紫铆花素抑制脂多糖诱导的骨髓源性巨噬细胞炎症反应的研究

目的：研究紫铆花素（butein）对脂多糖（LPS）诱导小鼠骨髓源性巨噬细胞炎症反应的抑制效应，并探讨JAK2-STAT3通路在其中的作用。方法：分离C57BL/6小鼠骨髓，用40 ng/mL的巨噬细胞集落刺激因子刺激7 d，诱导为骨髓源性巨噬细胞（BMDM）。采用500 ng/mL的LPS刺激BMDM 12 h建立炎症模型，butein干预组采用5、10、20 μmol/L butein与LPS共处理，butein单独处理组为20 μmol/L的butein处理12 h，并设立空白对照组。ELISA法检测BMDM培养液中TNF-α、IL-6和NO的水平；流式细胞术检测细胞内ROS水平；Western blot法检测细胞内iNOS、p-JAK2、JAK2、p-STAT3和STAT3蛋白表达水平，并分析P-JAK2/JAK2和P-STAT3/STAT3蛋白表达水平的比值变化。结果：ELISA实验结果显示，LPS刺激BMDM后，培养液中的TNF-α、IL-6和NO含量显著升高，而5、10、20 μmol/L的butein干预可抑制上述促炎因子的分泌，且呈现剂量-效应关系。流式细胞术检测结果显示，butein抑制了LPS活化BMDM的ROS水平升高。Western blot检测结果表明，LPS刺激后BMDM内iNOS蛋白表达水平升高，JAK2和STAT3蛋白表达水平虽未明显变化，但磷酸化水平显著增加。而butein干预可有效抑制JAK2和STAT3蛋白磷酸化。结论：Butein可抑制LPS诱导的巨噬细胞炎症反应，JAK2-STAT3信号通路可能参与调控这一效应，提示butein是一种炎症相关疾病的候选药物。     

皂角刺中黄酮类成分及其抗肿瘤活性研究

目的研究皂角刺Gleditsiae Spina中黄酮类化学成分及其抗肿瘤活性。方法采用硅胶柱色谱、Sephadex LH-20柱色谱及半制备HPLC等方法对皂角刺中黄酮类化学成分进行分离纯化,并根据理化性质和波谱学数据分析鉴定化合物结构;采用SRB法测定化合物7~16对人乳腺癌细胞MCF-7的细胞毒活性。结果从皂角刺醋酸乙酯部位中分离得到16个黄酮类化合物,分别鉴定为tricin(1)、甘草素(2)、7,4′-二羟基-5,3′-二甲氧基黄酮醇(3)、鹰嘴豆醇(4)、7,3′,5′-三羟基二氢黄酮(5)、7,4′-二羟基黄酮醇(6)、双氢山柰素(7)、紫铆查耳酮(8)、(2S)-5,7,3′,5′-四羟基二氢黄酮(9)、7,3′,5′-三羟基-5-甲氧基黄酮醇(10)、槲皮素(11)、黄颜木素(12)、fisetin(13)、leucorobinetinidin(14)、thevetiaflavon(15)、异牡荆素(16);其中化合物8对MCF-7细胞的IC50值为28.53μmol/L。结论化合物1~6、8~10、14、15为首次从该属植物中分离得到,化合物8对MCF-7细胞具有明显细胞毒活性。     

Butein, a tetrahydroxychalcone, suppresses cancer-induced osteoclastogenesis through inhibition of receptor activator of nuclear factor-kappaB ligand signaling

Osteoclastogenesis is associated with aging and various age-related inflammatory chronic diseases, including cancer. Receptor activator of nuclear factor-kappaB (NF-κB) ligand (RANKL), a member of the tumor necrosis factor superfamily, has been implicated as a major mediator of bone resorption, suggesting that agents that can suppress RANKL signaling might inhibit osteoclastogenesis, a process closely linked to bone resorption. We therefore investigated whether butein, a tetrahydroxychalcone, could inhibit RANKL signaling and suppress osteoclastogenesis induced by RANKL or tumor cells. We found that human multiple myeloma cells (MM.1S and U266), breast tumor cells (MDA-MB-231) and prostate tumor cells (PC-3) induced differentiation of macrophages to osteoclasts, as indicated by tartrate-resistant acid phosphatase (TRAP)-positive cells, and that butein suppressed this process. The chalcone also suppressed the expression of RANKL by the tumor cells. We further found that butein suppressed RANKL-induced NF-κB activation and that this suppression correlated with the inhibition of IκBα kinase and suppression of phosphorylation and degradation of IκBα, an inhibitor of NF-κB. Finally, butein also suppressed the RANKL-induced differentiation of macrophages to osteoclasts in a dose-dependent and time-dependent manner. Collectively, our results indicate that butein suppresses the osteoclastogenesis induced by tumor cells and by RANKL, by suppression of the NF-κB activation pathway.     

