D-gal) induced-aging, neuron degeneration, mitochondrial dysfunction, oxidative stress, and neuroinflammation in mice. The mice were allocated randomly into four groups (10 each group): Group 1: normal control group, Group 2: D-gal group, Groups 3 and 4: chrysin (125 and 250 mg/kg, respectively). Groups 2–4 were injected with D-gal (200 mg/kg/day; s.c) for 8 weeks to induce aging. Groups 3 and 4 were orally gavaged every day concurrent with D-gal. At the end of experiment, behavioral, brain biochemical and histopathological changes were monitored. Chrysin administration elevated discrimination ratio in object recognition, Y Maze percentage alternation, locomotor activity and brain contents of AMPK, LKB1, PGC1α, NAD (P)H quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1), nerve growth factor (NGF) (neurotrophin-3; NT-3), and seretonin as well as reduced brain contents of tumor necrosis factor-alpha (TNF-α), nuclear factor kappa B (NF-κB), advanced glycation end products (AGEs) and glial fibrillary acidic protein (GFAP) compared to D-gal-treated mice. Chrysin also alleviated cerebral cortex and white matter neurons degeneration. Chrysin protects against neurodegeneration, improves mitochondrial autophagy and biogenesis as well as activates antioxidant genes expression. In addition, chrysin ameliorates neuroinflammation and stimulates the release of NGF and serotonin neurotransmitter. So, chrysin has a neuroprotective effect in D-gal induced-aging in mice.     

木蝴蝶树皮中黄芩苷和白杨素的HPLC分析

目的:建立测定小蝴蝶树皮中黄芩苷和白杨素的反相液相色谱法。方法:采用 Phenomenex ODS 3(250 mm×4.6 mm,5μm)色谱柱;流动相:乙睛-0.2%磷酸溶液,梯度洗脱[0～10 min,乙腈-0.2%磷酸溶液比例(25:75);10～25 min 乙腈-0.2%磷酸溶液比例由(25:75)→(60:40)],流速:1.0 mL·min~(-1);检测波长:268 nm;柱温:30℃。结果:黄芩苷、白杨素线性范围分别为0.03～7.55 mg(r=0.9999)和0.01～2.21 mg(r=0.9999);黄芩苷、白杨素的平均回收率(n=9)分别为96.8%和97.2%。结论:该方法简便、准确,重复性好,为木蝴蝶树皮的进一步研究和质量控制提供科学依据。     

ADFMCHR激活PPARγ抑制人肝癌SMMC-7721细胞生长

目的:研究5-烯丙基-7-二氟亚甲基白杨素(ADFMChR)激活过氧化物酶活化因子受体γ(PPARγ)抑制体外培养人肝癌SMMC-7721细胞增殖作用。方法:体外培养人肝癌SMMC-7721细胞。台盼蓝拒染法检测细胞生长;平皿克隆形成法检测细胞锚定依赖性生长;软琼脂克隆形成法检测细胞集落形成能力。结果:细胞计数结果显示,ADFMChR延长SMMC-7721细胞倍增时间,抑制SMMC-7721细胞生长,呈剂量依赖性。但是对预先用PPARγ阻断剂GW 9662孵育30分钟的SMMC-7721细胞,ADFMChR的生长抑制作用明显降低;平皿克隆、软琼脂克隆形成法结果显示,ADFMChR抑制SMMC-7721细胞的锚定依赖性生长能力和锚定非依赖性生长能力,而GW 9662可以降低ADFMChR对SMMC-7721细胞的增值抑制作用。结论:ADFMChR具有抑制人肝癌SMMC-7721细胞生长作用,其机制与激活PPARγ有关。     

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。     

8-硝基白杨素诱导结肠癌HT-29细胞凋亡

目的:观察8-硝基白杨素(NOChR)诱导人结肠癌HT-29细胞凋亡作用。方法:MTT比色法测定细胞增殖活性;流式细胞分析术检测细胞凋亡率;琼脂糖凝胶电泳观察DNA梯形条带。结果:NOChR抑制HT-29细胞活性,呈浓度依赖性,其IC50值为1.78μM;NOChR具有诱导HT-29细胞凋亡作用;PPARγ特异性阻断剂GW 9662能部分拮抗NOChR诱导HT-29细胞凋亡。结论:NOChR诱导HT-29细胞凋亡作用与其活化PPARγ相关。     

Synthesis and PGE<Subscript>2</Subscript> inhibitory activity of vinylated and allylated chrysin analogues

Vinylated and allylated chrysin analogues were prepared as congeners of prenylated flavonoids and evaluated their anti-inflammatory activity. 8-Substituted chrysin analogues were prepared from 2'-hydroxy-3'-iodo-4',6'-dimethoxyacetophenone in 3 steps. 3-Allylated chrysin analogues were prepared from chrysin in 3 steps. Synthesized chrysin analogues (4, 5 and 8) showed moderate inhibitory activities of PGE2 production from LPS-induced RAW 264.7 cells.     

Disposition and metabolism of the flavonoid chrysin in normal volunteers

AIMS: To describe the oral disposition of the dietary flavonoid chrysin in healthy volunteers. METHODS: Oral 400 mg doses of chrysin were administered to seven subjects. Chrysin and metabolites were assayed in plasma, urine and faeces by h.p.l.c. RESULTS: Peak plasma chrysin concentrations were only 3-16 ng ml(-1) with AUCs of 5-193 ng ml(-1) h. Plasma chrysin sulphate concentrations were 30-fold higher (AUC 450-4220 ng ml(-1) h). In urine, chrysin and chrysin glucuronide accounted for 0.2-3.1 mg and 2-26 mg, respectively. Most of the dose appeared in faeces as chrysin. Parallel experiments in rats showed high bile concentrations of chrysin conjugates. CONCLUSIONS: These findings, together with previous data using Caco-2 cells, suggest that chrysin has low oral bioavailability, mainly due to extensive metabolism and efflux of metabolites back into the intestine for hydrolysis and faecal elimination.