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Objective: To investigate the antioxidant efficacy of oral orientin on 900 MHz radiofrequency-electromagnetic radiation-induced oxidative stress in mice. Methods: The mice were randomly allotted into 5 groups consisting of 7 mice each. The deionised water and radiofrequency electromagnetic radiations (RF-EMR) groups were administered with deionised water while orientin was administered to the RF-EMR + low dose of orientin group (10 mg/kg), RF-EMR + high dose of orientin group (20 mg/kg) and high dose of orientin group (20 mg/kg). All the groups except deionised water and high dose groups were exposed to 900 MHz radiofrequency-electromagnetic radiation for 28 consecutive days (1 h/day). Learning and memory was assessed via the step-down inhibitory avoidance task. Activities of lipid prexidation and antioxidant enzymes were measured using kits. Results: Radiofrequency electromagnetic radiation caused impairment in learning and memory and reduced activities of brain antioxidant enzymes, increased lipoperoxidation and corticosterone concentration as well as histopathological aberrations in the hippocampal tissues. Conversely, orientin alleviated learning and memory deficit, improved the activities of endogenous antioxidant enzymes and mitigated brain lipoperoxidation and neuronal degeneration in mice exposed to radiofrequency electromagnetic radiation. Conclusions: Orientin alleviates learning and memory impairment due to radiofrequency electromagnetic radiation in mice by improving antioxidant defence mechanism and may be considered as a promising therapeutic agent for improving the antioxidant system of people living in radiofrequency electromagnetic radiation-prone environment.
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Objective: To investigate the antioxidant efficacy of oral orientin on 900 MHz radiofrequencyelectromagnetic radiation-induced oxidative stress in mice. Methods: The mice were randomly allotted into 5 groups consisting of 7 mice each. The deionised water and radiofrequency electromagnetic radiations (RF-EMR) groups were administered with deionised water while orientin was administered to the RF-EMR + low dose of orientin group (10 mg/kg), RF-EMR + high dose of orientin group (20 mg/kg) and high dose of orientin group (20 mg/kg). All the groups except deionised water and high dose groups were exposed to 900 MHz radiofrequency-electromagnetic radiation for 28 consecutive days (1 h/day). Learning and memory was assessed via the step-down inhibitory avoidance task. Activities of lipid prexidation and antioxidant enzymes were measured using kits. Results: Radiofrequency electromagnetic radiation caused impairment in learning and memory and reduced activities of brain antioxidant enzymes, increased lipoperoxidation and corticosterone concentration as well as histopathological aberrations in the hippocampal tissues. Conversely, orientin alleviated learning and memory deficit, improved the activities of endogenous antioxidant enzymes and mitigated brain lipoperoxidation and neuronal degeneration in mice exposed to radiofrequency electromagnetic radiation. Conclusions: Orientin alleviates learning and memory impairment due to radiofrequency electromagnetic radiation in mice by improving antioxidant defence mechanism and may be considered as a promising therapeutic agent for improving the antioxidant system of people living in radiofrequency electromagnetic radiation-prone environment.
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Aim To study the influence of orientin on the cognitive function in APP/PS1 double transgenic mice and the probable mechanisms. Methods The mice (7 months of age) were randomly assigned into three groups ( n = 8 in each group) ; non-transgenic mice (NT) and APP/PS1 transgenic mice (Tg) were given saline, and APP/PS1 transgenic mice with orientin ( Tg + Ori) , and were injected intraperitoneally once per day for 30 days. Morris water maze test was carried out for the evaluation of spatial learning and memory. The measurement of A|3 was conducted by immunohistochemical staining. Autophagy related protein expressions of LC3-II , p62, Cathepsin D and Be-clin-1 were measured by Western blot. Results Compared with NT group, Tg mice showed deficit memory formation, enhanced A(3 deposition and protein levels of LC3-H , p62 and Cathepsin D; compared with Tg group, Tg + Ori mice showed improved learning and memory, reduced A(3 load, and decreased levels of LC3-FJ ,p62 and Cathepsin D. The expression of Bec-lin-1 showed no difference between groups. Conclusions Orientin could improve the cognitive function, reduce amyloid plaques and ameliorate hippocampus autophagic clearance of APP/PS1 transgenic mice.