Cyclooxygenase inhibitory flavonoids from the stem bark of Semecarpus anacardium Linn

The ethyl acetate extract of stem bark of Semecarpus anacardium showing in vivo anti-in fl ammatory activity in carrageenan induced rat paw edema assay was investigated in order to identify its active compounds. Chemical investigation of the ethyl acetate extract of S.anacardium afforded 3,4,2',4'-tetrahydroxychalcone (butein) and 7,3',4'-trihydroxy fl avone. Evaluation of COX-1 inhibitory activity of 3,4,2',4'-tetrahydroxychalcone and 7,3',4'-trihydroxy fl avone provided the IC(50) values of 28.4 and 36.7 micro M respectively. Further investigation of these compounds for COX-2 inhibitory activity revealed moderate potency towards this enzyme.     

Betulin, betulinic acid and butein are inhibitors of acetaldehyde-induced activation of liver stellate cells

Liver fibrosis has been reported to be inhibited in vivo by oleanolic and ursolic acids; however, the activity of other triterpenes like betulin and betulinic acid has not been examined. Butein has also been reported to prevent and partly reverse liver fibrosis in vivo, although its mechanism of action is poorly understood. Therefore, the aim of this study was to determine the antifibrotic potential of butein, betulin, and betulinic acid and examine their mechanisms of action in vitro. This study was conducted in rat stellate cells (HSCs) that were treated with acetaldehyde, which is the most reactive product of ethanol metabolism. Butein, betulin, and betulinic acid were preincubated with rat HSCs at non-toxic concentrations. Treatment effects were measured in regard to acetaldehyde-induced toxicity and cell migration, and several markers of HSC activation were evaluated, including smooth muscle α-actin (α-SMA) and procollagen I expression. In addition, changes in the release of reactive oxygen species (ROS) and cytokines such as tumor necrosis factor-α (TNF-α) and tumor growth factor-β1 (TGF-β1) and changes in the production of metalloproteinase-2 (MMP-2) and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) were determined. In vitro, HSCs were protected against acetaldehyde-induced toxicity by betulin but not by betulinic acid and butein. However, butein, betulin, and betulinic acid inhibited the production of ROS by HSCs treated with acetaldehyde and inhibited their migration. Butein also inhibited acetaldehyde-induced TGF-β1 production. Butein, betulin, and betulinic acid down-regulated acetaldehyde-induced production of TIMP-1 and TIMP-2. Betulin decreased the acetaldehyde-induced activity of MMP-2, but butein and betulinic acid did not. The results indicated that butein, betulin, and betulinic acid inhibited the acetaldehyde-induced activation of HSCs. Each drug functioned in a different manner, whereby some were acting as either antioxidants or inhibitors of TIMPs expression and butein additionally acted as an inhibitor of TGF-β production.     

An Investigation on the Therapeutic Potential of Butein,A Tretrahydroxychalcone Against Human Oral Squamous Cell Carcinoma

Background: Oral squamous cell carcinoma (OSCC) is one of the most predominant cancers in India. With advances in the field of oncology, a number of therapies have emerged; however, they are minimally effective. Consequently, there is a need to develop safe and effective regimens for the treatment of OSCC. Butein, a tetrahydroxychalcone has been found to exhibit potent antioxidant, anti-inflammatory, and also anti-tumor effects against several cancer types. However, its effect on OSCC is not studied yet. Methods: The effect of butein on the viability, apoptosis, migration and invasion of OSCC cells was evaluated using MTT, colony formation, PI/FACS, live and dead, scratch wound healing, and matrigel invasion assays. Further Western blot analysis was done to evaluate the expression of different proteins involved in the regulation of cancer hallmarks. Results: This is the first report exemplifying the anti-cancer effect of butein against OSCC. Our results showed that butein exhibited potent anti-proliferative, cytotoxic, anti-migratory, and anti-invasive effects in OSCC cells. It suppressed the expression of NF-κB and NF-κB-regulated gene products such as COX-2, survivin and MMP-9 which are involved in the regulation of different processes like proliferation, survival, invasion, and metastasis of OSCC cells. Conclusion Collectively, these results suggest that butein has immense potential in the management of OSCC. Nonetheless, in vivo validation is critical before moving to clinical trials.     

