壮药两粤黄檀化学成分研究(I)

目的 研究壮药两粤黄檀Dalbergia benthami的化学成分。方法 通过硅胶、凝胶、HPLC等柱色谱及重结晶方法进行分离和纯化,根据理化性质和波谱数据对化合物结构进行鉴定。结果 从两粤黄檀乙醇提取物中分离得到14个化合物,分别鉴定为4''-羟基-6-羟甲基-5,7-二甲氧基异黄酮（1）、β-谷甾醇（2）、胡萝卜苷（3）、豆甾醇（4）、大鱼藤树酸（5）、robustin（6）、4-hydroxy-3-（3''-hydroxy-4''-methoxyphenyl）-5-methoxy-2",2"-dimethylpyrano-（5",6"：6,7）-coumarin（7）、大鱼藤树素甲醚（8）、大鱼藤树酸甲酯（9）、4''-羟基-5,7-二甲氧基-6-（3-甲基-2-丁烯基）-异黄酮（10）、芒柄花黄素（11）、伪赝靛苷元（12）、毛蕊异黄酮（13）、豆甾醇-3-O-β-D-葡萄糖苷（14）。结论 化合物1为新化合物,命名为两粤黄檀酮;所有化合物均为首次从该属植物中分离得到。     

Daidzein affects proliferation and apoptosis in non-small cell lung cancer cells:role of p53 signaling pathway北大核心CSCD

Aim To investigate the effects of daidzein（DD） on the proliferation and apoptosis of non-small cell lung cancer cells,with a focus on the possible role of the p53 signaling pathway in this regard. Methods CCK-8 method and flow cytometry were used to detect the effects of soy isoflavone crude extract and DD on the viability and apoptosis of HELF and H1299 cells. Gene microarray was used to detect the changes in gene expression after treatment of H1299 cells with DD. GSEA and differential analysis were used to screen the major pathways and key genes. RT-qPCR and Western blot were performed to verify the differences in mRNA and protein expression of key genes（p53 and CASP9） in the major pathways. After p53 inhibitor Pifithrin-α inhibited the expression of p53,the effect of DD on p53 mRNA and protein expression levels was examined,and the proliferative effect on H1299 cells was observed. Results Soy isoflavone crude extract and DD promoted proliferation and inhibited apoptosis of normal lung cells and inhibited proliferation and promoted apoptosis of lung cancer cells. p53 signaling pathway was significantly enriched in the DD-treated group（NES=1.78,P=0.000）,and the expressions of p53 and CASP9 genes were found to be significantly up-regulated in the treated group. Compared with the control group,mRNA expression of CASP9 and p53 significantly increased in both HELF and H1299 cells treated with DD（P<0.05）,and p53 protein expression also increased in HELF cells（P<0.05）. After inhibition of p53 expression,DD significantly increased the mRNA expression of p53 in H1299 and HELF cells（P<0.05） and also markedly increased the expression of p53 protein in H1299 cells（P<0.05）,and it was observed that DD inhibited the proliferation of lung cancer cells. Conclusions DD inhibits the proliferation and promotes the apoptosis of lung cancer H1299 cells,and the mechanism mainly involves the p53 signaling pathway. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

大豆异黄酮对去势大鼠骨质疏松性骨折愈合的影响

目的研究大豆异黄酮在大鼠骨质疏松性骨折愈合过程中的干预效果。方法将50只6月龄雌性SD大鼠随机分成5组:正常对照组、骨折组、骨质疏松组、骨质疏松性骨折组、骨质疏松性骨折治疗组,每组10只。正常对照组、骨折组只进行手术入路而不切除卵巢。骨质疏松组、骨质疏松性骨折组、骨质疏松性骨折治疗组摘除双侧卵巢制备骨质疏松模型。8周后,骨折组、骨质疏松性骨折组、骨质疏松性骨折治疗组制备右侧股骨干中段横行骨折模型。骨质疏松性骨折治疗组每日灌胃大豆异黄酮50 mg/kg(剂量1.5 ml/kg),骨折组、骨质疏松性骨折组灌胃等量生理盐水,连续8周。结果去卵巢后8周,骨质疏松组的骨密度显著低于正常对照组(P0.05),说明骨质疏松模型制作成功。灌胃8周后,骨质疏松性骨折组骨痂组织骨小梁数量及致密度、骨密度、血管内皮细胞生长因子的表达均显著低于骨折组和骨质疏松性骨折治疗组(P0.05)。结论大豆异黄酮在大鼠骨质疏松性骨折愈合过程中具有积极促进作用。     

