Rutin forestalls dysregulated cardiac bioenergetics in bisphenol A and dibutyl phthalate-exposed rats through PPAR? and AMPK modulation

Objectives: Proper cardiac function is greatly dependent on adequate supply and metabolism of energy substrates. Environmental pollutants exposure including plasticizers can trigger adverse cardiac metabolic events. This study was designed to investigate the ameliorative effect of rutin (Rt) on dysregulated cardiac energy metabolism in plasticizer-exposed rats. Materials and Methods: Forty-two rats were randomised into seven groups (n = 6): Control (0.1% dimethyl sulfoxide), bisphenol A (BPA, 25 mg/kg, p.o), dibutyl phthalate (DBP, 25 mg/kg, p.o), BPA + Rt 25 mg/kg, Rt 50 mg/kg, DBP + Rt (25 mg/kg, Rt 50 mg/kg), BPA + DBP and BPA + DBP + Rt, daily for 21 days. Results: BPA and DBP exposure increased plasma glucose, reduced insulin, and increased plasma and cardiac free fatty-acid. Cardiac glucose-6-phosphate level, hexokinase and pyruvate dehydrogenase activities increased in DBP while BPA reduced these variables. Cardiac glucose transporter-4 expression was reduced in BPA group, while cardiac peroxisome proliferator-activated receptor-alpha (PPAR?) and AMP-activated protein kinase (AMPK) expression increased in BPA and DBP-treated rats. However, Rt administration prevents impaired cardiac bioenergetics and glucometabolic regulation. Conclusion: Summarily, Rt improves BPA and DBP-impaired cardiac bioenergetics through PPAR? and AMPK modulation.

Rutin Promotes Browning of 3T3-L1 Preadipocytes and Its Mechanism / 中国实验方剂学杂志

ObjectiveTo investigate the effect of rutin on the browning of 3T3-L1 preadipocytes and the mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of different concentration of rutin （3.125， 6.25， 12.5， 25， 50， 100， 200 μmol·L-1） on 3T3-L1 cell activity， and Western blot to examine the effect of rutin （12.5， 25， 50 μmol·L-1） on the expression of thermogenesis-associated proteins uncoupling protein 1 （UCP1）， PR domain containing 16 （PRDM16） and peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α） in adipocytes. After the optimal concentration of rutin was determined， the effect of rutin on lipid droplet formation in adipocytes was observed based on oil red O staining， and the expression of nuclear respiratory factor 1 （NRF1）， nuclear respiratory factor 2 （NRF2） and mitochondrial transcription factor A （TFAM）， which were the landmark proteins of mitochondrial biosynthesis， was detected by Western blot. ResultCompared with the blank group， 200 μmol·L-1 rutin inhibited 3T3-L1 cell activity （P<0.01）. Compared with the blank group， at the concentration of 12.5， 25， 50 μmol·L-1 rutin significantly promoted the expression of thermogenesis-associated proteins （UCP1， PRDM16， and PGC-1α） （P<0.01）， which was determined as the optimal concentration. Compared with the blank group， 50 μmol·L-1 rutin significantly increased the immunofluorescence intensity of mitochondrial UCP1 protein in 3T3-L1 cells （P<0.01） and the expression of the markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM） （P<0.01）. In addition， 50 μmol·L-1 rutin significantly inhibited lipid droplet formation of 3T3-L1 adipocytes （P<0.01）. ConclusionRutin inhibited lipid droplet deposition in 3T3-L1 adipocytes and increased the expression of thermogenesis-related proteins （UCP1， PRDM16， and PGC-1α） and markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM）， thereby inducing the browning of 3T3-L1 adipocytes. This lays a basis for the development of drugs that safely regulate the browning of white cells.

