Determination of Mangiferin and Homomangiferin in the Leaves of 93 Kinds of Mango

[Objectives]To determine the content of mangiferin and homomangiferin in mango leaves by HPLC.[Methods]The mangiferin and homomangiferin were separated and determined by Elite Hypersil C18(5μm,4.6 mm ID×250 mm)chromatographic column.Acetonitrile-0.1%(V/V)phosphoric acid solution was used as the mobile phase for gradient elution,the flow rate was 1.0 mL/min,the detection wavelength was 258 nm,the column temperature was 30℃,and the injection volume was 5μL.[Results]There was a good linear relationship between mangiferin and homomangiferin in the range of 0.0254-0.5080μg/μL(r=0.9999)and 0.000960-0.019200μg/μL(r=0.9999),respectively.The average recovery rate(n=6)of mangiferin and homomangiferin in mango leaves was 101.7%(RSD=2.0%)and 101.0%(RSD=1.7%),respectively.[Conclusions]There were great differences in the content of mangiferin and homomangiferin in the leaves of different varieties of mango.The experimental results could provide a scientific basis for further development and utilization of mango leaf resources.

Synthesis and evaluation of novel analogues of mangiferin as potent antipyretic

Objective:To screen different analogues of mangiferin pharmacologically for antipyretic activity. Methods:The naturally occurring xanthone glycoside mangiferin was isolated by column chromatography from the elhanolic extract of stem bark of Mangifera indica.Mangiferin was further converted to 5-(N-phenylamino methyleno) mangiferin.5-(N-p-chlorophenylamino methyleno) mangiferin,5-(N-2-methyl phenylamino methyleno) mangiferin.5-(N-p-methoxy phenylamino methyleno) mangiferin.5-(N,N-diphenylamino methyleno) mangiferin,5-(N-α-napthylamino methyleno) mangiferin and 5-(N-4-methyl phenylamino methyleno) mangiferin analogues.The synthesized compounds were further screened for antipyretic activity along with mangiferin at a dose level of 100 and 200 tng/kg.Mangiferin and its analogues were characterized by melting point and R_f value determination and through spectral technique like UV,IR,and NMR spectral analysis.Results:The antipyretic activity of mangiferin as well as all analogues was found to be more significant in at higher dose ie.200 mg/kg which was depicted thmugh a decrease in rectal temperature up to 3 h.Conclusions:The antipyretic activity of mangiferin and its analogues may be attributed to inhibition in synthesis of TNF-αand anti-oxidant activity associated with amelioration of inflammatory actions of cytokines.

Mangiferin抑制多发性骨髓瘤恶性生物学特性与发挥抗癌效应机制的实验研究

目的:探讨纯中药提取物Mangiferin对多发性骨髓瘤(MM)细胞恶性生物学行为的影响,分析Mangiferin抗骨髓瘤效应的分子机制,为MM替代治疗提供实验依据。方法:不同浓度Mangiferin干预人MM细胞株U266、RPMI8226细胞后,CCK-8法检测细胞增殖,Annexin V/PI双染流式细胞术检测细胞凋亡,Western blot检测凋亡及相关信号通路蛋白的表达,实时荧光定量聚合酶链式反应(qRT-PCR)检测基质金属蛋白酶(MMP)、CXC趋化因子受体(CXCR)家族的表达变化。结果:Mangiferin可抑制U266、RPMI8226细胞增殖活性,并诱导其凋亡。当Mangiferin干预U266和RPMI8226细胞48 h后,U266和RPMI8226细胞中Bcl-2家族促凋亡蛋白Bax表达上调,并下调survivin、Bcl-xL蛋白的表达同时水解活化caspase-3以促进细胞凋亡,且显著下调U266细胞中Bcl-2蛋白表达以诱导细胞凋亡(P<0.05)。在Mangiferin干预MM细胞后,其不仅可增加肿瘤抑制因子p53的表达水平,同时通过抑制抗凋亡分子的表达并下调AKT、NF-κB磷酸化水平继而诱发MM细胞程序性死亡。Mangiferin干预后可明显下调U266细胞CXCR4、MMP2及MMP9的表达(P<0.05),对CXCR2、CXCR7以及MMP13的表达基本无影响(P>0.05);而干预RPMI8226细胞后可下调CXCR4、MMP9、MMP13的表达(P<0.01),对MMP2表达影响微弱,对CXCR2、CXCR7的表达基本无影响(P>0.05)。结论:Mangiferin能抑制MM细胞增殖并诱导其凋亡,其机制可能与抑制NF-κB信号通路激活并通过影响Bcl-2家族蛋白表达以及使MMP、CXCR家族核心成员表达受抑等有关。

