染料木黄酮诱导前列腺癌DU-145细胞凋亡的研究

【摘要】〓目的〓探讨染料木黄酮诱导前列腺癌细胞系DU-145细胞凋亡的作用。方法〓在Caspase抑制剂Z-VAD存在或不存在的情况下,以染料木黄酮作用于前列腺癌DU-145细胞,采用MTT法检测细胞存活率、Annexin V-FITC/PI双染法流式细胞术检测细胞凋亡、PI染色法流式细胞术检测细胞周期、JC-1染色法流式细胞术测定细胞线粒体膜电位(△ψm),比色检测试剂盒测定caspase-8活性。结果〓染料木黄酮能明显降低前列腺癌DU-145细胞的存活率、诱导癌细胞凋亡、降低△ψm及提高caspase-8活性(全部P<0.05)。Z-VAD能够逆转Gen抗前列腺癌的效应。结论〓染料木黄酮使细胞阻滞于G0和G1,分别通过凋亡信号通路的外源性和内源性途径,诱导前列腺癌细胞凋亡。     

染料木素对去卵巢大鼠颌骨代谢的改善作用

目的 探讨染料木素对去势大鼠颌骨代谢及雌性生殖系统的作用。方法 切除成年雌性大鼠双侧卵巢，造成绝经后骨质疏松的动物模型。双能X线法检测颌骨、股骨骨密度，生化法检测血清碱性磷酸酶(alkaline phosphatase，ALP)、酸性磷酸酶(acid phosphatase，ACP)，放射免疫法检测骨代谢因子IL-1β、IL-6、转化生长因子α(tumor necrosis factor-α，TNF-α)、降钙素和骨钙素，探讨染料木素对去势大鼠颌骨代谢的作用；通过血清雌二醇含量和子宫重量的变化监测染料木素的雌激素样副作用。结果 去势大鼠骨密度显著减少；血清ALP、ACP和IL-1β、骨钙素显著高于非去势组。口服染料木素后，大鼠骨密度显著增加；血清ALP、ACP、骨钙素显著增高，其中ALP和骨钙素的增加显著优于雌激素组，IL-1β和TNF-α显著降低；大鼠子宫重量系数显著低于雌激素组。结论 染料木素能够通过抑制骨吸收和促进骨形成有效改善去势大鼠颌骨代谢，副作用弱于雌激素。染料木素在防治颌骨乃至全身骨质疏松方面具有开发前景。     

金雀异黄酮促进体外培养大鼠骨髓间充质干细胞成骨性分化的机制研究

目的 探讨金雀异黄酮促进体外培养原代大鼠骨髓间充质干细胞(rats bone marrow mesenchymal stem cell,rBMSC)成骨性分化的机制.方法 体外分离培养rBMSC传代3次后,采用碱性磷酸酶(ALP)活性筛选金雀异黄酮最佳浓度,采用终浓度为1×10-5 mol/L的金雀异黄酮对体外培养原代rBMSC进行干预;药物干预后48和72 h采用甲基噻唑基四唑(MTT)测定细胞增殖情况;在干预后3、6、9、12和15 d测定ALP活性、钙盐沉积量;干预后24、48和96 h检测OPG和RANKL mRNA表达水平.结果 金雀异黄酮组处理rBMSC 48和72 h后光密度值高于对照组(P＜0.05);干预rBMSC 12和15 d后金雀异黄酮组ALP活性高于对照组,干预6、9、12和15 d后钙盐沉积量高于对照组,干预24、48和72 h OPG mRNA水平高于对照组,干预72 h RANKL mRNA高于对照组,干预48 h OPG/RANKL高于对照组(P＜0.05,P＜0.01).结论 1×10-5 mol/L金雀异黄酮可促进体外培养rBMSC成骨性分化,其作用可能是通过OPG/RANKL途径实现的.     