Simultaneous determination of five components in Forsythia suspensa flower with UPLC-QAMS / 中国药房

OBJECTIVE To establish the methods for simultaneous determination of rutin， forsythiaside A， （+）-pinoresinol-4- O-β-D-glucopyranoside， forsythin and forsythigenin in Forsythia suspensa flower. METHODS UPLC method was adopted. The determination was performed on ACQUITY UPLC HSS T3 C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at the flow rate of 0.3 mL/min. The detection wavelengths were set at 275 nm （0-8 min），330 nm （8-10.5 min），275 nm （10.5-32 min）， respectively. The column temperature was 25 ℃， and sample size was 1 μL. Taking rutin as reference， the content of each component was determined by quantitative analysis of multi-components by single-marker （QAMS） method， and then compared with external standard method. RESULTS The contents of forsythiaside A， （+）-pinoresinol-4-O-β-D- glucopyranoside， forsythin and forsythigenin by QAMS were 7.472-7.671， 2.919-2.986， 1.439-1.486， 1.523-1.566 mg/g； the results obtained by the external standard method were 7.454-7.664， 2.913-2.996， 1.444-1.484， 1.519-1.562 mg/g， respectively. There was no significant difference in the measurement results between the two methods， with a relative deviation less than 1.0%. CONCLUSIONS This study successfully establishes the UPLC-QAMS method for simultaneous determination of five components in F. suspensa flower， and the results obtained by this method are not significantly different from those obtained by the external standard method. It can be used for quality control of F. suspensa flower.

Therapeutic potential of rutin in male infertility: A mini review / 中西医结合学报

Male infertility has become a problem worldwide, and recent research has emphasized the development of more effective therapy options. Among natural compounds, rutin has been widely studied for its potential to treat dysfunction related to male infertility, including a reduction in sperm quality, spermatogenesis disruption and structural disruption in the testis. A thorough review of scientific literature published in several databases, including Google Scholar, PubMed/MEDLINE and Scopus, was used to synthesize the present state of research on the role of rutin in male reproductive health. Rutin has been shown to possess antiapoptotic, antioxidant and anti-inflammatory activities, among others, which are crucial in the management of male infertility. Numerous investigations have shown that rutin protects against male infertility and have explored the underlying mechanisms involved. The present review, therefore, assesses the therapeutic mechanisms involved in male infertility treatment using rutin. Rutin was able to mitigate the induced oxidative stress, apoptosis, inflammation, and related physiological processes that can cause testicular dysfunction. Please cite this article as: Rotimi DE, Elebiyo TC, Ojo OA. Therapeutic potential of rutin in male infertility: A mini review. J Integr Med. 2023; 21(2): 130-135.

La mutación N15Y en la proteína E del coronavirus 2 del síndrome respiratorio agudo severo afecta la unión de doxiciclina y rutina / The N15Y mutation in the E protein of severe acute respiratory syndrome coronavirus 2 affects the binding of doxycycline and rutin

The severe acute respiratory syndrome coronavirus 2 genetic variation must be closely monitored. Viral transmission can inevitably result from mutations in the viral genome and functional proteins that aid in the virus's adaptation to the host. This study aimed to look for mutations in the E protein and see how they affected the ligands' molecular docking. Mathematical saturation mutagenesis and other informatics techniques were used. Fourteen severe acute respiratory syndrome coronavirus 2 isolates from Iraq were selected. Doxycycline and rutin were chosen as ligands. In four strains of severe acute respiratory syndrome coronavirus 2, the N15Y mutation was detected in the envelope protein. Depending on the calculation of the amount of energy of the atoms, this mutation is critical in modifying the shape of the protein as well as increasing protein stability. In the single-chain mutation, one pocket was determined, while all pentamer chains had two pockets. The N15Y mutation altered the degree of doxycycline binding by affecting the residue of attachment of the ligands. It also altered the position of the rutin's attachment to the E protein, which has a clear impact on the virion particle(AU)

La variación genética del coronavirus 2 del síndrome respiratorio agudo severo debe ser monitoreada de cerca. La transmisión viral puede resultar inevitablemente de mutaciones en el genoma viral y proteínas funcionales que ayudan en la adaptación del virus al hospedero. Este estudio tuvo como objetivo buscar mutaciones en la proteína E y ver cómo afectaban el acoplamiento molecular de los ligandos. Se utilizó la mutagénesis de saturación matemática y otras técnicas informáticas. Se seleccionaron 14 aislamientos del coronavirus 2 del síndrome respiratorio agudo severo en Irak. Se eligieron doxiciclina y rutina como ligandos. En cuatro cepas del coronavirus 2 del síndrome respiratorio agudo severo, se detectó la mutación N15Y en la proteína de la envoltura. Dependiendo del cálculo de la cantidad de energía de los átomos, esta mutación es fundamental para modificar la forma de la proteína y aumentar la estabilidad de la proteína. En la mutación de cadena única, se determinó un bolsillo, mientras que todas las cadenas de pentámero tenían dos bolsillos. La mutación N15Y alteró el grado de unión de doxiciclina al afectar el residuo de unión de los ligandos. También alteró la posición de unión de la rutina a la proteína E, lo que tiene un claro impacto en la partícula del virión(AU)