Mangiferin,a natural xanthone,accelerates gastrointestinal transit in mice involving cholinergic mechanism

AIM： To investigate the effects of mangiferin on gas- trointestinal transit （GIT） in normal and constipated mice, together with the possible mechanism.METHODS： Intragastrically-administered charcoal mealwas used to measure GIT in overnight starved Swiss mice. In the first experiments, mangiferin （3 mg/kg, 10 mg/kg, 30 mg/kg, and 100 mg/kg, po） or tegaserod （1 mg/kg, ip） were administered 30 min before the char- coal meal to study their effects on normal transit. In the second series, mangiferin （30 mg/kg） was tested on delayed GIT induced by several different pharma- cological agonists （morphine, clonidine, capsaicin） or antagonists （ondansetron, verapamil, and atropine） whereas in the third series, mangiferin （30 mg/kg, 100 mg/kg and 300 mg/kg） or tegaserod （1 mg/kg） were tested on 6 h fecal pellets outputted by freely fed mice. The ratio of wet to dry weight was calculated and used as a marker of fecal water content. RESULTS： Mangiferin administered orally significantly （P 〈 0.05） accelerated GIT at 30 mg/kg and 100 mg/kg （89% and 93%, respectively）, similarly to 5-hydroxytrypta- mine4 （5-H%） agonist tegaserod （81%） when compared to vehicle-treated control （63%）. Co-administered man- giferin （30 mg/kg） totally reversed the inhibitory effect of opioid agonist morphine, 5-HT3-receptor antagonist ondansetron and transient receptor potential vanilloid-1 receptor agonist capsaicin on GIT, but only to a partial extent with the GIT-delay induced by ~2-adrenoceptor agonist clonidine, and calcium antagonist verapamil. However, co-administered atropine completely blocked the stimulant effect of mangiferin on GIT, suggesting the involvement of muscarinic acetylcholine receptor activation. Although mangiferin significantly enhanced the 6 h fecal output at higher doses （245.5±10.43 mg vs 161.9±10.82 mg and 227.1±20.11 mg vs 161.9±10.82 mg of vehicle-treated control, at 30 and 100 mg/ kg, P 〈 0.05, respectively）, the effect of tegaserod was more potent （297.4±7.42 mg vs 161.9±10.82 mg of vehicle-treated control, P 〈 0.05）. Unlike tegaserod, which showed an enhanced water content in fecal pel- lets （59.20%±1.09% vs 51.44%±1.19% of control, P 〈 0.05）, mangiferin evidenced no such effect, indi-cating that it has only a motor and not a secretomotor effect. CONCLUSION： Our data indicate the prokinetic action of mangiferin. It can stimulate the normal GIT and also overcome the drug-induced transit delay, via a choliner- gic physiological mechanism.

Synthesis of mangiferin derivates and study their potent PTP1B inhibitory activity

Protein tyrosine phosphatase 1 B （PTP1 B） has received considerable attention from the drug industry as a potential treatment for diabetes mellitus. Mangiferin has been reported to possess significant antidiabetic activity. Based on the previous study, eight new mangiferin derivates were synthesized and evaluated for their PTP1B inhibitory activity. Some of them displayed good inhibitory activity on PTP1B.

