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.

Two new saponins from Anemarrhena asphodeloides Bge.

Two new saponins 3-O-β-D-glucopyranosyl (1→2)-β-D-mannopyranosyl sarsasapogenin, named timosaponin A IV(1) and (5β, 25S)-26-O-β-D-glucopyranosyl-furost-20(22)-en-3,26-diol-3-O-β-D-glucopyranosyl (1→4) glucopyranosyl (1→2)-β-D-galacopyranoside, named timosaponin B IV(2), were isolated by silica gel column chromatography and preparative HPLC from Anemarrhena asphodeloides Bge. Their structures were elucidated by chemical characters and spectroscopic analysis.

Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS

A global quality control method based on high performance liquid chromatography(HPLC)coupled with diode array detection(DAD),single quadrupole mass spectrometry(MS)and time-of-flight mass spectrometry(TOFMS)was developed for simultaneous determination of seven major components(mangiferin,neomangiferin,timosaponin E1,timosaponin E,timosaponin BⅡ,timosaponin BⅢ,and timosaponin AⅢ)and identification of most components in extracts of Rhizoma Anemarrhenae(RA).HPLC analysis was performed on an Agilent SB-C18 column(4.6 mm×150 mm,5 μm)by gradient elution using acetonitrile and water-acetic acid(100∶0.05,v/v)as the mobile phase.Seven major components in RA were successfully separated.This quantitative method was fully validated in respect of the following performance criteria:linearity,precision,repeatability,stability,accuracy,limits of detection(LOD)and quantification(LOQ).A formula database of known compounds in RA was established,against which,most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS.This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China.This global quality control method,which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components,is suitable for routine quantification and comprehensive quality control of RA.

Study on the effect of Rhizoma Anemarrhenae on glucose and lipid metabolism in diabetic mice

Objective:To investigate the effect of Rhizoma Anemarrhenae extract on glucose and lipid metabolism in diabetic mice,and provide basis for follow-up research.Methods:The method of streptozotocin combined with high-fat diet was used to replicate the diabetes model in this study.After continuous intragastric administration for 6 weeks,the effects of Rhizoma Anemarrhenae extract on body weight,blood sugar,blood lipids,serum insulin and the organ index in diabetic mice were detected.Results:The fasting blood glucose and blood lipids(serum total cholesterol,triglyceride,low density lipoprotein cholesterol levels)of the Rhizoma Anemarrhenae group of mice were significantly decreased,which was statistically significant compared with the model group.The area under the curve of the Rhizoma Anemarrhenae group is significantly smaller than that of the model group,and the result is statistically significant.Rhizoma Anemarrhenae can increase serum insulin levels in diabetic mice,which is statistically significant compared with the model group.Compared with the model group,the liver,spleen,kidney and pancreas indexes of the Rhizoma Anemarrhenae group are significantly reduced.Conclusion:The extract of Rhizoma Anemarrhenae can significantly reduce blood glucose concentration,lower blood lipid levels,promote serum insulin secretion,and protect liver,kidney,spleen,pancreas and other tissues and organs.

Inhibitory Effects of Saponins From Anemarrhena asphodeloides Bunge on the Growth of Vascular Smooth Muscle Cells

Objective To investigate the effects of saponins from Anemarrhena asphodeloides Bunge (SAaB) (Botanical Name: Anemarrhena Asphodeloidis Rhizoma) on the growth of vascular smooth muscle cells (VSMCs). Methods Cell proliferation was measured by a newly developed cell proliferation reagent, WST-1. Cell apoptosis was assayed by flow cytometry through detecting annexin V. Nitric oxide production was evaluated using confocal laser scanning microscopy with diaminofluorescein diacetate (DAF-2, DA). Cell aldose reductase (AR) activity, as well as the effect of Epalrestat and interleukin-1β were also explored. Results WST assay showed that cell proliferation induced by serum was significantly inhibited by SAaB (P<0.01). Flow cytometry analysis revealed that SAaB could enhance apoptotic rate of VSMCs (P<0.01). Nitric oxide production was significantly enhanced after administration of SAaB and interleukin-1β. Moreover, AR activity of VSMCs was also remarkably inhibited by both SAaB and Epalrestat (P<0.01). Conclusion SAaB can inhibit proliferation and enhance apoptosis of VSMCs. It may protect vascular cells by inhibiting VSMC proliferation and augmenting apoptotic rate of VSMCs via NO-dependent pathway.

