基于指纹图谱和UPLC-MS/MS定量测定对不同产地陈皮的质量评价研究

目的 采用HPLC指纹图谱与化学计量学相结合、UPLC-MS/MS测定多黄酮成分含量的方法,测定不同产地陈皮质量,为其鉴别和质量控制提供依据。方法 采用HPLC-DAD指纹图谱检测并结合“中药色谱指纹图谱相似度评价系统（2012A版）”建立6个产区,49个不同批次陈皮药材（S1～S49）指纹图谱,并进行相似度评价和共有峰确认,结合聚类分析（hierarchical clustering analysis,HCA）、主成分分析（principal component analysis,PCA）、正交偏最小二乘法-判别分析（orthogonal partial least-squares discrimination analysis,OPLS-DA）等化学计量学综合分析。采用UPLC-MS/MS法建立了8个黄酮类成分柚皮素、柚皮苷、木犀草苷、川陈皮素、橙皮素、橙皮苷、橘皮素和芸香柚皮苷的含量测定方法对60批7个产地陈皮药材进行评价。结果 建立了49批不同产地陈皮药材的指纹图谱,相似度为0.864～0.999,共标定了11个共有峰,HCA分析49批陈皮明显分为4类;PCA得到5个主成分的累积方差贡献率为92.748%;OPLS-DA表明8、7、10和11号峰可能是影响陈皮药材质量的差异标志物;含量测定结果表明不同产地陈皮中黄酮类含量存在显著差异,广东新会以川陈皮素和橘皮素为主要标志成分,四川荷花池以木犀草苷为差异性成分,江西樟树以柚皮素和柚皮苷为差异性成分,广西玉林则以橙皮素、橙皮苷和芸香柚皮苷为主,重庆云阳以川陈皮素和橙皮苷为主。结论 建立了稳定性强的不同产地陈皮HPLC指纹图谱和8个黄酮类UPLC-MS/MS定量测定方法,结合化学计量学可用于陈皮药材综合评价和质量控制。     

普洱茶籽的化学成分研究

普洱茶籽乙醇提出物的正丁醇萃取部分,经酸水解后,分离出5种成分,经光谱和化学分析,分别鉴定为二十八烷烃,柚皮素和3种新酯酰基皂甙元22-O-当归酸酯茶皂甙元B,22-O-当归酸酯茶皂甙元E和22-O-当归酸酯茶皂甙元A。     

Impact of Naringenin on Oxytetracycline-Mediated Oxidative Damage in Kidney of Rats

The aim of this study was to investigate the effect of naringenin on oxytetracycline-induced nephrotoxicity in rats. Oxytetracycline (200 mg/kg body weight, ip) was administered in 0.5ml of sterile physiological saline for 15 days, resulting in a significant increase in serum urea and creatinine and reduction in creatinine clearance. A significant increase in lipid peroxidation markers (TBARS and lipid hydroperoxide) and decrease in antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) and low molecular weight antioxidants (vitamin C, vitamin E, and reduced glutathione) levels were also observed in oxytetracycline-treated rats. The oral administration of naringenin (50 mg/kg body weight) attenuated the oxytetracycline-induced nephrotoxicity by significantly decreased levels of serum urea and creatinine with the significant normalization of creatinine clearance. Upon the administration of naringenin, the depleted renal antioxidant defense system (enzymatic and non-enzymatic antioxidants) was significantly increased in rats treated with oxytetracycline. These biochemical observations were supplemented by histopathological examination of kidney section. The present results suggest that the supplementation of naringenin might be helpful to alleviate the oxytetracycline-induced oxidative injury in kidney.     

核桃花化学成分的研究

目的 研究核桃Juglans regia花的化学成分。方法 利用正和反相硅胶、Sephadex LH-20及制备液相等柱色谱方法进行分离纯化,根据谱学数据及理化性质鉴定化合物结构,并采用MTT法检测化合物的体外细胞毒活性。结果 从核桃花95％乙醇水提取物中分离得到12个黄酮类化合物和7个生物碱类化合物,分别鉴定为金合欢素（1）、柚皮素（2）、5,7-二羟基-6,8,4''-三甲氧基黄酮（3）、木犀草素（4）、草质素（5）、白杨素（6）、异野樱素（7）、3,5-二羟基-4'',7-二甲氧基黄酮醇（8）、槲皮苷（9）、异槲皮苷（10）、山柰酚（11）、槲皮素（12）、2-（4-羟基苯乙基）异吲哚啉-1,3-二酮（13）、吲哚-3-甲酸乙酯（14）、1,2,3,4-四氢去甲哈尔满-1-酮（15）、3-醛基吲哚（16）、马齿苋酰胺E（17）、青兰素C（18）、N-苯甲酰-L-苯丙氨酸（19）。结论 除化合物4和9～12外,其余化合物均为首次从胡桃属中分离得到,其中化合物13为1个新的天然产物。化合物2、3和5具有抑制人结肠癌HCT-116细胞、人肝癌HepG2细胞、人胃癌BGC-823细胞、人肺支气管癌NCI-H1650细胞、人卵巢癌A2780细胞的增殖作用。     