Screening, Characterization and Evaluation of Mangiferin Polymorphs

Mangiferin is a compound with many pharmacological activities and exists in many natural products.Anhydrous and hydrate of mangiferin have been reported separately in two literatures,but the polymorphism of this compound has not been realized until this paper.In this study,polymorph screening of mangiferin has been carried out and five forms have been obtained including three new forms never reported.Several solid state characterization methods,such as powder X-ray diffraction,differential scanning calorimetry and thermogravimetry,are used to identify and characterize all of mangiferin forms.The comparison of the crystallographic data and hirshfeld surface analysis were first reported for mangiferin anhydrous and hydrate.Furthermore,the studies on stability,transformation and solubility have been undertaken,the results prompt that form V can be used as the dominant polymorph for the development of innovative pharmaceuticals.

Efficient Extraction and Isolation of Mangiferin from Mango Leaves by Ethyl Acetate Impurity Removal Method

[Objectives] To optimize the ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves,and provide raw materials and technical support for development and use of mangiferin related products. [Methods]Five steps( material crushing→ ethyl acetate impurity removing → concentrated extract washing → extracting with methanol → crystallization and precipitation) were used.The single factor experiment and L9( 34) orthogonal experiment was carried out to optimize the process parameters including extraction time,ultrasonic power,extraction times,and extraction temperature.[Results] The optimum process of ethyl acetate impurity removal method for extracting and isolating mangiferin from mango leaves was as follows: the mango leaves were crushed and sieved; 3 m L/g of ethyl acetate was added,sealed and soaked for 4 h,ultrasonically shaken for 20 min( 50℃,350 W),filtered at room temperature,filtered with 100 mesh sieve,and extracted three times; added 100% methanol to the residue at 3 m L/g,extract by ultrasonic vibration for 20 min( 350 W,55℃)for four times,filtered with 100 mesh sieve when it was still hot; mixed the extract of each time,condensed by vacuum decompression to get the extract; added 100% methanol at 4 m L/g,mixed and washed for 5 min at room temperature,placed for 10 min,filtered with 100 mesh sieve,washed 3 times repeatedly,and dried the filter residue at 60℃ to obtain the crude mangiferin; added 100% methanol at 4 m L/g,mixed and washed at 50℃ for 5 min,placed at 6℃ for 8 h,dried the filter residue at 60℃,and repeatedly crystallized two times. According to the above process,crude and pure mangiferin products could be obtained,the purity of mangiferin of the crude product was higher than 64. 00%,the total recovery rate was 83. 90%,and the purity of mangiferin of the pure product was higher than 98. 00%,and the total recovery rate was about 66. 40%. [Conclusions] The optimized ethyl acetate impurity removal method is easy in operation,low in cost,and high in efficiency for extracting and isolating mangiferin,and can be applied for actual production of mangiferin.