Systemic therapy with bromelain-trypsin-rutoside combination in inflammation: A narrative review of the pharmacodynamics

Inflammation and wound healing are attended by interactions of numerous cell types, mediators, cytokines, and the vascular system. Any imbalance in these interactions can set off a cascade of processes, often working in different directions, and which continue till homeostasis is restored. Systemic enzyme therapy with trypsin-bromelain-rutoside combination has long been used as dietary supplements or pharmaceuticals commercially to counter various inflammatory conditions. The clinical benefits are believed to be due to the multipronged action of the combination, which include anti-kinin, antioxidant, antiplatelet, anti-chemokine, pro-fibrinolytic, and pro-fibrotic activities; with more recent evidence indicating inhibition of pro-inflammatory signal transduction. The primary modes of action of the individual agents on different components and processes involved in inflammation have been evaluated and elucidated, using various in vitro, animal model, and clinical studies. A narrative review of the evidence collected over the years has been provided.

In vitro evaluation of free flavonoids in Cassia fistula Linn. calli of different ages under varying concentrations of plant growth regulators (PGRs)

Cassia fistula Linn, a paramount medicinal plant consumed by huge population all over the world. Its bioactive metabolites include flavonoids, phenolics, and polyphenols. Major objective of current study is to find right media combination and age of callus for maximum accumulation of free flavonoids in Cassia fistula cultures. In this study, the calli were induced in MS + 1.0mg/L 2,4-D + 0.05 mg/L BAP with 50 mg/L ascorbic acid. These calli were then transferred to basic MS medium with 1.0 mg/L2,4,5-T and varied BAP concentrations. Four and 8 weeks old calli were used to find out the right concentration of BAP and age for upscaling of free flavonoids. The samples were extracted and HPTLC analyzed for free flavonoids at 260 nm along with the leaf explants. The study revealed that the amount and number of free flavonoids in 4 weeks old calli increased as the amount of cytokinin (0.5 to 1.0 mg/L BAP) increased but decline with further addition of BAP (2.0 mg/L BAP). In aged calli number and amount of free flavonoids exhibit a tremendous increase when compared to the calli in the same medium at 4 weeks. This is inferred that accruing age of callus, reduced quantities of plantgrowth regulators (optimized for callus growth) accumulate free flavonoids. The best medium for free flavonoid production is MS+ 1.0 mg/L 2,4,5-T with 1.0 mg/L BAP may be recommended for producing a high amount of free flavonoids in C. fistula in vitro.The results were justified with the results of quantification.

Antiproliferative and anticancer activities of Neurada procumbens L. against epithelial carcinoma and breast cancer cell lines

Neurada procumbens L. commonly called as sand button, is used as a medicinal herb by Bedouin in the Arabia Peninsula for heart and respiratory functions. This little known plant has not been investigated in detail for its biological activities. Hence, we have made an attempt to investigate the plant for its anticancer activities. Methanol and chloroform extracts of N.procumbens were evaluated for antiproliferative and anticancer activities in HEp-2 and MCF-7 cancer cell lines. Humanforeskin epithelial cells showed an IC50 value of 34 ?g and 149 ?g for chloroform and methanol extracts, respectively. Thechloroform extract of N. procumbens @3 ?g showed 50% reduction of HEp-2 cell population in 24 h, while 5.5 ?g showedsimilar effects in MCF-7 cells. The results of the methanol extracts were little varied, where 49 ?g showed slow inhibitionof HEp-2 cells over a period of 72 h. In another similar observation, 76 ?g of methanol extract inhibited 50% MCF-7 cellproliferation. The concentrations which could inhibit complete turnover of the experimental cancer cell lines at 24 h were>5 ?g and 6 ?g of chloroform extract and > 60 ?g and 80 ?g of methanol extract for HEp-2 and MCF-7, respectively. Themicro therapeutic index of methanol extract in HEp-2 and MCF-7 were 3 and 1.9, while chloroform extract was 11.3 and6.1. This is an initial observation of N. procumbens that shows antiproliferative activities against cancer cell lines.

