DNA stability and lipid peroxidation in vitamin E–deficient rats in vivo and colon cells in vitro

Summary Background Fruit and vegetable consumption protects against cancer. This is attributed in part to antioxidants such as vitamin E combating oxidative DNA damage. Anthocyanins are found in significant concentrations in the human diet. However, it remains to be established whether they are bioactive in vivo. Aim To investigate the consequence both of vitamin E deficiency on oxidative damage to DNA and lipids and the cytoprotective effect of nutritionally relevant levels of cyanidin–3–glycoside both in vivo in rats and in vitro in human colonocytes. Methods Male Rowett Hooded Lister rats were fed a diet containing less than 0.5 mg/kg vitamin E or a vitamin E supplemented control diet containing 100 mg d–tocopherol acetate/kg. Half of the controls and vitamin E–deficient rats received cyanidin–3–glycoside (100 mg/kg). After 12 weeks endogenous DNA stability in rat lymphocytes (strand breaks and oxidised bases) and response to oxidative stress ex vivo (H₂O₂; 200 µM) was measured by single cell gel electrophoresis (SCGE). Tissue levels of 8–oxo–7,8–dihydro–2–deoxyguanosine (8–Oxo–dG) were measured by HPLC with EC detection. D–tocopherol and lipid peroxidation products (thiobarbituric acid reactive substances; TBARS) were measured by HPLC. Rat plasma pyruvate kinase and the production of reactive oxygen by phagocytes were detected spectrophotometrically and by flow cytometry respectively. Immortalised human colon epithelial cells (HCEC) were preincubated in vitro with the anthocyanins cyanidin and cyanidin–3–glycoside and the flavonol quercetin (all 50 µM) before exposure to H₂O₂ (200 µM). DNA damage was measured by SCGE as above. Results Plasma and liver d–tocopherol declined progressively over 12 weeks in rats made vitamin E deficient. Lipid peroxidation was increased significantly in plasma, liver and red cells. Reactive oxygen levels in phagocytes and plasma pyruvate kinase were increased. Vitamin E deficiency did not affect DNA stability in rat lymphocytes, liver or colon. Cyanidin–3–glycoside did not alter lipid peroxidation or DNA damage in rats. However, it was chemoprotective against DNA damage in human colonocytes.DNA strand breakage was decreased 38.8 ± 2.2 % after pretreatment with anthocyanin. Conclusion while it is accepted that vitamin E alters lipid oxidation in vivo, its role in maintaining DNA stability remains unclear. Moreover, whereas cyanidin–3–glycoside protects against oxidative DNA damage in vitro, at nutritionally relevant concentrations it is ineffective against oxidative stress in vivo.     

Tissue factor inhibitory sesquiterpene glycoside from<Emphasis Type="Italic">Eriobotrya japonica</Emphasis>

Tissue factor (TF, tissue thromboplastin) is a membrane bound glycoprotein, which accelerates the blood clotting, activating both the intrinsic and the extrinsic pathways to serve as a cofactor for activated factor VII (Vlla). The TF-factor Vlla complex (TF/Vlla) proteolytically activates factors IX and X, which leads to the generation of thrombin and fibrin clots. In order to isolate TF inhibitors, by means of a bioassay-directed chromatographic separation technique, from the leaves ofEriobotrya japonica Lindley (Rosaceae), a known sesquiterpene glycoside (2) and ferulic acid (3) were isolated as inhibitors that were evaluated using a single-clotting assay method for determining TF activity. Another sesquiterpene glycoside (1) was also isolated but was inactive in the assay system. Compound3 was yielded by alkaline hydrolysis of compound2. The structures of compounds1, 2, and3 were identified by means of spectral analysis as 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→6)]-β-D-glucopyranosyl nerolidol (1), 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyr-anosyl-(1→2)-[α-L-(4-trans-feruloyl)-rhamnopyranosyl-(1→6)]-β-D-glucopyranosyl nerolidol (2) and ferulic acid (3), respectively. Compounds2 and3 inhibited 50% of the TF activity at concentrations of 2 and 369 μM/TF units, respectively.     