Mangiferin联合硼替佐米对Burkitt淋巴瘤恶性生物学行为的调控机制及对CXCR表达的影响

目的:分析芒果苷(mangiferin)联合硼替佐米对人Burkitt淋巴瘤Raji细胞增殖、侵袭、凋亡和自噬的作用及对CXC趋化因子受体家族(CXCR)表达的影响,并探究其间存在的分子机制,为Burkitt淋巴瘤基础研究与临床提供科学依据。方法:采用不同浓度Mangiferin、硼替佐米单药或联合干预Raji细胞,利用CCK-8法检测细胞增殖,Transwell小室法检测细胞侵袭能力,Annexin V/PI双染流式细胞术检测细胞凋亡,Western blot检测凋亡、自噬及Akt/m TOR通路蛋白表达情况,实时荧光定量PCR检测CXCR家族的表达变化。结果:不同浓度Mangiferin干预Raji细胞不同时间后,可抑制Raji细胞活力,且呈浓度及时间依赖性(r=-0.682,r=-0.836);与单药组相比,Mangiferin联合硼替佐米干预Raji细胞时,细胞增殖与侵袭能力显著下降、凋亡水平显著升高(P<0.01)。Mangiferin联合硼替佐米干预Raji细胞后,可上调促凋亡蛋白Bax表达并显著下调抗凋亡蛋白Bcl-2的表达,同时亦使Caspase-3水解活化,继而诱导Raji细胞发生凋亡。Mangiferin联合硼替佐米干预Raji细胞后可上调LC3Ⅱ蛋白的表达,且细胞中LC3Ⅱ/LC3Ⅰ比值较单药或对照组显著上调(P<0.01)。Mangiferin联合硼替佐米可显著抑制Akt和m TOR的磷酸化水平,通过抑制Akt/m TOR通路来使Raji细胞增殖及侵袭受抑,并诱导细胞发生自噬与凋亡。Mangiferin及硼替佐米单药干预Raji细胞后,可下调CXCR4、CXCR7 m RNA的表达,当两药联合时CXCR4、CXCR7 m RNA表达下调更为显著(P<0.01)。Mangiferin单药或联合硼替佐米干预Raji细胞后CXCR5 m RNA表达无显著变化(P>0.05),但两药联合时可使CXCR3表达下调(P<0.05)。结论:Mangiferin联合硼替佐米能协同抑制Raji细胞增殖、侵袭,并诱导其发生自噬与凋亡,机制可能与抑制Akt/m TOR信号通路并通过下调抗凋亡蛋白Bcl-2和上调促凋亡蛋白Bax以及使CXCR家族表达受抑等有关。

Pharmacokinetic Study and Tissue Distribution of Single Mangiferin After Intravenous Administration in Rats

A simple, sensitive ane selective high performance liquid chromatographic （HPLC） method with UV detection （320 nm） was developed and validated for determination of mangiferin in rat plasma and tissues. Mangiferin and internal standard （spinosin） were separated using mobile phase of acetonitrile-water （20：80, v/v） with 1% glacial acetic acid and 1% THF on a Phenomenex gemini C18 column. The flow rate was 0.7 mL/min. The calibration curves of mangiferin in plasma and tissues were linear over the investigated ranges. The intra- and inter-run preeisions for all samples were less than 13.8 %. The time-concentration curve of mangiferin after intravenous administration to rats corresponded to two-compartment model. The main pharmacokinetic parameters T0.5α, T0.5β, CL and AUC0-T were 15.87 min, 26.15 rain, 6.1 L/（min·kg） and 3.28 mg· min/mL, respectively. The highest and lowest levels of mangiferin occurred in spleen and brain, respectively. Mangiferin was not found in liver. After intravenous administration, the drug was distributed extensively and transferred quickly in rats in vivo.

Antiviral activity of mangiferin from the rhizome of Anemarrhena asphodeloides against herpes simplex virus type 1

Objective:To evaluate the antiviral activity of pure compounds against herpes simplex virus type 1(HSV-1)from the rhizome of Anemarrhena asphodeloides.Methods:Bioassay-guided isolation was conducted to separate the active compound and its chemical structure was elucidated by spectral analysis.In vitro antiviral efficacy of active compound was detected by Cell Counting Kit-8 assay,plaque reduction assay,and fluorescence observation.RT-PCR was used to determine the viral load and the cytokine-related gene expression after HSV-1 infection.In vivo study was also conducted to further determine antiviral efficacy of an active compound against HSV-1.Results:An active compound was isolated and elucidated as mangiferin.Mangiferin significantly inhibited the replication of HSV-1 in Vero cells with a half maximal inhibitory concentration(IC_(50))of 64.0 mg/L.Time-of-addition and time-of-removal assays demonstrated that mangiferin could effectively inhibit the replication of HSV-1 in the early stage(8 h).UL12,UL42,and UL54 gene expression levels of HSV-1 in the 64 mg/L mangiferin-treated group were markedly reduced as compared with the HSV-1 group(P<0.01).Fluorescence observation showed that mangiferin attenuated the mitochondrial damage maintainingΔΨm induced by HSV-1 in Vero cells.The expression of inflammatory factors TNF-α,IL-1β,and IL-6 was remarkably increased in the virus-infected group as compared with that in the normal group(P<0.05),the levels of these inflammatory factors dropped after treatment with mangiferin.Mangiferin significantly decreased the viral load and attenuated the HSV-1-induced up-regulation of TNF-α,IL-1β,and IL-6.The relative protection rate of HSV-1-infected mice could reach up to55.5%when the concentration of mangiferin was 4 g/kg.Conclusions:Mangiferin exhibits promising antiviral activity against HSV-1 in vitro and in vivo and could be a potential antiviral agent for HSV-1.