基于血清药物化学和网络药理学的知母-黄柏药对盐炙前后治疗2型糖尿病药效物质基础及作用机制分析

目的基于液质联用技术对生知母-生黄柏、盐知母-盐黄柏药对水提液灌胃给药后大鼠入血成分分析,结合网络药理学预测盐炙在知母-黄柏药对治疗2型糖尿病的影响,并通过体外实验进行初步验证。方法大鼠连续灌胃给药生知母-生黄柏药对、盐知母-盐黄柏药对水提液2次,中间间隔1 h,末次给药60 min后腹主动脉取血,用甲醇沉淀蛋白法处理后复溶,采用色谱柱Shim-pack GIST C 18(4.6 mm×150 mm,5μm);流动相A相为0.1%甲酸水,B相为0.1%甲酸-乙腈;梯度洗脱,正、负离子全扫描模式,质量扫描范围m/z 100~1500。结合数据库二级谱图及文献,分析鉴定生知母-生黄柏、盐知母-盐黄柏药对的入血成分。检索2型糖尿病疾病靶点,对入血成分和疾病的交集靶点进行蛋白质互作网络分析、GO、KEGG通路富集分析,构建“入血成分-靶点”网络图,运用AutoDock软件对筛选出的核心成分和核心靶点进行分子对接验证。验证实验以HepG2细胞为实验对象,胰岛素联合高糖诱导胰岛素抵抗模型,CCK8法检验盐炙前后知母-黄柏药对对细胞增殖影响,Western blot检测PI3K-AKT信号通路相关蛋白的表达情况。结果生知母-生黄柏药对水提液大鼠血清中鉴定出15种原型成分,1个芒果苷代谢成分。盐知母-盐黄柏药对水提液大鼠血清中鉴定出17个原型成分,1个芒果苷代谢成分。盐炙后入血成分中芒果苷、小檗碱、3-异丁基戊二酸等成分含量较生品高。KEGG和GO结果显示,知母-黄柏药对治疗2型糖尿病可能与RNA聚合酶的转录调控、炎症反应、AGE-RAGE、PI3K-AKT、胰岛素抵抗等通路有关。细胞实验表明盐炙前后知母-黄柏药对可以上调p-PI3K/PI3K、p-AKT/AKT、GLUT4蛋白表达,且盐炙组效果优于生品组。结论初步阐释了知母-黄柏药对盐炙前后入血成分,阿魏酸、小檗碱、小檗红碱、芒果苷与mTOR、SIRT1、EGFR、PPARA等可能是知母-黄柏药对盐炙后增强2型糖尿病治疗效果的主要成分和靶点,其机制可能是增强了PI3K-AKT等相关通路的作用,为知母-黄柏药对盐炙前后药效物质基础研究及临床应用提供重要参考。