大豆异黄酮抑制大鼠前列腺增生及其作用机制

目的观察大豆异黄酮(SI)对良性前列腺增生(BPH)大鼠体内性激素、生长因子及细胞凋亡相关基因的影响,探讨SI对BPH的防治及作用机制。方法选择SPF级健康成年雄性SD大鼠100只,随机分为正常对照(NC)组、BPH模型组、低剂量[6 mg/(kg·d)]SI组、中剂量[12 mg/(kg·d)]SI组及高剂量[24 mg/(kg·d)]SI组,每组各20只。灌胃给药4周后麻醉处理大鼠,腹主动脉取血,制备血清;完整摘取大鼠前列腺组织。称重测定前列腺组织湿质量,计算前列腺指数(PI);酶联免疫吸附法(ELISA)测定大鼠血清雌二醇(E2)、睾酮(T)水平;免疫质印迹法(Western blotting)检测前列腺组织Fas、Fas L、Bax、Bcl-2、表皮生长因子(EGF)及其受体EGFR的表达。结果与NC组比,BPH组前列腺组织湿质量及PI均较明显增加(P<0.05);血清E2及T水平均显著升高(P<0.05);前列腺组织中Fas L、Bcl-2、EGF及EGFR表达水平均显著升高(P<0.05),而Fas和Bax表达水平均显著降低(P<0.05)。与BPH组比,中、高剂量SI组前列腺组织湿质量及PI均明显减少(P<0.05);血清E2及T水平均显著下降(P<0.05);前列腺组织中Fas L、Bcl-2、EGF及EGFR表达水平均显著降低(P<0.05),而Fas和Bax表达水平显著升高(P<0.05);其中,中剂量SI组各指标变化较低、高剂量组更显著(P<0.05)。结论 SI对BPH具有较好的抑制作用,其中中剂量SI作用效果更佳,其作用机制可能与调节机体性激素水平,下调生长因子及其受体表达以及调控细胞凋亡基因表达有关。     

大别山野葛根异黄酮超声辅助提取工艺的响应面优化与还原力测定

 目的 确定大别山野葛根异黄酮的超声辅助最佳提取工艺与还原力。方法 在单因素实验的基础上,进行4因素5水平的Box-Behnken中心组合实验设计,利用响应面分析优化工艺条件,并测定优化工艺条件下葛根异黄酮提取液的还原力。结果 大别山野葛根异黄酮300 W功率的超声辅助提取最佳工艺条件为乙醇75%、液料比35 mL·g^-1、处理时间35 min、处理温度50 ℃,此工艺条件下葛根异黄酮得率为24.18%,相对误差仅3.02%;葛根异黄酮还原力的EC₅₀值为0.135 mg·mL^-1。结论 响应面法适用于对葛根异黄酮超声辅助提取得率进行回归分析和参数优化。     

大豆异黄酮对培养心肌细胞钙电流和动作电位的影响

目的 观察大豆异黄酮（SI）对大鼠心肌细胞动作电位的影响和培养心室肌细胞钙电流的作用。方法 采用悬浮玻璃微电极法和全细胞膜片钳方法。结果 SI能剂量依赖性地降低动作电位幅值（APA）、超射（OS）、最大除极速率（Vmax）、缩短复极50％及90％水平的动作电位时程（APD50、APD90）；SI 0.1pg/ml对L-型钙电流可见不同程度的抑制作用。结论 SI可使心肌细胞动作电位除极参数降低，动作电位时程缩短，对钙电流的抑制作用可减轻心肌细胞钙超载，进而保护心肌。     

Isoflavone supplementation reduces DNA oxidative damage and increases O-β-N-acetyl-d-glucosaminidase activity in healthy women

Phenolic compounds are believed to boost the human antioxidant defense system and health; therefore, the aim of this research was to investigate the hypothesis that soy isoflavones (IFs) provide antioxidant protection in healthy women by evaluating DNA resistance to oxidative damage and O-β-N-acetyl-d-glucosaminidase (OGA) activity. An IF supplement (80 mg/d) was given to 9 postmenopausal women and 13 young women for 6 months and then stopped up to the 14th month. The women were allowed to consume their normal diet. Blood samples were collected at the beginning of the study after 2, 4, and 6 months and then at the 8th and 14th months. Plasma concentrations of genistein and daidzein, total antioxidant capacity, plasma vitamin status, markers of oxidative stress (red blood cell membrane fluidity, activity of the red blood cell cytosolic enzyme OGA and lymphocyte DNA susceptibility to oxidative stress), and serum lipid profile were analyzed. Analysis of variance for repeated measures was used for statistical analysis. Plasma concentrations of IFs rose significantly during the supplementation period, and plasma total antioxidant capacity increased in young women; membrane fluidity and OGA activity increased, and DNA oxidative damage decreased (P < .05) at 4 months, then returned to the basal level. There was a significant inverse correlation between DNA damage and plasma IF concentrations (P < .01). The results indicated a positive effect of IF supplementation on oxidative stress in women, thus suggesting that the healthful action ascribed to soy consumption may be partially related to the antioxidant potential of IFs.     