Mangiferin Promotes Bregs Level,Activates Nrf2 Antioxidant Signaling,and Inhibits Proinflammatory Cytokine Expression in Murine Splenic Mononuclear Cells In Vitro

Recent studies indicated that regulatory B cells(Bregs)and nuclear factor erythroid 2-related factor 2(Nrf2)antioxidant signaling pathway play important roles in the pathogenesis of chronic graft-versus-host disease(cGVHD).Mangiferin(MA),a polyphenol compound,has been reported to activate Nrf2/antioxidant-responsive element(ARE)signaling pathway.This study was aimed to investigate the effects of MA on Bregs and Nrf2 antioxidant signaling in murine splenic mononuclear cells(MNCs)in vitro.Our results revealed that MA could increase the Bregs level in murine splenic MNCs.Moreover,MA up-regulated the expression of Bregs-associated immunosuppressive factor interleukin-10(IL-10)by activating the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)and extracellular signal-regulated kinase(ERK)signaling in murine splenic MNCs.Meanwhile,MA inhibited the proinflammatory cytokines IL-2 and interferon-y(INF-y)at both mRNA and protein levels.MA also enhanced the transcription and protein expression of Nrf2 and NADPH quinine oxidoreductase 1(NQOl),whereas decreased that of Kelch-like ECH-associated protein 1(Keapl)in murine splenic MNCs.Moreover,MA promoted the proliferation and inhibited the apoptosis of murine splenic MNCs.These results suggested that MA exerts immunosuppressive effects by upregulating the Bregs level,activating the Nrf2 antioxidant pathway,and inhibiting the expression of pro-immunoinflammatory factors.MA,as a natural immunomodulatory and anti-inflammatory agent,may have a potential role in the prophylaxis and treatment of cGVHD.

Effects of Mangiferin on the Expression of TNF-α,iNos,ICAM-1 and Its mRNA in the Heart,Brain and Kidneys of SHR

[Objectives] To study the effects of Mangiferin( MGF) on TNF-α,iNOS,ICAM-1 and its mRNA expression in the heart,brain and kidneys of spontaneously hypertensive rats( SHR),and reveal the mechanism of its anti-inflammatory injury in hypertension target organs.[Methods]SHRs were randomly divided into 5 groups: the model group,the high-dose,medium-dose,low-dose MGF groups and the Benazepril group,with 8 rats in each group,WKY rats were used for the normal control group. Besides,on-invasive blood pressure( BP) instruments were used to measure systolic blood pressure in the rats' tail artery,western blot was used to analyze the expression of TNF-α,iNOS,ICAM-1 and reverse transcription-polymerase chain reaction( RT-PCR) was used to analyze the expression of TNF-α,iNOS,ICAM-1 mRNA.[Results]Compared with the normal control group,the model group's BP level was significantly increased( P <0. 01)),but the MGF had no significant lowering BP effect( P > 0. 05); compared with the normal control group,the expression of TNF-α,iNOS,ICAM-1 and its' mRNA in the model group was significantly increased( P < 0. 05 or P < 0. 01),and MGF could reduce the level of expression of these inflammatory cytokines( P < 0. 05 or P < 0. 01); between the Benazepril group and high-dose,medium-dose,low-dose MGF groups,most of the indicators had no significant difference( P > 0. 05). [Conclusions]MGF had no significant lowering BP effect,SHR showed inflammatory injury in the heart,brain and kidneys,MGF showed improvement on the inflammatory injury,and the anti-inflammation mechanism may be associated with lowering TNF-α,i NOS and ICAM-1 and its mRNA expression.