知母皂苷AⅢ对肝癌细胞Akirin2基因表达及细胞增殖的抑制作用

目的:研究Akirin2在人肝癌组织中的表达及知母皂苷AⅢ(TA-Ⅲ)抑制人肝癌细胞(HepG2)增殖和促进凋亡的潜在机制。方法:应用RT-qPCR法和免疫组织化学法检测103例肝癌组织中Akirin2的表达。应用MTT法分别检测Akirin2过表达组、Akirin2 inhibitor组以及不同浓度TA-Ⅲ(0、5、10、20、40 mol/L)、紫杉醇组(80 g/ml)干预后对HepG2细胞增殖活力的影响;平板克隆实验和流式细胞术分别检测Akirin2过表达、Akirin2抑制以及TA-Ⅲ干预后对HepG2细胞克隆、凋亡的影响;Akirin2和凋亡蛋白采用WB法检测。TA-Ⅲ(40 mol/L)处理HepG2细胞的同时转染Akirin2 mimics(TA-Ⅲ+Akirin2过表达组),比较过表达Akirin2对TA-Ⅲ干预细胞增殖、凋亡的影响。结果:肝癌患者肝癌组织中Akirin2呈显著高表达(P<0.05),且Akirin2的高表达与患者的不良预后独立相关(P<0.05)。Akirin2过表达组的细胞增殖能力、克隆能力显著高于对照组(P<0.05),凋亡相关蛋白表达和细胞凋亡率低于对照组(P<0.05)。Akirin2 inhibitor组的细胞增殖能力、克隆能力显著低于对照组(P<0.05),凋亡相关蛋白表达和细胞凋亡率高于对照组(P<0.05)。TA-Ⅲ处理组的HepG2细胞增殖活力显著抑制,细胞增殖活性抑制与干预浓度、时间成正比。TA-Ⅲ不同浓度处理HepG2细胞增殖活力、细胞克隆能力、Akirin2蛋白表达显著低于对照组(P<0.05),细胞凋亡率和凋亡蛋白显著高于对照组(P<0.05)。Akirin2过表达后,TA-Ⅲ对HepG2细胞的细胞增殖能力细胞凋亡率和凋亡相关蛋白的影响作用被部分逆转。结论:Akirin2在肝癌中呈显著高表达,TA-Ⅲ可通过抑制Akirin2表达抑制癌细胞增殖,促进凋亡。

重构本草——知母

通过古籍文献调研、中药现代研究成果的梳理,结合中医临床及中药学等学科领域内专家学者的认识及应用经验,经讨论认为:知母功效主要为清热泻火,滋阴润燥。症靶为发热、口渴、烦扰不宁、肺热燥咳。标靶为血糖升高。现代药理发现知母及其有效成分具有解热、抗炎、抗病原微生物、降糖、改善气道炎症、缓解肺纤维化、镇静催眠、调节交感神经、抗肿瘤、降血脂等作用。本品苦寒滋腻而质滑,凡脾胃虚寒,寒湿内停,大便溏泄者忌服。临床使用剂量为6~90 g,大剂量使用时,须注意避免知母苦寒损伤脾胃,可配伍生姜、干姜、草果、肉桂等辛温药物,并积极探索知母用量与症靶、标靶的量效关系。

人参、知母调控NR2B-CaMKⅡ-AMPAR1通路防治糖尿病认知功能障碍作用研究

目的 观察人参、知母防治糖尿病认知功能障碍(diabetic cognitive impairment,DCI)的作用,并探讨其机制。方法 采用高脂高糖饲料联合腹腔注射链脲佐菌素(STZ)(70 mg/kg)建立DCI小鼠模型,除空白组,造模成功后随机分为模型组、二甲双胍组(30 mg/kg)、多奈哌齐组(30 mg/kg)、人参组(3.6 g/kg)、知母组(10.8 g/kg)、人参知母组(3.6、7.2 g/kg),每组8只。连续灌胃给药30 d,记录各组小鼠体质量和血糖。给药结束用Moriss水迷宫法进行行为学测试,酶联免疫吸附测定(ELISA)法检测小鼠血清中糖化血红蛋白(GHbA1c);苏木素-伊红(HE)染色法观察海马组织病理变化,Western blot法检测NR2B、钙调蛋白激酶Ⅱ(CaMKⅡ)、α-氨基-3-羟基-5-甲基-4-异唑-丙酸谷氨酸受体1(AMPAR1)蛋白的表达。结果 与模型组相比,人参组(3.6 g/kg)、知母组(10.8 g/kg)、人参知母组(3.6、7.2 g/kg)小鼠体质量增长,血糖下降(P<0.05,P<0.01),逃避潜伏期、探索距离显著缩短(P<0.05,P<0.01),站台穿越次数、目标象限游泳时间显著增加(P<0.05,P<0.01),GHbA1c降低(P<0.05,P<0.01),小鼠海马CA1区神经细胞更加整齐紧密,脑组织中NR2B、AMPAR1蛋白表达显著下调(P<0.01),CaMKⅡ蛋白表达显著上调(P<0.01)。结论 人参知母可改善DCI,其机制可能与调控NR2B-CaMKⅡ-AMPAR1通路有关。