Highly sensitive electrochemical determination of rutin based on the synergistic effect of 3D porous carbon and cobalt tungstate nanosheets / 药物分析学报

Rutin,a flavonoid found in fruits and vegetables,is a potential anticancer compound with strong anti-cancer activity.Therefore,electrochemical sensor was developed for the detection of rutin.In this study,CoWO4 nanosheets were synthesized via a hydrothermal method,and porous carbon(PC)was prepared via high-temperature pyrolysis.Successful preparation of the materials was confirmed,and character-ization was performed by transmission electron microscopy,scanning electron microscopy,and X-ray photoelectron spectroscopy.A mixture of PC and CoWO4 nanosheets was used as an electrode modifier to fabricate the electrochemical sensor for the electrochemical determination of rutin.The 3D CoWO4 nanosheets exhibited high electrocatalytic activity and good stability.PC has a high surface-to-volume ratio and superior conductivity.Moreover,the hydrophobicity of PC allows large amounts of rutin to be adsorbed,thereby increasing the concentration of rutin at the electrode surface.Owing to the syn-ergistic effect of the 3D CoWO4 nanosheets and PC,the developed electrochemical sensor was employed to quantitively determine rutin with high stability and sensitivity.The sensor showed a good linear range(5-5000 ng/mL)with a detection limit of O.45 ng/mL.The developed sensor was successfully applied to the determination of rutin in crushed tablets and human serum samples.

RP-HPLC simultaneous quantification of rutin, avobenzone, and octyl methoxycinnamate in the presence of hydroxypropyl β-cyclodextrin (HPβCD) and sulfobutyl ether β-cyclodextrin (SBEβCD)

Abstract Development and validation of a simple and fast method of high-performance liquid chromatography with diode array detection (HPLC-DAD) for the simultaneously analysis of rutin, avobenzone, and octyl p-methoxycinnamate is presented. These substances were separated using a Kromasil C18 (250×4.6 mm, 5 μm) column, methanol: water (88:12 v/v) as the mobile phase, and a flow rate of 0.8 mL min−1. The experiment was performed at room temperature and elution was under isocratic conditions. Quantification was performed by external calibration at the wavelength of 325 nm. The validated parameters included linearity, selectivity, precision (repeatability), intermediate precision, accuracy, limit of detection, limit of quantification and robustness. The results of validation were statistically treated using the Action Stat version 3.5.152.34. The selectivity was also evaluated in the presence of two cyclodextrins (2-hydroxypropyl-β-cyclodextrin and β-cyclodextrin sulfobutyl ether sodium). The absence of parallelism between the curves of octyl p-methoxycinnamate in the absence and presence of the β-cyclodextrin sulfobutyl ether sodium in the mobile phase revealed interference from this matrix, thereby indicating the necessity of validating the method in the presence of this, and other matrices. The proposed method was selective, linear, precise, accurate, and robust for the simultaneous determination of rutin, avobenzone, and octyl p-methoxycinnamate.

Antitumor Effect and Mechanism of Rutin： A Review / 中国实验方剂学杂志

The morbidity and mortality of cancer have been on the rise， making it atop the list of human health threats. It has been a conundrum of global magnitude. As the side effects of chemotherapeutics seriously affect the life quality of cancer patients， it is urgent to find effective anti-cancer drugs with small toxic and side effects. In recent years， the anti-cancer effects of traditional Chinese medicine have attracted the interest of scholars. Owing to the improvement of medical research， an increasing number of anti-cancer components with small toxic and side effects have been extracted from traditional Chinese medicine. Rutin， a unique flavonoid in Chinese medicinals and many plants， proves to inhibit the proliferation of breast cancer， colon cancer， lung cancer， and prostate cancer cells. In addition， rutin alone or in combination with other therapeutic drugs can regulate a variety of signaling pathways and signal mediators of inflammation， apoptosis， autophagy， and angiogenesis， thereby suppressing tumor progression. Moreover， it can also alleviate the drug resistance of tumors and the side effect of chemotherapeutics. Nevertheless， it is limited by the low bioavailability and low solubility， to which nano delivery system turns to be a solution. At the moment， the anti-cancer potential of rutin and the molecular targets of it against various cancers have not been summarized and comprehensively analyzed. Therefore， the authors retrieved articles on the anti-cancer effects of rutin in recent years， summed up the mechanisms and molecular targets， and discussed relevant drug delivery systems and the safety， aiming at laying a theoretical foundation for further development and application of the flavonoid.