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Objective To investigate the regulation effects of orientin on mitochondrial quality control in cardiomyocytes of rats with myocardial ischemia/reperfusion injury (MIRI). Methods A total of 75 Sprague-Dawley male rats were randomly divided into five groups: sham group, model group, and orientin preconditioning groups (1.0, 2.0, and 4.0 mg/kg). Except for sham group, all rats were exposed to modeling of MIRI with ligation of left anterior descending coronary artery (LADCA) for 30 min, and then releasing for 120 min. Mitochondrial membrane potential (MMP) and opening of mitochondrial permeability transition pore (mPTP) were detected by Microplate Reader; Activity of mitochondrial respiratory chain complexes I-IV was detected by Spectrophotometer; Cardiomyocyte apoptosis was detected by TUNEL method; Apoptosis and mitophagy related proteins were detected by Western Blot and the interaction between Parkin and p62 on mitochondria were detected by Co-IP. Results Orientin significantly restored MMP, mPTP opening threshold and activity of mitochondrial respiratory chain complexes I-IV, and reduced cardiomyocyte apoptosis. Additionally, orientin can dramatically inhibit the protein expression level apoptosis and mitophagy related proteins and weaken the interaction between Parkin and p62. Conclusion Orientin has a significant protective effect on cardiomyocyte mitochondria in MIRI rats. Its underlying mechanism is related to inhibition of hyperactivity of mitophagy in cardiomyocytes via Parkin-dependent and Parkin-independent signaling pathways.
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Objective: To determine the contents of five flavonoids in Spirodelae Herba from different growing areas, and evaluate its anti-oxidant activity. Methods: Contents of orientin, vitexin, cynaroside, apigenin-7-glucoside, and luteolin in 17 batches of Spirodelae Herba from eight origins were determined by HPLC with acetonitrile and 0.1% formic acid as mobile phase, and DPPH method was used to compare their anti-oxidant activities, together with cluster analysis and principal component analysis, the quality of Spirodelae Herba was evaluated. Results: The contents of five flavonoids in Spirodelae Herba from different growing areas exist certain differences, which can be divided into three different categories by cluster analysis and principal component analysis. The contents of orientin, vitexin, and cynaroside were significantly higher in samples from Shaanxi and Shanxi provinces than those from other origins, as well as their oxidation clearance rates were relatively high. The correlation analysis showed there was a significant positive correlation between the rate of oxidative elimination and flavonoid content. Conclusion: Spirodelae Herba from Shaanxi and Shanxi provinces has higher contents of flavonoids with strong antioxidant capacity. The analytical method is stable and reliable, which can provide the reference for enhancing the quality control of Spirodelae Herba.
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Aim To investigate the effect of orientin on proliferation and differentiation of 3T3-L1 pre-adipocytes and on insulin resistance(IR) in 3T3-L1 adipocytes and the possible mechanisms.Methods MTT assay and oil red O staining were applied to investigate the proliferation and the differentiation of 3T3-L1 pre-adipocytes, respectively.The intracellular triglyceride(TG) contents were detected by enzymatic analysis.IR model was induced with dexamethasone.A fluorescent glucose analogue, 2-NBDG, was used to measure the rate of glucose uptake.Western blot was used to detect the protein level of GLUT4 and phosphorylation of AMPK and ACC.The GLUT4 translocation was measured by fluorescent-immunohistochemistry.Results Orientin decreased the formation of lipid droplets and intracellular TG contents(P<0.01) in a concentration-dependent manner(P<0.05), but it had no obvious effects on the cell vitality.Under the IR state, orientin significantly increased 3T3-L1 adipocytes glucose uptake(P<0.05).Meanwhile, orientin up-regulated the protein expression of p-AMPK, p-ACC, and enhanced GLUT4 translocation and its expression.Conclusion Orientin can effectively inhibit the differentiation of 3T3-L1 pre-adipocytes and increase insulin sensitivity due to the activation of AMPK/GLUT4 signal pathway.