Puerarin inhibits angiotensin II-induced cardiac hypertrophy via the redox-sensitive ERK1/2, p38 and NF-κB pathways

Aim： To investigate the effects of puerarin （Pue）, an isoflavone derived from Kudzu roots, on angiotensin II （Ang II）-induced hypertrophy of cardiomyocytes in vivo and in vitro.
Methods： C57BL/6J mice were infused with Ang II and treated with Pue （100 mg·kg-1·d-1, po） for 15 d. After the treatment, systolic blood pressure （SBP） and left ventricular wall thickness were assessed. The ratios of heart weight to body weight （HW/BW） and left ventricular weight to body weight （LVW/BW） were determined, and heart morphometry was assessed. Expression of fetal-type genes （ANP, BNP and β-MHC） in left ventricles was measured using semi-quantitative RT-PCR. Mouse primary cardiomyocytes were treated with Pue （50, 100, 200 μmol/L）, then exposed to Ang II （1 μmol/L）. ROS level was examined with flow cytometry, the binding activity of NF-κB was determined using EMSA. Western blot was used to measure the levels of ERK1/2, p38 and NF-κB pathway proteins. [3H]leucine incorporation was used to measure the rate of protein synthesis.
Results： Oral administration of Pue significantly suppressed Ang II-induced increases in the myocyte surface area, HW/BW, LVW/BW, SBP and left ventricular wall thickness. Furthermore, Pue significantly suppressed Ang II-induced increases in ANP, BNP and β-MHC expression in the left ventricles in vivo. Treatment of cardiomyocytes with Pue （50–500 μmol/L） did not affect the viability of cardiomyocytes in vitro. Pretreatment of cardiomyocytes with Pue dose-dependently inhibited Ang II-induced increases in ROS production, NF-κB binding activity, protein synthesis and cell breadth. Furthermore, pretreatment with Pue significantly suppressed Ang II-induced activation of ERK1/2, p38 and the NF-κB pathway proteins and the expression of ANP and β-MHC in cardiomyocytes. The positive drug valsartan exerted similar effects on Ang II-induced cardiac hypertrophy in vivo and in vitro.
Conclusion： Pue attenuates Ang II-induced cardiac hypertrophy by inhibiting activation of the redox-sensitive ERK1/2, p38 and the NF-κB pathways.     

大豆异黄酮对慢性肾损害大鼠肾功能及脂代谢的影响

目的观察不同剂量大豆异黄酮（SIF）对慢性肾损害大鼠肾功能及脂质代谢的影响。方法采用腺嘌呤灌喂方法建立肾损害大鼠模型，按体质量大小编号随机分为3组，每组9只。分别为肘照组，予同等量花生油灌喂；SIF小剂量组，予SIF6mg/（kg·d）；大剂量组，予SIF12mg/（kg·d）。观察第0天、第21天、第42天血浆甘油三酯、总胆固醇、高密度脂蛋白、低密度脂蛋白、血浆尿素、血清肌酐水平以及尿蛋白排泄量。结果SIF治疗组血浆尿素、血清肌酐水平下降及尿蛋白排泄量减少，血浆甘油三酯、总胆固醇、低密度脂蛋白下降，高密度脂蛋白水平升高，上述各项指标与对照组相比，差异均有统计学意义（P〈0．05），SIF不同剂量组之间比较差异无统计学意义（P〉0．05）。结论低剂量SIF就可起到改善肾损害大鼠的脂代谢紊乱，减少尿蛋白排泄和稳定肾功能的作用。     

新型金雀异黄素衍生物5-羟基-4'-硝基-7-取代酰氧基异黄酮的合成及抗肿瘤活性研究

目的:对金雀异黄素进行结构改造,设计合成7-取代酰氧基-5-羟基-4'-硝基金雀异黄素衍生物,并进行抗肿瘤活性测试.方法:以氯苄为起始原料,经取代、硝化、Friedel-Crafts反应和环合,再与各种酰氯反应得到8个目标化合物.结果:合成的目标化合物经元素分析和1HNMR确证其结构;其中化合物5B(5-羟基-4'-硝-7-丙酰氧基异黄酮)体内外对人乳腺癌细胞株MDA-MB-435均具有较强的抑制活性,IC50为0.017 mmol/L.结论: 金雀异黄素的4'引入硝基,同时7位丙酰化产物具有良好的抗肿瘤活性.