Rutin inhibits the in vitro formation of advanced glycation products and protein oxidation more efficiently than quercetin

Introduction: The exacerbated generation of advanced glycation end products (AGEs) triggers the onset of diabetic complications associated with hyperglycemia. The search for natural bioactive compounds that can inhibit AGE formation has gained immense interest. Quercetin and its glycoside derivative, rutin, are powerful antioxidants. They have been studied due to their potential to mitigate the disturbances observed in diabetes; however, studies comparing their antiglycation effects are limited. The aim of the present study was to compare the in vitro antiglycation potentials of quercetin and rutin. Methods: The in vitro model system of protein glycation was applied using bovine serum albumin (10 mg/mL) incubated with glucose (0.5 M) in the absence or presence of aminoguanidine (1 mM, prototype anti-AGE agent), metformin (1 mM), quercetin (100, 50, or 12.5 µM), or rutin (100, 50, or 12.5 µM). Before initiating incubations (day 0) and after 10, 20, and 30 days, aliquots were assayed for fluorescent AGEs. Markers of amino acid oxidation (dityrosine, N'-formylkynurenine, kynurenine), protein carbonyl groups (PCO), and protein crosslink formation were assessed after 30 days. Results: Both quercetin and rutin inhibited the formation of AGEs and decreased the PCO levels in a concentration-dependent manner, and moreover, the effect of rutin was more prominent than that of quercetin. Quercetin and rutin also decreased the formation of amino acid oxidation products and protein crosslinks; the best effects were observed in incubations with rutin. Conclusion: Rutin exhibited the most potent antiglycation and antioxidant activities, which may be attributed to the minor occurrence of interactions between albumin and rutin, making rutinnoside more available to exert its effects.

The effect of mild hypothermia plus rutin on the treatment of spinal cord injury and inflammatory factors by repressing TGF-β/smad pathway

ABSTRACT Purpose To probe the mechanism of mild hypothermia combined with rutin in the treatment of spinal cord injury (SCI). Methods Thirty rats were randomized into the following groups: control, sham, model, mild hypothermia (MH), and mild hypothermia plus rutin (MH+Rutin). We used modified Allen's method to injure the spinal cord (T10) in rats, and then treated it with MH or/and rutin immediately. BBB scores were performed on all rats. We used HE staining for observing the injured spinal cord tissue; ELISA for assaying TNF-α, IL-1β, IL-8, Myeloperoxidase (MPO), and Malondialdehyde (MDA) contents; Dihydroethidium (DHE) for measuring the reactive oxygen species (ROS) content; flow cytometry for detecting apoptosis; and both RT-qPCR and Western blot for determining the expression levels of TGF-β/Smad pathway related proteins (TGF-β, Smad2, and Smad3). Results In comparison with model group, the BBB score of MH increased to a certain extent and MH+Rutin group increased more than MH group (p < 0.05). After treatment with MH and MH+Rutin, the inflammatory infiltration diminished. MH and MH+Rutin tellingly dwindled TNF-β, MDA and ROS contents (p < 0.01), and minified spinal cord cell apoptosis. MH and MH+Rutin could patently diminished TGF-β1, Smad2, and Smad3 expression (p < 0.01). Conclusions MH+Rutin can suppress the activation of TGF-β/Smad pathway, hence repressing the cellular inflammatory response after SCI.

Research progress on rutin derivatives / 中国中药杂志

Compounds derived from natural products present satisfactory efficacy in disease prevention and treatment. The use of chemical substances in plants to promote healthhas increasingly attracted people's attention. Rutin, a typical flavonoid, is mainly found in various vegetables, fruits and Chinese herbal medicines. As a natural antioxidant, it features many pharmacological activities, such as anti-inflammation, anti-virus, anti-tumor, and prevention and treatment of cardiovascular and cerebrovascular diseases. However, the low bioavailability and poor water solubility limit its clinical application. In view of this, its structure is optimized and modified to afford rutin derivatives with good solubility, high bioavailability, stable metabolism and small toxic side effects. So far, a large number of rutin ethers, esters, and complexes have been synthesized and undergone activity testing. This paper reviews the structural modification of rutin in recent years, and the obtained derivatives have excellent properties and significant biological activity.