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Aim To investigate the role of autophagy in the cardioprotection by orientin and the relative molec-ular mechanisms in cell apoptosis and autophay. Meth-ods The isolated ischemia reperfusion ( I/R ) heart model was built firstly. The Experiments were divided into seven groups:control group, I/R group, different concentrations of orientin-treated group ( 1. 0 mg · kg-1 ,2. 0 mg· kg-1 ,4. 0 mg · kg-1 ) , autophagy in-hibitor group and resveratrol group. Hemodynamic in-dex were recorded by PowerLab, the activities of myo-cardial enzymes were detected through Biochemical an-alyzer, the ultrastructure changes and autophagosomes in myocardial cells were detected by electron microsco-py, the apoptosis was detected by TUNEL, and LC3 and Beclin1 protein levels of left ventricle were meas-ured by Western blot. Results Orientin at middle and high concentrations(2 and 4 mg·kg-1 ) induced auto-phagy shown by increased the number of autophago-somes, LC3-Ⅱ/LC3-Ⅰ ratio and Beclin 1 expression after ischemia-reperfusion. The induction of autophagy by orientin was correlated with enhanced cardiac func-tion and decreased apoptosis. But wortmannin, a kind of autophagy inhibitor, significantly attenuated the ori-entin-induced autophagy and increased apoptosis. Con-clusion The cardioprotection of orientin against myo-cardial ischemia reperfusion injury may be mediated through the inhibition of apoptosis and induction of au-tophagy.
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Objective: To study the pharmacokinetics (PK) and tissue distribution of orientin in rabbits. Methods: The high-performance liquid chromatography method was developed and validated for the determination of orientin in rabbit plasma and tissues. The PK characteristics of orientin in rabbits were compared by three kinds of administration, iv, im, and ip injection, using 3P97 pharmacokinetic software to examine the PK parameters. Results: The elimination half-life (t1/2β) of three kinds of administration was higher than the distribution half-life (t1/2α). The PK of orientin eliminating in rabbit fitted the two compartment model. Orientin was mainly eliminated in rabbit. The distribution of orientin in tissue after iv injection was investigated, and the order of orientin AUC in each group was kidney > liver > lung > spleen > heart > brain. Conclusion: Orientin is distributed extensively in tissues such as kidney, liver, and lung. It is difficult for orientin to cross the blood brain barrier. © 2013 Tianjin Press of Chinese Herbal Medicines.
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Objective: To establish a method for preparation of orientin and isoorientin from bamboo leaf flavones (BLF) using preparative high-performance liquid chromatography (PHPLC). Methods: After BLF was purified by AB-8 resin-based column chromatography, PHPLC was employed to isolate orientin and isoorientin, using a mobile phase consisting of methyl alcohol-water with 0.3% acetic acid (32:68). The flow rate was 5 mL/min while the column temperature was maintained at 20℃. The injection volume was 400 μL and the concentration of sample was 15.3 mg/mL. Signal was monitored at 330 nm with the diode array detector. Results: HPLC analysis showed that the purities of isoorientin and orientin were >99% and >93%, respectively, and the recovery rates were (92.7±2.92)% and (85.1±3.45)%, respectively. The confirmation of chemical structures was performed by 1H-NMR and 13C-NMR. Conclusion: The method is simple, with low toxicity but high recovery rate, and can be applied to the separation and preparation of isoorientin and orientin.