知母治疗阿尔茨海默病网络药理学研究及实验验证

目的:探索知母治疗阿尔茨海默病(AD)的作用机制。方法:通过网络药理学筛选知母治疗AD的活性成分、作用靶点,蛋白质-蛋白质相互作用网络构建及核心靶点分析,并进行基因本体(GO)和京都基因和基因组百科全书(KEGG)通路富集分析。提取外周血淋巴细胞并构建淋巴母细胞样细胞系(LCL),建立LCL-SKNMC体外细胞模型。采用MTT法和细胞计数试剂盒8(CCK-8)法分别检测SKNMC细胞和LCL的活性,7´-二氯荧光素二乙酸酯(DCFH-DA)探针检测共培养模型中生成的活性氧,免疫荧光染色检测共培养模型中生成的β淀粉样蛋白1-42(Aβ_(1-42)),蛋白质印迹法检测共培养模型中相关蛋白的表达。分别观察氧化应激、正常状态及热应激状态下N2线虫的寿命,DCFH-DA探针检测N2线虫生成的活性氧,瘫痪实验研究CL4176线虫的瘫痪时间,硫黄素S染色检测CL4176线虫咽部沉积的Aβ。结果:知母具有15个潜在活性成分及103个药物-疾病靶点,可能通过白蛋白、Akt1、肿瘤坏死因子、表皮生长因子受体、血管内皮生长因子A、哺乳动物雷帕霉素靶蛋白、淀粉样前体蛋白(APP)等相关靶点发挥抗AD作用。知母药理作用主要涉及阿尔茨海默病、内分泌抵抗、胰岛素抵抗等生物过程,以及神经活性配体-受体相互作用、磷脂酰肌醇3-激酶(PI3K)-Akt信号通路、钙信号通路、糖尿病并发症中的AGE-RAGE信号通路、神经营养因子信号通路等通路。体外实验显示,知母可减少LCL-SKNMC中活性氧生成和Aβ_(1-42)的产生(均P<0.01),抑制β-分泌酶1、APP、Aβ_(1-42)蛋白的表达(均P<0.05),上调PI3K、Akt、GSK3β蛋白的磷酸化水平(均P<0.05)。体内研究进一步证实,知母可延长应激状态及正常情况下线虫的寿命,减轻活性氧积累及Aβ沉积毒性。结论:知母可减少AD中Aβ的生成,抑制其诱导的氧化应激作用,这些作用可能是通过调控PI3K/Akt/GSK-3β通路实现的。

中药知母组培快繁体系的建立

为了建立知母高效组织培养及快速繁殖体系,将知母的叶片、根茎和幼嫩花葶分别接种于9种组合培养基上,筛选最佳的外植体材料;以MS培养基为基本培养基,通过不同种类激素和浓度的配比试验,筛选出知母离体组培快繁不同阶段的培养基组合;测定不同种类激素和浓度组合培养基下炼苗移栽成功的知母组培苗芒果苷含量。结果表明,幼嫩花葶为诱导愈伤组织最佳外植体,诱导知母幼嫩花葶产生愈伤组织的培养基组合为MS+1.0 mg/L KT+0.5 mg/L NAA,出愈率为79.40%;诱导愈伤组织产生不定芽的培养基组合为MS+0.5 mg/L 6-BA+0.5 mg/L NAA,生芽率为100%;诱导不定芽生根培养的培养基组合为MS+0.5 mg/L NAA,生根率为100%,且在此培养基组合下炼苗移栽的知母组培苗芒果苷含量最高。

Screening potential α-glucosidase inhibitors from Anemarrhena asphodeloides using response surface methodology coupled with grey relational analysis