Enzymatic synthesis of acylated quercetin 3-O-glycosides: a review / 生物工程学报

Quercetin 3-O-glycosides (Q3Gs) are important members of quercetin glycosides with excellent pharmacological activities such as anti-oxidation, anti-inflammation, anti-cancer and anti-virus. Two representatives of Q3Gs, rutin and troxerutin, have been developed into clinical drugs, demonstrating Q3Gs have become one of the important sources of innovative drugs. However, the applications of Q3Gs in food and pharmaceutical industries are hampered by its poor bioavailability. Of the known means, selective acylation modification of Q3Gs through enzymatic catalysis to obtain Q3G esters is one of the effective ways to improve its bioavailability. Herein, the enzyme-mediated acylation of Q3Gs were reviewed in details, focusing on the four tool enzymes (acyltransferases, lipases, proteases and esterases) and the whole-cell mediated biotransformation, as well as the effect of acylations on the biological activities of Q3Gs. Furthermore, the highly efficient synthesis and diversification of acylated site for Q3G esters were also discussed. Taken together, this review provides a new perspective for further structural modifications of Q3Gs towards drug development.

Effect of Tartary Buckwheat Boiled Noodles on Postprandial Blood Glucose Level and Its Active Components / 日本補完代替医療学会誌

Dry solid matter (rutin content: 51.6 mg/g; quercetin content: 72.2 mg/g) extracted from Tartary buckwheat boiled noodles using 70% methanol as the solvent was found to have α-glucosidase inhibitory activity. As for fractions fractionated by silica gel column chromatography, the fractions rich in quercetin and rutin showed remarkable α-glucosidase inhibitory activity. Tartary buckwheat boiled noodles used as samples in this study contained quercetin produced from rutin by the action of rutinase, suggesting that both rutin and quercetin contained were involved in the α-glucosidase inhibitory activity of the dry solid extract. Changes in postprandial blood glucose levels were compared for boiled noodles made from two types of buckwheat (i.e., Tartary buckwheat and common buckwheat), revealing that blood glucose elevation after eating Tartary buckwheat boiled noodles was suppressed. The blood glucose level 40 minutes after eating Tartary buckwheat boiled noodles was significantly low (p<0.05). It can be concluded that this might be caused by the α-glucosidase inhibitory activity of rutin (270.0 mg) and quercetin (330.5 mg), which correspond to a total amount of 935 mg of rutin equivalents, in the gastrointestinal tract. As a result, the digestion of carbohydrates contained in the samples consumed and their absorption by the intestine might be inhibited, resulting in the suppression of increases in blood glucose levels. The presence of a certain amount of quercetin was considered to be key to the suppression of blood glucose elevation. It is important to control rapid postprandial blood glucose increases to prevent diabetes from developing or becoming serious. This study suggests the potential for Tartary buckwheat boiled noodles to contribute to diabetes prevention.

Simultaneous Determination of 5 Components in the Classical Formula Huaihua San by HPLC Wavelength Switching Method / 中国药房

OBJECTIVE：To develop a method for simultaneous determination of 5 components in classical formula Huaihua san，including rutin ，naringin，neohesperidin，quercetin and pulegone. METHODS ：HPLC wavelength switching method was adopted. The determination was performed on Cosmosil C 18 column with mobile phase consisted of acetonitrile- 0.05％ phosphoric acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 257 nm for rutin ，283 nm for naringin and neohesperidin ，254 nm for quercetin ，252 nm for pulegone ，respectively. The column temperature was set at 30 ℃， and sample size was 10 μL. RESULTS：The linear range was 21.7-2 170 μg/mL for rutin，46-4 600 μg/mL for naringin，22.3- 2 230 μg/mL for neohesperidin，0.96-96 μg/mL for quercetin，2.7-270 μg/mL for pulegone（all r＞0.999），respectively. RSDs of precision，stability（24 h）and reproducibility tests were all lower than 2％（n＝6）. Average recoveries were 100.70％，99.31％， 101.10％，100.03％ and 99.63％（all RSD ＜2％，n＝9）. Among 3 batches of Huaihua san samples ，the contents of above 5 components were 20.055-22.615，25.557-27.806，11.428-13.250，0.350-0.478，2.372-4.011 mg/g，respectively. CONCLUSIONS ： Established method is simple ，accurate and reproducible ，and could be used for the simultaneous determination of 5 components in Huaihua san.