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Objective: To study the chemical compounds from the leaves of Cajanus cajan. Methods: Normal phase silica gel, medium-pressure ODS, MCI, and Sephadex LH-20 gel column chromatographies were used to isolate and purify the constituents. And their structures were identified by their physical properties, nuclear magnetic resonance spectrum, mass spectrometry, etc. Results: Twenty-three compounds were separated from the alcohol extract in the leaves of C. cajan. They were cajanine (1), longistyline A (2), longistyline C (3), cajanolactone A (4), pinostrobin (5), orientin (6), isovitexin (7), vitexin (8), cajanol (9), cajanin (10), prunetin (11), pratensein (12), (2R, 3R)-2, 3-dihydro-5-7, 4'-dimethoxyflavone (13), ethyl 10', 16'-dihydroxy hexadecanoate (14), vanillic acid (15), ethyl heptadecanoate (16), 2-O-quebrachitol (17), 2, 3, 4-trihydroxy-isovaleric acid (18), stigmasterol (19), betulinic acid (20), heptadecanoic acid (21), β-sitosterol (22), and β-daucosterol (23). Conclusion: Compounds 10-19 are obtained from this plant for the first time.
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Objective: To explore the extraction process of Trollii Flos Oral Liquid (TFOL). Methods: HPLC was used to determine orientin and vitexin in liquid extract of TFOL, using mass concentration of both as index. The orthogonal design L18(37) and reverse continuous cycle cryogenic extraction were used to extract Trollii Flos, the optimum extraction was determined, compared with the current standard of TFOL extraction process. Results: The optimum extraction conditions of reverse continuous cycle cryogenic extraction were as follows: extraction temperature 90°C, extraction time 2 h, powder type 10 mesh, added of 20 times water. The linear ranges of orientin and vitexin were 18.46-295.37 (r = 0.9999) and 5.84-93.44 μg/mL (r = 0.9999), the average recoveries of orientin and vitexin were 96.07%, RSD = 0.74% (n = 6), and 97.73%, RSD = 1.42% (n = 6), respectively. Conclusion: The technology of reverse continuous cycle cryogenic extraction is stable and feasible, and the extraction rate is high; The measurement method is accurate, sensitive, simple, and stability, and has good reproducibility. The extraction and determination methods could be used as production process and quality control reference of TFOL.
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OBJECTIVE@#To evaluate in vitro antioxidant and apoptotic activities of Cyperus rotundus (C. rotundus).@*METHODS@#The phytochemical study and the antioxidant activities of both methanol and aqueous extracts from C. rotundus aerial part were determined. In addition, these extracts were also investigated for their cytotoxic and apoptotic activities. The major compound of the methanol extract was isolated. Both methanol and aqueous extracts (300, 150, and 50 μg/mL) were evaluated for their antioxidant activity by the xanthine/xanthine oxidase assay system. However, 16, 8, and 4 mg/mL of each extract were tested to investigate their OH. formation scavenging potential. Aqueous extract (800, 400, and 200 μg/mL) and methanol extract (350, 175, and 88 μg/mL) were tested against lipid peroxidation, induced by 75 μM H2O2. The cytotoxicity (by MTT assay) and cell DNA fragmentation of both extracts were evaluated towards K562 and L1210 cell lines. The major compound was obtained from the butanol fraction of methanol extract and its structure was determined by RMN spectroscopic analysis.@*RESULTS@#The methanol and aqueous extracts showed respectively, 88% and 19% inhibition of xanthine oxidase activity. Yet, the same extracts inhibited lipid peroxidation by 61.5% and 42.0%, respectively. Both extracts inhibited OH. formation by 27.1% and 25.3%, respectively. Only methanol extract induced DNA degradation. Orientin was determined as the major compound isolated from the butanol fraction of methanol extract.@*CONCLUSIONS@#It appears that C. rotundus extracts exhibit a potential use as a natural antioxidant and an apoptosis inducer.