Adsorptive Stripping Voltammetric Determination of Mangiferin Using an Activated Chitosan Modified Carbon Paste Electrode

A medium molecular weight powdered chitosan modified carbon paste electrode was used to investigate the electrochemical behaviour by cyclic voltammetry of the pharmacologically-active ingredient mangiferin (MG). An irreversible system was observed, with a peak at ﹢0.55 V (vs Ag/AgCl). The peak current increases about fourfold, at the modified electrode in comparison with that recorded at the chitosan free carbon paste electrode. This allowed the use of adsorptive stripping voltammetry to develop a simple and sensitive electroanalytical method for the determination of MG. The influence of key parameters was investigated, including the electrolysis potential, the preconcentration time, the pH of supporting electrolyte and MG concentration. Upon optimisation of these parameters, the electrode response was found to be directly proportional to the concentration of MG in the range from 2.06 × 10﹣6 M to 6.74 × 10﹣5 M, leading to a detection limit of 1.84 μM for 240 s preconcentration at ﹣0.1 V. A mechanism was also proposed for the electrochemical oxidation of MG.

Mango malformation: II. mangiferin changes associated with <i>fusarium</i>pathogens

Mangiferin (1,3,6,7-tetrahydroxy xanthone-C2-b-D-glucoside) promoted vegetative growth and exhibited inhibitory role on the occurrence of malformation. Mangiferin changes associated with mango malformation pathogens were followed after inoculated mango seedlings (three years) with malformation pathogens i.e. Fusa-rium subglutinans, F. sterilihyphosum, F. oxysporum and F. proliferatum. Mangiferin remained at lower level in leaves of malformed shoots as compared to healthy one. The floral malformation was observed to be associated with the reduction of mangiferin. Strong positive correlations between mangiferin activity and malformation incidence were observed. Mangiferin level at panicle initiation may give a possible estimate of malformation incidence in mango.

Regulating effects of mangiferin on oxidative stress response and mitochondrial pathway apoptosis in spinal cord injury model

Objective:To study the regulating effects of mangiferin on oxidative stress response and mitochondrial pathway apoptosis in spinal cord injury model.Methods: SD rats were selected as the experimental animals and divided into the control group that underwent sham operation, the injury group that were made into spinal cord injury models and the mangiferin group that were made into spinal cord injury models and given mangiferin intervention. 30 d after the intervention, spinal cord tissue was collected, radioimmunoprecipitation kit was used to determine the contents of oxidative stress indexes ROS, H2O2, SOD, GSH-Px and CAT, and enzyme-linked immunosorbent assay kit was used to determine the contents of apoptosis indexes SKIP, ASK1, Fas, tBid, APAF-1 and Caspase-3.Results:ROS, H2O2, SKIP, ASK1, Fas, tBid, APAF-1 and caspase-3 contents in spinal cord tissue of injury group were significantly higher than those of control group whereas SOD, GSH-Px and CAT contents were significantly lower than those of control group;ROS, H2O2, SKIP, ASK1, Fas, tBid, APAF-1 and caspase-3 contents in spinal cord tissue of mangiferin group were significantly lower than those of injury group whereas SOD, GSH-Px and CAT contents were significantly higher than those of injury group.Conclusion:Mangiferin has inhibitory effect on oxidative stress response and mitochondrial pathway apoptosis in spinal cord injury model.