紫铆花素抑制脂多糖诱导的骨髓源性巨噬细胞炎症反应的研究

目的:研究紫铆花素(butein)对脂多糖(LPS)诱导小鼠骨髓源性巨噬细胞炎症反应的抑制效应,并探讨JAK2-STAT3通路在其中的作用。方法:分离C57BL/6小鼠骨髓,用40 ng/m L的巨噬细胞集落刺激因子刺激7 d,诱导为骨髓源性巨噬细胞(BMDM)。采用500 ng/m L的LPS刺激BMDM 12 h建立炎症模型,butein干预组采用5、10、20μmol/L butein与LPS共处理,butein单独处理组为20μmol/L的butein处理12 h,并设立空白对照组。ELISA法检测BMDM培养液中TNF-α、IL-6和NO的水平;流式细胞术检测细胞内ROS水平;Western blot法检测细胞内i NOS、p-JAK2、JAK2、p-STAT3和STAT3蛋白表达水平,并分析P-JAK2/JAK2和P-STAT3/STAT3蛋白表达水平的比值变化。结果:ELISA实验结果显示,LPS刺激BMDM后,培养液中的TNF-α、IL-6和NO含量显著升高,而5、10、20μmol/L的butein干预可抑制上述促炎因子的分泌,且呈现剂量-效应关系。流式细胞术检测结果显示,butein抑制了LPS活化BMDM的ROS水平升高。Western blot检测结果表明,LPS刺激后BMDM内i NOS蛋白表达水平升高,JAK2和STAT3蛋白表达水平虽未明显变化,但磷酸化水平显著增加。而butein干预可有效抑制JAK2和STAT3蛋白磷酸化。结论:Butein可抑制LPS诱导的巨噬细胞炎症反应,JAK2-STAT3信号通路可能参与调控这一效应,提示butein是一种炎症相关疾病的候选药物。     

肉豆蔻的化学成分研究

目的 对生品肉豆蔻Myristicae Semen的化学成分进行研究.方法 采用硅胶柱色谱、Sephadex LH-20柱色谱和制备型HPLC等方法进行分离纯化,结合波谱数据分析鉴定化合物结构.结果 从肉豆蔻75%醇提物的醋酸乙酯萃取物中分离得到18个化合物,分别鉴定为香草酸(1)、紫铆因(2)、(2R)-3-(3', 4', 5'-三甲氧基苯基)-1, 2-丙二醇(3)、硫磺菊素(4)、3-甲氧基-4, 5-亚甲二氧基肉桂酸(5)、7, 3', 4'-三羟基黄酮(6)、7-羟基色原酮(7)、verrucosin(8)、(+)-赤-(7S, 8R)-Δ^8'-7-羟基-3, 4, 3', 5'-四甲氧基-8-氧代-4'-新木脂素(9)、(-)-赤-(7R, 8S)-Δ^8'-7-乙酰基-3, 4, 3', 5'-四甲氧基-8-氧代-4'-新木脂素(10)、nectandrin B(11)、(-)-(7S, 7'R, 8S, 8'R)-4, 4'-二羟基-3, 5, 3'-三甲氧基-7, 7'-环氧木脂素(12)、fragransin B₃(13)、fragransin B₁(14)、(-)-enantiomer(15)、(-)-赤-(7R, 8S)-Δ^8'-7-羟基-3, 4, 5, 3', 5'-五甲氧基-8-氧代-4'-新木脂素(16)、(+)-赤-(7S, 8R)-Δ^8'-7, 4-二羟基-3, 5, 3', 5'-四甲氧基-8-氧代-4'-新木脂素(17)、(+)-5-甲氧基脱氢二异丁香酚(18).结论 化合物2、4～7为从肉豆蔻属植物中首次分离得到,化合物1为从该植物中首次分离得到.