Objective: To screen potential α-glucosidase inhibitors from Anemarrhena asphodeloides.Methods: Response surface methodology employing Box-Behnken design was used to optimize conditions for the extraction of α-glucosidase inhibitory active compounds from A. asphodeloides. The powders(20.0 g) of A. asphodeloides were extracted under the optimized conditions. The extract was applied to a D-101 macroporous resin column. It was eluted with ethanol and water to give six fractions. Compounds from the active fraction were identified by UPLC-Q-TOF-MS. The structure-activity relationship was discussed based on grey relational analysis.Results: The optimum extraction conditions were as follows: ethanol concentration, 100%; extraction temperature, 51 ℃; and solvent to solid ratio, 23 mL/g. It indicated that the active compounds were concentrated into 80% ethanol fraction. Twenty five steroid saponins from 80%ethanol fraction were identified by UPLC-Q-TOF-MS. Peaks 19 and 23 were tentatively identified as new structures. The predicted α-glucosidase inhibitory activities of the compounds were 7 > 2 > 1 > 22 > 23 > 3 > 9 > 21 > 24 > 4 > 13 > 8 > 14 > 16 > 17 > 25 > 6 > 19.Conclusion: The fraction eluted by 80% ethanol showed the best inhibitory activity. After analyzing the data of UPLC-Q-TOF-MS, 25 steroid saponins were tentatively identified in this fraction.

Two new steroidal saponins isolated from Anemarrhena asphodeloides

Two new steroidal saponins,named timosaponin P(1) and timosaponin Q(2),were isolated from the rhizome parts of Anemarrhena asphodeloides Bunge using various chromatographic methods.Their structures and absolute configurations were elucidated by a combination of spectroscopic and spectrometric data,including 1D,2D NMR,HR-ESI-MS and ECD calculations,and this is the first time the absolute configuration of C-23 of steroidal saponin was confirmed by ECD calculations.

Exploring the catalytic function and active sites of a novel C-glycosyltransferase from Anemarrhena asphodeloides

Anemarrhena asphodeloides is an immensely popular medicinal herb in China,which contains an abundant of mangiferin.As an important bioactive xanthone C-glycoside,mangiferin possesses a variety of pharmacological activities and is derived from the cyclization reaction of a benzophenone C-glycoside(maclurin).Biosyntheti-cally,C-glycosyltransferases are critical for the formation of benzophenone C-glycosides.However,the benzo-phenone C-glycosyltransferases from Anemarrhena asphodeloides have not been discovered.Herein,a promiscuous C-glycosyltransferase(AaCGT)was identified from Anemarrhena asphodeloides.It was able to catalyze efficiently mono-C-glycosylation of benzophenone,together with di-C-glycosylation of dihydrochalcone.It also exhibited the weak O-glycosylation or potent S-glycosylation capacities toward 12 other types of flavonoid scaffolds and a simple aromatic compound with–SH group.Homology modeling and mutagenesis experiments revealed that the glycosylation reaction of AaCGT was initiated by the conserved residue H23 as the catalytic base.Three critical residues H356,W359 and D380 were involved in the recognition of sugar donor through hydrogen-bonding interactions.In particular,the double mutant of F94W/L378M led to an unexpected enzy-matic conversion of mono-C-to di-C-glycosylation.This study highlights the important value of AaCGT as a potential biocatalyst for efficiently synthesizing high-value C-glycosides.

桂枝汤类方在风湿痹病中的应用

风湿痹内因为正气不足、机体营卫失调,外因为感受风寒湿等外邪,外湿侵袭肌表,正气不足,营卫失调,腠理疏松,汗出当风,外邪入体,流窜全身、流注关节,发为风湿痹病,治应调和营卫。桂枝汤用于治疗风寒邪伤人肌表,腠理疏松,卫外不固,营阴不守之证,切中风湿痹病病机。风重于湿,痹着肌表,阻滞营卫,气血不利,选桂枝附子汤;太阳中风兼经气不利者,选桂枝加葛根汤;风湿侵袭入体,在体郁遏日久,化热伤阴者,选桂枝芍药知母汤;营卫气血俱不足之血痹重证,选黄芪桂枝五物汤。