染料木黄酮对成骨细胞增殖、分化和凋亡影响的实验研究

目的研究染料木黄酮对成骨细胞增殖、分化和凋亡的影响，以阐明其对骨形成的作用机制。方法酶消化和组织块相结合培养获得原代成骨细胞，并以成骨肉瘤细胞株UMR-106细胞作对照，加入不同浓度的染料木黄酮后，流式细胞仪和噻唑蓝比色法(MTT)检测成骨细胞细胞周期比例的改变以及凋亡情况；生化法检测细胞内碱性磷酸酶含量的改变。结果流式细胞分析和MT观测结果表明：染料木黄酮促进了原代成骨细胞由G0(G1)期向S期、G2期和M期的移行过渡，从而促进了原代成骨细胞的增殖。染料木黄酮对成骨肉瘤细胞株UMR-106细胞周期无影响，但可诱导UMR-106细胞凋亡。碱性磷酸酶检测结果表明：染料木黄酮可促进原代成骨细胞的分化。结论染料木黄酮可在一定程度上促进成骨细胞的增殖和分化，诱导成骨肉瘤细胞凋亡。     

Genistein upregulates placental corticotropin-releasing hormone expression in lipopolysaccharide-sensitized mice

Genistein is a phytoestrogen isolated from soyabeans. Although soy products are staple food of Asian, the potential effect of genistein on reproduction has not been fully addressed. Lipopolysaccharide (LPS) is an endotoxin found in the cell membrane of gram-negative bacteria. It may cause inflammation and other immune responses. Previous study has shown that LPS may induce pre-mature birth in rodents. In the present study, effect of genistein on LPS-induced preterm birth was investigated. Pregnant ICR mice were gavaged with genistein at 40, 200 and 400 mg/kg body weight/day during E13 to E16. LPS was injected i.p. on E16.5 and the animals were sacrificed at E17. Compared to the control group, an increased incidence of early delivery was observed in the pooled mice under LPS treatment. A rising trend of incidence was also demonstrated dose-dependently with genistein co-treatment. Real-time RT-PCR indicated that the placental crh expression was highly induced by the co-administration of 400 mg/kg genistein and LPS. By contrast, neither genistein nor LPS alone could alter the expression. Increased plasma CRH concentration was also seen in the co-treatment groups. In addition, the mRNA expression of placental CRH-binding protein and plasma progesterone concentration were reduced in these groups. These results indicated that genistein might exacerbate the undesirable effect of LPS on pregnant mice by altering hormonal regulations.     

Multi-targeted therapy of cancer by genistein

Soy isoflavones have been identified as dietary components having an important role in reducing the incidence of breast and prostate cancers in Asian countries. Genistein, the predominant isoflavone found in soy products, has been shown to inhibit the carcinogenesis in animal models. There is a growing body of experimental evidence showing that the inhibition of human cancer cell growth by genistein is mediated via the modulation of genes that are related to the control of cell cycle and apoptosis. It has been shown that genistein inhibits the activation of NF-kappaB and Akt signaling pathways, both of which are known to maintain a homeostatic balance between cell survival and apoptosis. Moreover, genistein antagonizes estrogen- and androgen-mediated signaling pathways in the processes of carcinogenesis. Furthermore, genistein has been found to have antioxidant properties, and shown to be a potent inhibitor of angiogenesis and metastasis. Taken together, both in vivo and in vitro studies have clearly shown that genistein, one of the major soy isoflavones is a promising agent for cancer chemoprevention and further suggest that it could be an adjunct to cancer therapy by virtue of its effects on reversing radioresistance and chemoresistance. In this review, we attempt to provide evidence for these preventive and therapeutic effects of genistein in a succinct manner highlighting comprehensive state-of-the-art knowledge regarding its multi-targeted biological and molecular effects in cancer cells.     

