The flavonoids were extracted from alfalfa using ethanol assisted with ultrasonic extraction and purified by D101 macroporous resin column chromatography. The chemical composition and content of ethanol elution fractions (EEFs) were assessed by ultrahigh-performance liquid chromatography and hybrid quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS) and aluminum nitrate-sodium nitrite-sodium hydroxide colorimetric method. The in vitro antioxidant activity of two EEFs was conducted by scavenging DPPH free radical, and the main antioxidants of 75% EEFs were screened using DPPH-UHPLC. Moreover, the in vivo antioxidant activity of 75% EEFs and the growth performance of broilers were studied. The results showed that the content of 30% and 75% EEFs was 26.20% and 62.57%. Fifteen compounds were identified from 75% EEFs, and five of them were reported in alfalfa for the first time. The scavenging activity of 75% and 30% EEFs (200 μg/mL) against DPPH was 95.51% and 78.85%. The peak area of 5,3′,4′-trihydroxyflavone and hyperoside was decreased by 82.69% and 76.04%, which exhibited strong scavenging capacities. The total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) level of three treated groups against the normal control group (NC) fed with basal diet significantly increased by 3.89-24.49%, 0.53-7.39%, and 0.79-11.79%, respectively. While the malondialdehyde (MDA) decreased by 0.47-18.27%. Compared with the NC, the feed to gain ratio (F : G) of three treated groups was lowered by 2.98-16.53% and survival rate of broilers significantly increased. Consequently, 75% EEFs extracted from alfalfa exhibited powerful antioxidant activities and might be a potential feed additive to poultry and livestock.
试验以黄金茶为原料,通过超声波法提取黄金茶粗多糖.粗多糖经除脂肪、色素、蛋白等杂质,以0.3%NaCl为洗脱剂经DEAE-纤维素柱纯化后得到纯度为65.4%的黄金茶多糖.利用紫外光谱(UV)、红外光谱(FT-IR)、气相色谱(GC)、扫描电镜(SEM)初步对其结构进行探究;通过苯酚-硫酸法测定多糖含量,考马斯亮蓝法测定蛋白质含量.采用Sevage法对黄金茶多糖进行除蛋白工艺研究,在单因素试验基础上进行L9(33)正交试验,确定最佳脱蛋白工艺条件.结果表明:黄金茶多糖含有部分糖蛋白,含有-OH、-CH、-COOH、S=O、-C=O等官能团;黄金茶多糖由鼠李糖(Rha)、阿拉伯糖(Ara)、木糖(Xyl)、甘露糖(Man)、葡萄糖(Glu)、半乳糖(Gal)6种单糖组成,且6种单糖的摩尔比为0.069:0.148:0.056:0.051:0.566:0.102.黄金茶多糖的最佳除蛋白工艺条件为:Sevage试剂添加量为多糖溶液体积的1/3,振摇时间为20 min,氯仿与正丁醇体积比为4:1.
This study aimed to investigate the chemical composition and hepatoprotective effect of flavonoid fraction from Cyclocarya paliurus leaves (CPF) on the lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced acute liver failure (ALF). UPLC-MS analysis revealed that CPF mainly contained quercetin-3-O-glycoside, quercetin-3-O-rhamnoside, kaempferol-3-O-rhamnoside, quercetin and kaempferol. CPF treatment (100, 200, and 400 mg/kg) effectively prevented LPS/D-GalN-induced ALF by lessening the mortality; improving hepatic morphological structures and oxidative stress; decreasing the ALT and AST levels and inflammatory cytokines. Additionally, CPF alleviated hepatocyte apoptosis and mitochondrial dysfunction, which could be attributed to the activation of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase (HO)-1 pathway and the regulation of caspse-3, reactive oxygen species modulator 1 (Romo1) and B-cell lymphoma-extra large (Bcl-XL) expressions. Moreover, CPF dose-dependently inhibited the activation of NF-κB pathway and downregulated the expression of CD14 and TNF-R1. These results indicate that CPF could be developed as a promising functional food for the prevention of ALF.