滋肾通关方中肉桂反佐配伍对黄柏-知母药对在大鼠体内活性成分组织分布的影响

目的探讨滋肾通关方中肉桂反佐配伍对黄柏-知母药对的活性成分在正常大鼠体内组织分布的影响及其作用机制。方法将56只Wistar大鼠随机分为空白组(等量生理盐水),滋肾通关方低、中、高剂量组(5,10,20 g/kg),黄柏-知母药对低、中、高剂量组(5,10,20 g/kg),各8只。分别灌胃相应药物1 h后取大鼠心、肝、脾、肺、肾组织并制备匀浆液,采用超高效液相色谱串联质谱法测定大鼠体内组织匀浆液中黄柏碱和知母皂苷AⅢ的含量。结果与黄柏-知母药对低、中、高剂量组比较,滋肾通关方低、中、高剂量组大鼠心、肝、脾、肺、肾组织匀浆液中黄柏碱和知母皂苷AⅢ的含量整体更高;滋肾通关方高剂量组大鼠肺组织匀浆液中黄柏碱和知母皂苷AⅢ的含量显著高于黄柏-知母药对高剂量组(P<0.05),滋肾通关方低剂量组大鼠心组织中黄柏碱的含量显著高于黄柏-知母药对低剂量组(P<0.05)。结论滋肾通关方中肉桂反佐配伍黄柏-知母药对的作用机制可能与改变药对中活性成分在组织中的分布有关。

知母标准汤剂的质量评价方法及指标成分的量值传递规律探索

目的:建立知母标准汤剂的质量评价方法,明确10批知母标准汤剂中指标成分的含量及转移率范围,探索从饮片到标准汤剂,指标成分的量值传递规律。方法:制备10批知母标准汤剂,采用超高效液相色谱-二极管阵列检测器法(UPLC-PDA)对指标成分芒果苷进行含量测定,以0.1%甲酸水(A)-乙腈(B)为流动相进行梯度洗脱(0~3 min,10%~13%B;3~6 min,13%~14%B;6~7 min,14%~90%B),检测波长为258 nm;采用高效液相色谱-蒸发光散射检测器法(HPLC-ELSD)对知母皂苷BⅡ进行含量测定,以乙腈-水(25∶75)为流动相进行等度洗脱,载气流速为2.8 L·min^(–1),漂移管温度为104℃。结果:建立的指标成分含量测定方法的方法学验证均良好,能用于知母标准汤剂中2个指标性成分的定量检测。10批知母标准汤剂中芒果苷、知母皂苷BⅡ的质量分数分别为0.88%~1.12%和0.88%~2.53%,芒果苷含量位于其均值的±30%范围内;转移率分别为45.83%~57.27%和26.94%~43.49%,均在其均值的±30%范围内。结论:建立了知母标准汤剂的质量评价方法,指标成分芒果苷和知母皂苷BⅡ在不同批次的知母饮片与标准汤剂间的转移率一致性较好,表明这2个指标成分在标准汤剂的制备过程中转移情况稳定,为源于知母水煎液的制剂的质量控制方法的制定提供参考。