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Humans , Antioxidants , Chemistry , Metabolism , Pharmacology , Apoptosis , Cell Proliferation , Cyperus , Chemistry , Flavonoids , Glucosides , K562 Cells , Lipid Peroxidation , Plant Extracts , Chemistry , Pharmacology , Polyphenols , Chemistry , Pharmacology , Xanthine Oxidase , MetabolismABSTRACT
OBJECTIVE: To develop a method for the determination of orientin, vitexin, hyperin, neomangiferin, mangiferin, timosaponin BII and timosaponin AIII in Jinlianqingre granules(traditional Chinese medicines) by HPLC-MS. METHODS: The samples were separated on a Diamonsil C18 column (4.6 mm × 150 mm, 5 μm) by gradient elution using acetonitrile and 0.1% aqueous formic acid as the mobile phase at a flow rate of 800 μX · min-1. The column temperature was 30°C. Multiple-reaction monitoring (MRM) scanning was employed for quantification with switching electrospray ion source polarity between positive and negative modes in a single run. RESULTS: Complete separation was obtained within 15 min for the seven compounds. The regression equations showed linear relationships between the peak area and content of each compound (r > 0.9992). The average recoveries of the compounds listed above were 99.34%, 99.28%, 101.6%, 101.3%, 100.5%, 101.9% and 98.86% (n = 3), and the RSDs were 1.12%, 1.36%, 1.57%, 1.74%, 1.52%, 1.43% and 0.92%, respectively. CONCLUSION: The method is simple, accurate and highly reproducible, and can be used for the determination of seven compounds in Jinlianqingre granules.
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Objective To develop a HPLC method for quantitative determination of orientin and vitexin in Jinlianhua Pills and to provide valuable data for quality control of Jinlianhua Pills.Methods The separation was performed on a Shimadzu VP-ODS column (250 x 4.6mm,5μm)at 35℃ using a mobile phase of 0.5％ acetic acid solution-acetonitrile(88:12).The detection wavelength was 340nm.Results The calibration curve of orientin showed a good linearly within test ranges of 25.0 ～ 75.0μg/ml.The average recovery was 99.9％ with RSD 1.2％.The calibration curve of vitexin showed a good linearly within test ranges of 8.0 ～ 24.0μg/ml.The average recovery was 100.5％ with RSD 1.3％.Conclusion The developed method is simple,accurate and with good repeatability.It is suitable for quality evaluation of Jinlianhua Pills.
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AIM@#To provide a comprehensive procedure to evaluate the contribution of the floral parts to the yield of the major components from the flowers of Trollius chinensis, to underlay the selective breeding, cultivation, development, and utilization of the flowers.@*METHODS@#Five floral parts from eleven batches of the flowers of T. chinensis were examined by HPLC analysis for the content of orientin and vitexin, and by gravimetric analysis for their respective mass fraction. The contribution of each floral part was calculated using mathematical methods based on the results of the content and mass fraction. Variance analysis was carried out by Kruskal-Wallis H test and PCA method.@*RESULTS@#The calculated mean contributions of calyx, corolla, stamens and pistils, stalk, and ovary to the yield of both orientin and vitexin were 76.99% and 71.93%, 9.60% and 8.33%, 9.21% and 8.10%, 2.17% and 6.62%, and 2.03% and 5.02%, respectively.@*CONCLUSION@#The floral parts contribute unequally to the yield of orientin and vitexin, and the calyx contributes the highest and makes a significant difference compared with any other part.
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Apigenin , Drugs, Chinese Herbal , Flavonoids , Flowers , Chemistry , Glucosides , Ranunculaceae , ChemistryABSTRACT
AIM: To determine the contents of orientin,isoorientin,isovitexin in Bamboo Leaf from different sources. METHODS : Orientin,isoorientin,isovitexin were determined by HPLC. The mobile phase was MeOH-H_2O-HAc (35∶65∶1). The detective wavelength was at 340nm. RESULTS : The highest content of three favonoids was 0.89% and the lowest content was 0.07% from different sources. CONCLUSION : This method is simple,quick and accurate for determination of the three favonoids in Bamboo Leaf from different sources.