Naringenin inhibited migration and invasion of glioblastoma cells through multiple mechanisms

Glioblastoma (GBM) is the most mortality brain cancer in the world. Due to high invasion and drug resistance cause the poor prognosis of GBM. Naringenin, an ingredient of citrus, exhibits many cellular functions such as antioxidant, anti‐inflammation, and anticancer. Naringenin inhibits the migration of bladder and lung cancer via modulation of MMP‐2 and/or MMP‐9 activities, Naringenin inhibits migration and trigger apoptosis in gastric cancer cells through downregulation of AKT pathway. However, the effects of naringenin in GBM still remain to be elucidated. In this study, we reveal the molecular mechanisms of naringenin in the inhibition of migration and invasion in GBM. No overt alternation of cell proliferation was found in of GBM 8901 cells treated with different concentration of naringenin. Slight decreased cell viability was found in GBM 8401 cell treated with 200 and 300 μM naringenin. Significant reduction of migration and invasion as assayed by Boyden chamber analysis was found in of GBM cells treated with 100, 200, and 300 μM naringenin. Zymography analysis also revealed that the activities of MMP‐2 and MMP‐9 of GBM cells were significantly inhibited in response to 100, 200, or 300 μM naringenin treatment. Proteins of MMP‐2 and MMP‐9 were downregulated in naringenin treated GBM cells. In addition, naringenin also attenuated the activities of ERK and p38. Naringenin decreased mesenchymal markers (snail and slug) expression as revealed by Western blot analysis. Taken together, our findings indicated that naringenin eliminated the migration and invasion of GBM cells through multiple mechanisms including inhibition of MMPs, ERK, and p38 activities and modulation of EMT markers. Our results also suggested that naringenin may be a potential agent to prevent metastasis of GBM.     

Identification of potential therapeutic target of naringenin in breast cancer stem cells inhibition by bioinformatics and in vitro studies

Cancer therapy is a strategic measure in inhibiting breast cancer stem cell (BCSC) pathways. Naringenin, a citrus flavonoid, was found to increase breast cancer cells’ sensitivity to chemotherapeutic agents. Bioinformatics study and 3D tumorsphere in vitro modeling in breast cancer (mammosphere) were used in this study, which aims to explore the potential therapeutic targets of naringenin (PTTNs) in inhibiting BCSCs. Bioinformatic analyses identified direct target proteins (DTPs), indirect target proteins (ITPs), naringenin-mediated proteins (NMPs), BCSC regulatory genes, and PTTNs. The PTTNs were further analyzed for gene ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, protein–protein interaction (PPI) networks, and hub protein selection. Mammospheres were cultured in serum-free media. The effects of naringenin were measured by MTT-based cytotoxicity, mammosphere forming potential (MFP), colony formation, scratch wound-healing assay, and flow cytometry-based cell cycle analyses and apoptosis assays. Gene expression analysis was performed using real-time quantitative polymerase chain reaction (q-RT PCR). Bioinformatics analysis revealed p53 and estrogen receptor alpha (ERα) as PTTNs, and KEGG pathway enrichment analysis revealed that TGF-ß and Wnt/ß-catenin pathways are regulated by PTTNs. Naringenin demonstrated cytotoxicity and inhibited mammosphere and colony formation, migration, and epithelial to mesenchymal transition in the mammosphere. The mRNA of tumor suppressors P53 and ERα were downregulated in the mammosphere, but were significantly upregulated upon naringenin treatment. By modulating the P53 and ERα mRNA, naringenin has the potential of inhibiting BCSCs. Further studies on the molecular mechanism and formulation of naringenin in BCSCs would be beneficial for its development as a BCSC-targeting drug.     