Genistein对APP695转染PC12细胞凋亡的保护作用

目的观察Genisteint（GST）对APP695转染PCI2细胞凋亡的保护作用。方法将培养的PC12细胞分为正常对照组、APP695转染组、GST干预组。采用流式细胞术检测PC12细胞凋亡率。结果APP695转染组与正常对照组比较,PC12细胞存活率明显降低,凋亡率明显升高（P〈0,01）。GST干预组与APP695转染组比较,细胞存活率显著升高,凋亡率显著降低,其中1μmol／LGST组与APP695转染纽此较,细胞凋亡率降低（P〈0．05）,5μmol／LGST组较APP695转染组细胞凋亡率则明显降低（P〈0．01）。结论GST对APP695基因转染引起的PC12细胞损伤有一定的保护作用。     

金雀异黄酮对糖尿病大鼠心肌损伤的保护作用

目的 观察金雀异黄酮（genistein, Gen）对糖尿病大鼠心肌损伤的保护作用,并探讨其机制。 方法 雄性SD大鼠随机分为正常（N）组、糖尿病（D）组、Gen 5 mg/kg治疗（L）组和Gen 25 mg/kg治疗（H）组,每组8只。采用链脲佐菌素55 mg/kg腹腔注射诱导1型糖尿病大鼠模型。造模成功4周后,L组和H组大鼠开始分别灌胃给予Gen溶液5 mg/kg和25 mg/kg。干预4周后,测定各组大鼠血流动力学指标和空腹血糖（FBG）;通过HE染色和透射电镜下分别观察大鼠心肌病理形态结构和超微结构改变;测定心肌组织肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）、白介素-6（IL-6）、丙二醛（MDA）、谷胱甘肽（GSH）含量和Caspase-3活性;RT-PCR检测心肌组织Bcl-2和BaxmRNA表达;Western blot检测心肌硫氧还蛋白（Trx）、Trx相互作用蛋白（TXNIP）和凋亡信号调节激酶1（ASK1）的蛋白表达。 结果 与N组比较,D组FBG、TNF-α、IL-1β、IL-6、MDA含量和Caspase-3活性增高（P＜0.01）,血流动力学指标和GSH含量下降（P＜0.01）;心肌病理形态结构和超微结构明显损伤,心肌Bcl-2 mRNA和Trx蛋白表达下降（P＜0.01）,BaxmRNA、TXNIP和ASK1蛋白表达增高（P＜0.01）。与D组比较,L和H组中,FBG差异无统计学意义,TNF-α、IL-1β、IL-6、MDA含量和Caspase-3活性下降（P＜0.05,P＜0.01）,血流动力学指标和GSH含量增高（P＜0.05,P＜0.01）;心肌病理形态结构和超微结构明显改善;心肌Bcl-2 mRNA和Trx蛋白表达增加（P＜0.05,P＜0.01）,Bax mRNA、TXNIP和ASK1蛋白表达下降（P＜0.05,P＜0.01）。 结论 金雀异黄酮对糖尿病大鼠心肌损伤具有保护作用,其机制可能与减轻心肌炎症反应,调节Trx系统表达,抑制氧化应激和细胞凋亡相关。     

金雀异黄素抑制大鼠膀胱肿瘤作用的实验研究

目的探讨金雀异黄素对N-甲基亚硝基脲(MNU)诱导的SD大鼠膀胱肿瘤的抑制作用。方法将60只大鼠随机分成3组,用MNU诱导成膀胱肿瘤,实验1组和实验2组分别经腹腔注射金雀异黄素0.5、1.0mg/kg,每两日1次;对照组用同样的方式注射生理盐水。利用统计方法和病理分级来分析金雀异黄素对MNU诱导的SD大鼠膀胱肿瘤的抑制作用。结果实验1组和实验2组与对照组比较,在体质量减少值、肿瘤个数、抑瘤率等方面,差异有统计学意义(P<0.05);实验1组和实验2组的癌变率相比,差异有统计学意义(P<0.05)。结论金雀异黄素对大鼠膀胱肿瘤有抑制作用,剂量越大,对肿瘤癌变抑制作用越强。