Flavonol glycosides from the aerial parts of<Emphasis Type="Italic">Aceriphyllum rossii</Emphasis> and their antioxidant activities

The methanol extract obtained from the aerial parts ofAceriphyllum rossii (Saxifragaceae) was fractionated into ethyl acetate (EtOAc),n-BuOH and H₂O layers through solvent fractionation. Repeated silica gel column chromatography of EtOAc andn-BuOH layers afforded six flavonol glycosides. They were identified as kaempferol 3-O-β-D-glucopyranoside (astragalin,1), quercetin 3-O-β-D-glucopyranoside (isoquercitrin,2), kaempferol 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside (3), quercetin 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyrano-side (rutin,4), kaempferol 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (5) and quercetin 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (6) on the basis of several spectral data. The antioxidant activity of the six compounds was investigated using two free radicals such as the ABTS free radical and superoxide anion radical. Compound1 exhibited the highest antioxidant activity in the ABTS2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging method. 100 mg/L of compound1 was equivalent to 72.1±1.4 mg/L of vitamin C, and those of compounds3 and5 were equivalent to 62.7±0.5 mg/L and 54.3±1.3 mg/L of vitamin C, respectively. And in the superoxide anion radical scavenging method, compound5 exhibited the highest activity with an IC₅₀ value of 17.6 ± 0.3 μM. In addition, some physical and spectral data of the flavonoids were confirmed.     

Secoiridoid glucosides with free radical scavenging activity from the leaves of Syringa dilatata

Activity-guided fractionation of the EtOAc and MeOH extract of the leaves of Syringa dilatata NAKAI furnished one free radical scavenger, the secoiridoid glucoside oleuropein together with ligstroside and an iridoid glucoside, syringopicroside. Oleuropein interacted with the stable free radical, 1,1-diphenyl-2-picrylhydrazyl (DPPH), and showed an IC(50) value of 40.4 microM. L-Ascorbic acid as a positive control showed an IC(50) value of 50.3 microM.     

Urinary collagen metabolite excretion in amyotrophic lateral sclerosis

Collagen abnormalities of the spinal cord and the skin have been reported in patients with amyotrophic lateral sclerosis (ALS). The urinary concentrations of the hydroxylysine glycosides, i.e., glucosylgalactosyl hydroxylysine (glu-gal Hyl) and galactosyl hydroxylysine (gal Hyl), indicate the tissue origin of the collagen metabolites and the rate of the degradation of collagen. We measured the urinary levels of glu-gal Hyl and gal Hyl in 12 ALS patients, 10 diseased control subjects with other neurologic or muscular diseases (Control Group A), and 10 healthy control subjects (Control Group B). The urinary level of glu-gal Hyl in ALS patients was significantly lower than in the two control groups. In addition, a significant negative relationship between glu-gal Hyl urinary level and duration of illness was found in ALS patients. There was no marked difference in the urinary level of gal Hyl between ALS patients and the control groups. Our data suggest that the decreased urinary level of glu-gal Hyl may be useful in assessing the alteration in collagen metabolism in ALS and may have a relationship with the progression of ALS.     

不同色谱材料对黄酮苷分离研究

目的:考察3种反相柱色谱材料对黄酮苷类成分的分离.方法:采用RP-18、ODS和ODS-AQ 3种反相材料分离纯化黄酮苷类成分.结果:不同材料对黄酮苷类成分纯化效果不同.结论:ODS- AQ对饿蚂蝗黄酮苷纯化效果最好.     