The ethanol extracts of Chimonanthus nitens Oliv. leaves were prepared sequentially by ethanol gradient elution and tested for their α-glucosidase inhibitory. The fraction of 50% ethanol eluate (EE) exhibited the notable inhibition with IC50 of 0.376mg/mL. Also, 50% EE was chemically characterized by liquid chromatography–mass spectrometry (LC–MS) analysis. Eight compounds including rutin (1), hyperin (2), isoquercitrin (3), luteoloside (4), astragalin (6), quercetin (13), naringenin (14), kaempferol (15) were identified by compared with standard substances as well as proper luteolin-5-O-glucoside (5), kaempferol-7-O-rhamnoside (9), 5,7,8-trihydroxy-2-methoxyl-flavone-7-O-glucoside (10), kaempferol-7-O-acetyl-galactoside (11). The experiments of ultra-filtration combined with liquid chromatography–mass spectrometry (UF-LC–MS) guided quercetin and kaempferol as the key factors for 50% EE showing highly inhibitory activity on α-glucosidase. Quercetin and kaempferol inhibited yeast α-glucosidase in a mixed-type manner with IC50 of 66.8 and 109μg/mL, respectively. These results would provide theoretical underpinning for the C. nitens Oliv. leaves ethanol extracts used as nutraceutical health supplement in the management of type 2 diabetes.
The paper investigated the antihyperglycemic and antihyperlipidemic efficacy and antioxidant capacity of Chimonanthus nitens Oliv. leaf extract (COE) in combination of high-glucose-fat diet-fed and streptozotocin-induced diabetic model mice. Various physiological indexes in diabetic model mice were well improved especially by oral administration of high dose of COE; the results were listed as follows. Fast blood glucose (FBG) level and serum triglyceride (TC), total cholesterol (TG), low-density lipoprotein cholesterol (LDLC), and malondialdehyde (MDA) as well as MDA in liver were significantly reduced; fasting serum insulin (FINS) and insulin sensitivity index (ISI) were both increased; high-density lipoprotein cholesterol (HDLC) in serum was significantly increased; total antioxidant capacity (T-AOC), activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in serum and liver were apparently enhanced; liver coefficient (LC), liver transaminase, and alkaline phosphatase (ALP) were decreased. Furthermore, pancreas islets and liver in diabetic model mice showed some extend of improvement in morphology and function after 4 weeks of COE treatment. In consequence, COE was advantageous to regulate glycolipid metabolism and elevate antioxidant capacity in diabetic model mice. Thus, the present study will provide a scientific evidence for the use of COE in the management of diabetes and its related complications.
This study was designed to study the chemical composition of Cyclocarya paliurus polysaccharide and inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage. A new elution (0.3% NaCl aqueous solution) of Cyclocarya paliurus polysaccharide (CPP-3) was characterized by different methods such as fourier transform infrared spectra (FT-IR), UV-vis, gas chromatography-mass spectrometry (GC–MS), high performance gel chromatography (HPGLC) and scanning electron microscopy (SEM). Cell viability was measured by MTT test, phagocytosis assay was measured by Neutral red uptake assay, nitrite was measured by Griess assay, TNF-α and IL-1β analysis were measured by ELISA, PGE2 was measured by enzyme immunoassay system. The results showed that CPP-3 was comprised of two polysaccharides with average molecular weight (Mw) of 5.69×104Da and 4.94×103Da. CPP-3 contains six monosaccharides, of which are rhamnose (Rha), arabinose (Ara), xylose (Xyl), mannose (Man), glucose (Glu), galactose (Gal), the molar ratio of six monosaccharides is 0.060:0.109:0.053:0.128:0.293:0.357. CPP-3 increased the amount of NO released from mouse macrophage RAW264.7 and significantly increased the levels of TNF-α, IL-1β and PGE2 (P<0.01). CPP-3 suppressed LPS-stimulated RAW264.7 macrophage to release NO, TNF-α, IL-1β and PGE2 (P<0.01). CPP-3 and LPS accounted for synergistic effect on the release of NO and TNF-α, CPP-3 and LPS accounted for antagonistic effect on the release of IL-1β and PGE2.
The objective of this study was to analyse the structure of CPP-2, and to observe the pharmacological effects of CPP-2 on lipid metabolism and oxidative stress. CPP-2, eluted as two main fractions comprised of two polysaccharides with Mw of 307 and 3.7kDa, was mainly consisted of rhamnose, mannose, glucose and galactose in a molar ratio of 1.00:0.78:3.22:0.45. The results showed that treatment with CPP-2 could improve blood lipid levels (TC, TG, HDL-C and LDL-C), liver lipid levels (TC and TG) and antioxidant status (SOD, T-AOC, GSH-PX, MDA and LPO). In addition, the histopathological observations of mice livers and the GPT activities indicated that CPP-2 could attenuate liver cell injury. The present findings demonstrated that CPP-2 might be effective in lowering lipid and protecting against HFE-induced hyperlipidemia and non-alcoholic fatty liver.