One step assay of the four plant ingredients in Yangxue Anshen syrup by HPLC method / 药学实践杂志

Objective To establish a HPLC method for simultaneous determination of quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in Yangxue Anshen syrup. Methods Waters symmetry C18 column (250 mm×4.6 mm, 5 μm) was used with 0.1% acetic acid (A) and methanol (B) as the mobile phase. Gradient elution was performed at a flow rate of 1.0 ml/min, 0-15 min, 95%-90%A; 15-35 min, 90%-70%A; 35-55 min, 70%-60%A; 55-85 min, 60%-50%A; 85-95 min, 10%A. The detection wavelengths were 256 nm and 320 nm. Column temperature was 30 ℃ and the injection volume was 10 μl. Results Quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside showed good linear relationship within the range of 10-300, 5.0-150.0, 5.0-150.0, 20.0-600.0 µg/ml(r≥0.9989), respectively. The average recovery was (96.75±1.41)%, (99.61±1.01)%, (97.18±1.96)% and(99.12±0.97)% (n=6), respectively. Conclusion The established method is simple, accurate and stable, which can be used for the simultaneous determination of 4 components in Yangxue Anshen syrup.

Content determination of five flavonoids in Tibetan medicine Rhododendron anthopogonoides by quantitative analysis of multi-components by single marker (QAMS) / 中国中药杂志

To establish a quantitative analysis of multi-components by single marker(QAMS) method for five flavonoids in Rhododendron anthopogonoides and verify its feasibility and applicability in the medicinal materials of R. anthopogonoides. With hyperoside as the internal reference, relative correction factors(RCF) of rutin, quercetin, quercitrin and kaempferol were established by high-performance liquid chromatography(HPLC) analysis. RCFs were used to calculate the content of each component, system durability and relative retention time. Simultaneously, QAMS and external standard method(ESM) were used to determine the content of five flavonoids in 12 batches of R. anthopogonoides from different origins. The results were statistically analyzed to verify the accuracy and feasibility. The fingerprints and cluster analysis data of R. anthopogonoides analyzed and discussed differences among the batches. According to the results, the RCFs of rutin, quercetin, quercetin and kaempferol in R. anthopogonoides were 1.242 6, 0.990 5, 0.535 0, and 0.781 3, respectively. The RCFs represented a good reproducibility under different experimental conditions. Besides, there was no significant difference between QAMS and ESM. Besides, the fingerprint and cluster analysis data showed the consistency between the classification and with the origin distribution of the herbs. In conclusion, the QAMS method shows a good stability and accuracy in the quality control of R. anthopogonoides.

Rutin attenuates early brain injury after subarachnoid hemorrhage in rats by protecting blood-brain barrier / 国际脑血管病杂志

Objective:To investigate the effect of rutin on blood-brain barrier in early brain injury after subarachnoid hemorrhage (SAH) in rats.Methods:A rat model of SAH was induced by puncturing the internal carotid artery. The rats were divided into a sham operation group, a model group and a rutin (50 mg/kg) group. Twenty-four hours after modeling, SAH score and neurological deficit score were conducted, and brain water content and Evans blue extravasation rate were detected in each group. Western blot analysis was used to detect the expression of claudin-5, occludin and zonula occluden (ZO)-1. TUNEL staining was used to detect neuronal apoptosis.Results:Compared with the sham operation group, the SAH score increased, the neurological deficit score decreased, and the brain water content and Evans blue extravasation increased in the model group. Rutin could significantly reduce the SAH score, increase neurological deficit score, and reduce brain water content and Evans blue exudation (all P<0.01). Western blot analysis showed that the expression of claudin-5, occludin and ZO-1 protein decreased in the model group, and the expression of claudin-5, occludin and ZO-1 protein increased significantly in the rutin group ( P<0.01). In addition, the number of TUNEL positive cells induced by SAH in the rutin group decreased. Conclusion:Rutin can play a protective role in early brain injury after SAH, and its mechanism may be associated with protecting the integrity of blood-brain barrier.