Mangiferin ameliorates hyperglycemia by inhibiting oxidation and α-glucosidase activity

Objective： Mangiferin （MF） is a polyphenol isolated from the root of Anemarrhena asphodeloides Bge.. This study wasaimed to investigate the effects of MF on hyperglycemia in animal models of insulin resistance and streptozotocin（STZ）-induced diabetes. Methods： The diabetes mellitus model was established in mice by receiving a multiplehypodermic injection of hydrocortisone sodium succinate （HCSS） （70 mg/kg） or a single intravenous injection of STZ（130 mg/kg）. Meanwhile MF at different dosage （50, 100 and 200 mg/kg） were oral administrated for consecutive 10days. Data of blood glucose were collected at different time after intraperitoneal injection of insulin （0.5 U/kg） toinvestigate the insulin resistant. As well as the oxygen radical absorbance capacity （ORAC） and superoxide dismutase（SOD） activity of kidney were measured. The in vitro experiment was established to investigate the inhibitory capacity ofMF to α-glucosidase. Results： Oral administration of MF significantly prevented insulin resistance caused by HCSSinjection. STZ-induced diabetic symptoms were also improved, including fasting blood glucose, glycated hemoglobin,plasma triglycerides, hepatic glycogen, kidney SOD and ORAC level. The in vitro experiment demonstrated that MF hadpotent α-glucosidase inhibitory activity. Conclusion： The obtained results demonstrate that MF ameliorates insulinresistance and STZ-induced glucose metabolism disturbance. The MF exerts the protective effects through improving theantioxidant ability, promoting hepatic glycogen synthesis and inhibiting α-glucosidase activity.

芒果苷对大鼠骨髓间充质干细胞氧化应激损伤的保护作用

背景:间充质干细胞移植治疗脊髓损伤有一定效果,但仍存在局部氧化应激环境导致移植细胞存活率低、细胞移植效率不佳等问题。目的:探究芒果苷在氧化应激状态下对骨髓间充质干细胞的保护作用。方法:取第3代大鼠骨髓间充质干细胞,按浓度梯度0,20,40,80,160μmol/L加入芒果苷孵育2 h,然后加入含400μmol/L H_(2)O_(2)的无血清L-DMEM培养基及相应浓度的芒果苷继续培养12 h及24 h,以常规培养的大鼠骨髓间充质干细胞为正常对照组。MTT法检测各组细胞存活情况,按照试剂盒操作方法检测细胞培养液中超氧化物歧化酶、丙二醛及过氧化氢酶水平。结果与结论:与正常对照组相比,H_(2)O_(2)组细胞的存活能力显著降低(P<0.01),超氧化物歧化酶、过氧化氢酶水平显著降低(P<0.01),丙二醛水平显著升高(P<0.01);与H_(2)O_(2)组相比,芒果苷组细胞的存活能力显著升高(P<0.01),超氧化物歧化酶、过氧化氢酶水平显著升高(P<0.01),丙二醛水平显著降低(P<0.01)。芒果苷在20μmol/L浓度以上表现出浓度依赖性的抗氧化应激保护活性,且在160μmol/L以下无细胞毒性。结果表明,抗氧化剂芒果苷可增加骨髓间充质干细胞的抗氧化活性,为骨髓间充质干细胞移植提供一种新的预处理策略。

Antihyperuricemic effect of mangiferin aglycon derivative J99745 by inhibiting xanthine oxidase activity and urate transporter 1 expression in mice