炮制对何首乌中有效成分含量的影响

目的 :测定何首乌炮制前后蒽醌类和二苯乙烯苷的含量变化。方法 :大黄素对照品溶液与 0 .5 %Mg(AC) 2 显色 ,在5 10nm处以 0 .5 %Mg(AC) 2 为空白 ,测其吸收度 ;二苯乙烯苷对照溶液以 95 %乙醇稀释定容 ,在 310nm处测其吸收度。结果 :大黄素和二苯乙烯苷测定回归方程分别为Y =2 9.70X +7.95× 10 -3 ,r =0 .9993和Y =73.0 0X +0 .0 4 2 0 ,r =0 .99992。结论 :制何首乌中游离蒽醌的含量略高于生何首乌 ,而结合蒽醌的含量生何首乌则明显高于制何首乌 ;二苯乙烯苷的含量生何首乌要高于制何首乌     

胭脂花化学成分的研究Ⅱ

 目的研究报春花科报春花属植物胭脂花(Primula maximowiczii Regel)的化学成分。方法运用多种色谱方法,从胭脂花全草中分离得到9个化合物,通过理化性质和波谱数据鉴定化合物的结构。结果分离得到的化合物结构为:山柰酚3-O-β-D-葡萄糖基(1→2)-β-D-葡萄糖基(1→6)-β-D-葡萄糖苷(1),山柰酚3-O-(2″,6″-二-O-α-L-鼠李糖基)-β-D-半乳糖苷(2),槲皮素3-O-(2″,6″-二-O-α-L-鼠李糖基)-β-D-半乳糖苷(3),山柰酚3-O-β-D-半乳糖苷(4),槲皮素3-O-β-D-半乳糖苷(5),杨梅素3-O-β-D-半乳糖苷(6),山柰酚3-O-(2″,6″-二-O-α-L-鼠李糖基)-β-D-葡萄糖苷(7),槲皮素3-O-(2″,6″-二-O-α-L-鼠李糖基)-β-D-葡萄糖苷(8),樱草苷(9)。结论化合物1为新化合物,化合物2～8为首次从该属植物中分离得到,化合物9为首次从胭脂花中分离得到,同时本实验将化合物9的¹³C-NMR数据补充完整。     

肉苁蓉药材与盐生肉苁蓉培养细胞的主要成分对比研究

目的研究肉苁蓉药材与盐生肉苁蓉培养细胞的苯乙醇苷类成分差异。方法利用傅里叶变换红外光谱(FTIR)和HPLC技术进行成分对比分析。结果FTIR的分析结果表明，盐生肉苁蓉培养细胞的红外指纹谱图与肉苁蓉药材具有一定相似性；HPLC的分析结果表明，盐生肉苁蓉细胞中所含的主要成分和肉苁蓉药材相似，只是含量有差异。结论从整体上看，盐生肉苁蓉培养细胞中所含成分种类较多，且大部分含量较高，其中洋丁香酚苷(类叶升麻苷)和松果菊苷的含量大幅度高于肉苁蓉药材。     

不同炮制辅料对何首乌药效成分含量的影响

目的研究不同炮制辅料对何首乌药效成分含量的影响,为选择何首乌炮制辅料提供依据。方法分别采用5种不同炮制辅料制备制何首乌,高效液相色谱法按照2015年版中国药典中制何首乌成分含量测定方法分别测定制何首乌中二苯乙烯苷、游离蒽醌的含量,比较不同炮制辅料对何首乌中药效成分含量的影响。结果炮制辅料基本都会降低制何首乌中二苯乙烯苷和游离蒽醌的含量。不同炮制辅料制备的制何首乌中二苯乙烯苷含量从高到低依次为空白对照何首乌>米泔水制何首乌>生姜汁制何首乌>甘草汁制何首乌>熟地汁制何首乌>黑豆汁制何首乌,游离蒽醌含量从高到低依次为甘草汁制何首乌>空白对照何首乌>米泔水制何首乌>熟地汁制何首乌>黑豆汁制何首乌>生姜汁制何首乌。不同炮制辅料制备的何首乌中二苯乙烯苷含量均高于2015年版中国药典中的要求,而游离蒽醌含量均未达到2015年版中国药典中的要求。结论不同炮制辅料对制何首乌中药效成分含量的影响不同。