RP-HPLC法测定鬼箭羽中3种活性成分的含量

目的:建立同时测定鬼箭羽中降糖活性成分山柰酚、柚皮素及槲皮素含量测定的方法。方法:采用反相高效液相色谱法,以AlltimaC18(4.6mm×250mm,5μm)为色谱柱,乙腈-0.1%甲酸(38:62)为流动相,检测波长258nm,流速0.8mL/min,柱温30℃。结果:山柰酚、柚皮素及槲皮素的线性范围分别为0.02064~0.2580μg(r=0.9996),0.05592~0.699μg(r=0.9997),0.01656~0.207μg(r=0.9998);加样回收率分别为100.6%(RSD=2.5%),96.4%(RSD=1.1%),101.9%(RSD=1.7%)。结论:本法简便、准确、重现性好,适用于鬼箭羽中3种活性成分的同时测定,可用于鬼箭羽药材的质量控制。     

柚皮素和橙皮素对重组人肌醇磷脂3-激酶p110β催化亚基的影响

目的:研究柚皮素和橙皮素对重组人肌醇磷脂3-激酶(PI3-K)p110β催化亚基酶活性的影响。方法:通过基因工程技术的方法获得PI3-Kp110β催化亚基。用PtdIns(4,5)P2,[γ-32P]ATP与重组PI3-Kp110β催化亚基一起保温的方法测定PI3-K的活性;32P标记的磷脂用氯仿和甲醇抽提,加闪烁液于液闪计数仪计数。结果:柚皮素对重组人PI3-Kp110β催化亚基有抑制作用,IC50为10.57μmol/L;但橙皮素对重组人PI3-K没有抑制作用。结论:柚皮素是PI3-K的抑制剂。重组人PI3-Kp110β催化亚基可作为一种较为简便地筛选和开发有效的PI3-K抑制剂的分子靶点。     

Effect of domestic cooking on human bioavailability of naringenin,chlorogenic acid,lycopene and β-carotene in cherry tomatoes

Summary Background Epidemiological data showed that tomato and tomato product (sauce, paste) consumption is associated with a protective effect against the development of some chronic-degenerative diseases. Tomato antioxidant bioactive molecules such as carotenoids and polyphenols could be responsible, at least in part, for the healthy effect observed. The bioavailability of these compounds is an essential requirement to sustain their in vivo role. While it is well known that many factors can influence the bioaccessibility of carotenoids from the food matrix, there is little information about the factors affecting phenolic compounds bioaccessibility. Aim of the study This investigation was carried out to evaluate the effect of domestic cooking on the bioavailability in humans of antioxidant molecules after the administration of a test meal containing cherry tomatoes. Methods A cross-over design was conducted. Subjects (3 females and 2 males) consumed experimental meals containing fresh and cooked cherry tomatoes. Blood collection was performed at different time intervals (0, 2, 4, 6, 8 and 24 h). Results Carotenoid and phenol plasma concentrations were measured. Plasma levels of lycopene and -carotene were not significantly different with respect to the baseline after ingestion of both the test meals, while plasma concentrations of naringenin and chlorogenic acid increased significantly with respect to the baseline (P<0.05) after administration of cooked cherry tomatoes, but not after administration of fresh cherry tomatoes. Conclusions The present study indicated that domestically cooked tomatoes significantly increase naringenin and chlorogenic acid plasma levels. Considering that both naringenin and chlorogenic acid are widely studied for their potential healthy properties, evidence of their bioavailability and of the factors influencing their bioaccessibility is an important tool to sustain the possibility that these polyphenols play a biological role in human physiology.     

柚皮素对ARPE-19和HUVEC细胞的抗氧化作用

目的：研究柚皮素对人视网膜色素上皮细胞（ARPE-19）和人脐静脉内皮细胞（HUVEC）的抗氧化作用。 方法：采用MTT的方法检测ARPE-19和HUVEC细胞的生存率及增殖率。 结果：3，10mg／L柚皮素能显著增加ARPE-19细胞的增殖率达10．8％和11．4％。10mg／L柚皮素能提高ARPE-19细胞在缺氧，0．3mmol／LNaN，及200μmol／L，H2O2条件下的生存率分别为55．2％，69．2％及50．3％。1mg／L柚皮素能提高ARPE-19细胞在50μmol／L t-BHP和30mg／LNaIO3条件下的生存率达20．2％和30．4％。30mg／L柚皮素能够促进ARPE-19细胞在50μmol／L t-BHP条件下的增殖率达32．2％，而1mg／L柚皮素可以提高30，100，300mg／LNaIO3处理的ARPE-19细胞的增殖率达30．3％，10．3％及18．5％。3，10及30mg／L柚皮素抑制HUVEC的增殖率分别为23．9％，70．4％及77．9％。1，3mg／L柚皮素能提高HUVEC细胞在缺氧条件下的生存率达10．7％和13．1％，以及提高在300mg／LNaIO3条件下的生存率达41．2％和37．7％。3mg／L柚皮素能提高HUVEC细胞在200，400μmol／L H2O2条件下的生存率达20．1％和21．5％． 结论：柚皮素能够促进ARPE-19细胞的增殖率，抑制HU-VEC生长，同时对这两种细胞均有抗氧化作用。因此，柚臾素是治疗老年黄斑变性的很有前景的候选药物。