A mangiferin aglycon derivative J99745 has been identified as a potent xanthine oxidase(XOD) inhibitor by previous in vitro study. This study aimed to evaluate the hypouricemic effects of J99745 in experimental hyperuricemia mice, and explore the underlying mechanisms. Mice were orally administered 600 mg/kg xanthine once daily for 7 days and intraperitoneally injected 250 mg/kg oxonic acid on the 7 th day to induce hyperuricemia. Meanwhile, J99745(3, 10, and 30 mg/kg), allopurinol(20 mg/kg) or benzbromarone(20 mg/kg) were orally administered to mice for 7 days. On the 7 th day,uric acid and creatinine in serum and urine, blood urea nitrogen(BUN), malondialdehyde(MDA) content and XOD activities in serum and liver were determined. Morphological changes in kidney were observed using hematoxylin and eosin(H&E) staining. Hepatic XOD, renal urate transporter 1(URAT1), glucose transporter type 9(GLUT9), organic anion transporter 1(OAT1) and ATP-binding cassette transporter G2(ABCG2) were detected by Western blot and real time polymerase chain reaction(PCR). The results showed that J99745 at doses of 10 and 30 mg/kg significantly reduced serum urate, and enhanced fractional excretion of uric acid(FEUA). H&E staining confirmed that J99745 provided greater nephroprotective effects than allopurinol and benzbromarone. Moreover, serum and hepatic XOD activities and renal URAT1 expression declined in J99745-treated hyperuricemia mice. In consistence with the ability to inhibit XOD, J99745 lowered serum MDA content in hyperuricemia mice. Our resultssuggest that J99745 exerts urate-lowering effect by inhibiting XOD activity and URAT1 expression, thus representing a promising candidate as an anti-hyperuricemia agent.

Effects of ethanol extract of Bombax ceiba leaves and its main constituent mangiferin on diabetic nephropathy in mice

The present study was designed to explore the mechanism by which ethanol extract of Bombax ceiba leaves(BCE) and its main constituent mangiferin(MGF) affect diabetic nephropathy by combating oxidative stress. Oral administration of BCE and MGF to normal and streptozotocin(STZ)-induced diabetic mice were carried out. Fasting blood glucose, 24-h urinary albumin, serum creatinine, and blood urea nitrogen were tested, histopathology, and immunohistochemical analysis of kidney tissues were performed. Moreover, mesangial cells were treated with BCE and MGF for 48 h with or without 25 mmol·L^(-1) of glucose. Immunofluorescence, Western blot and apoptosis analyses were used to investigate their regulation of oxidative stress and mitochondrial function. BCE and MGF ameliorated biochemical parameters and restored STZ-induced renal injury in the model mice. In vitro study showed that high glucose stimulation increased oxidative stress and cell apoptosis in mesangial cells. BCE and MGF limited mitochondrial membrane potential(Δψm) collapse by inhibiting Nox4, mitochondrially bound hexokinase II dissociation, and subsequent ROS production, which effectively reduced oxidative stress, cleaved caspase-3 expression and cell apoptosis. Our work indicated that BCE and MGF had protective effects on diabetic caused kidney injury and prevented oxidative stress in mesangial cells by regulation of hexokinase II binding and Nox4 oxidase signaling.

Mangiferin inhibited neuroinflammation through regulating microglial polarization and suppressing NF-κB,NLRP3 pathway

Inflammation plays important roles in the progress of neurodegenerative diseases,such as Parkinson’s disease and Alzheimer’s disease.Microglia is responsible for the homeostasis of the central nervous system(CNS),and involved in the neuroinflammation.Therefore,it could be potential in treatment of neurodegenerative diseases to suppress the microglia-mediated neuroinflammation.Mangiferin,a major glucoside of xanthone in Anemarrhena Rhizome,has anti-inflammatory,anti-diabetes,and anti-oxidative properties.However,the effect of mangiferin on the inflammatary responses of microglia cells are still poorly understand.In this study,we investigated the mechanism by which mangiferin inhibited inflammation in LPS-induced BV2 microglia cells.BV2 cells were pretreatment with mangiferin followed by LPS stimulation.In vitro assays,NO and cytokines production were quantified.Western blot and immunocytochemistry were used to examine the effect of mangiferin on the polarization of BV2 cells and signaling pathway.The results showed that mangiferin treatment significantly reduced NO,IL-1β,IL-6 and TNF-αproduction,also reduced the mRNA and protein of iNOS and COX-2,promoted the polarization of inflammatory toward anti-inflammatory,and inhibited activation of NF-κB and NLRP3 inflammasome.These data suggest that mangiferin has an anti-neuroinflammatory property via regulating microglia macrophage polarization and suppressing NF-κB and NLRP3 signaling pathway,and may act as a potential natural therapeutic candidate for neuroinflammatory diseases.