知母黄芪粗提物对蛋鸡产蛋性能、蛋品质及抗氧化能力的影响

试验旨在研究在蛋鸡饮水中添加知母黄芪粗提物对蛋鸡产蛋性能、蛋品质及抗氧化功能的影响。选择体重相近的1200只伊莎褐蛋鸡,随机分成4个处理组,各组蛋鸡饲喂相同的日粮,各试验组分别在饮水中添加1.0、1.5、2.0 mL/kg的知母黄芪粗提物,对照组饮水中不添加知母黄芪粗提物。预试期1 w,正式试验期4 w。结果显示,与对照组相比,试验2组和试验3组伊莎褐蛋鸡的产蛋率、平均蛋重显著提高(P<0.05);所有知母黄芪粗提物组的蛋鸡的死淘率显著降低(P<0.05);试验2组和试验3组伊莎褐蛋鸡的料蛋比、鸡蛋破损率显著降低(P<0.05)。试验2组和试验3组鸡蛋的哈夫单位显著提高(P<0.05);所有知母黄芪粗提物组鸡蛋的蛋壳强度和厚度得到显著改善(P<0.05);试验2组和试验3组鸡蛋的蛋重和蛋清重显著提高(P<0.05)。与对照组相比,知母黄芪粗提物组伊莎褐蛋鸡血清中丙二醛水平显著下降(P<0.05),试验2组和试验3组伊莎褐蛋鸡血清总抗氧化能力显著增强(P<0.05),过氧化氢酶、超氧化物歧化酶及谷胱甘肽过氧化物酶活性显著增强(P<0.05),而过氧化氢浓度显著降低(P<0.05)。研究表明,在蛋鸡养殖饮水中添加1.5、2.0 mL/kg的知母黄芪粗提物可以改善蛋鸡的产蛋性能、蛋品质,增强机体的抗氧化能力,建议在养殖中使用1.5 mL/kg的知母黄芪粗提物。

清热药知母及其配伍防治糖尿病肾病的效应机制分析

糖尿病肾病(diabetic nephropathy,DN)发病机制较为复杂,与炎症反应、氧化应激损伤、肾脏血流异常、糖脂代谢紊乱等因素有关,且各机制间相互影响。现代医学多采用控制糖脂及血压等方式治疗DN,患者需长期服药、经济负担重且不良反应大,但无法从根本上避免DN进展。而中医药防治DN具有多层次、多环节、多途径的复合效应,可标本兼顾,呈现出降脂、降黏、抗炎及抗氧化等药理作用,能延缓DN进展,减轻患者经济负担并提高其生活质量。DN病始阴虚燥热,中期热耗精津,后期瘀水并见,热邪贯穿疾病始终,中医多采用清热养阴兼活血的治法防治DN进展,疗效确切。知母性寒、味甘苦,擅于清热泻火、滋阴润燥,临床用于防治DN及糖尿病(diabetes mellitus,DM),含有芒果苷、知母皂苷和知母多糖等多种化学成分,常与石膏、黄柏及葛根相须配伍,也用于白虎汤、知柏地黄丸等经典复方,可显著降低DN患者糖脂及尿蛋白、减轻其临床症状。知母的单体-芒果苷,能有效减轻炎症反应及氧化应激损伤,临床常与石膏相须配伍,但此药对防治DN的文献较少。因此,还需研究更多中药单体、药对及复方防治DN的机制。基于清法学说及DN病因病机,从分子生物学角度对知母及其配伍防治DN的机制作一综述,以期为中医药防治DN提供理论支撑。

知母-黄柏药对盐炙前后HPLC指纹图谱及多指标成分含量变化

目的建立知母-黄柏药对盐炙前后HPLC指纹图谱,对11种指标性成分进行定量分析,探索药对盐炙前后的质量差异。方法采用HPLC法建立知母-黄柏药对盐炙前后指纹图谱,通过化学计量学方法对知母-黄柏药对盐炙前后指纹图谱进行分析;并采用HPLC法测定知母-黄柏盐炙前后药对中新芒果苷、绿原酸、隐绿原酸、黄柏碱、芒果苷、木兰花碱、药根碱、小檗红碱、小檗碱、柠檬苦素、黄柏酮的含量。结果10批知母-黄柏药对盐炙前后HPLC指纹图谱组内相似度均>0.988,聚类分析、偏最小二乘法-判别分析可明显将生知母-生黄柏、盐知母-盐黄柏分为2类,小檗碱、新芒果苷、芒果苷、小檗红碱、黄柏碱等成分可能是引起两者质量差异的主要标志成分。定量分析显示,知母-黄柏药对盐炙后芒果苷、黄柏碱、药根碱、小檗红碱、小檗碱的含量增加,而新芒果苷、绿原酸、隐绿原酸的含量降低,柠檬苦素的含量亦有降低趋势,木兰花碱、黄柏酮的含量无明显变化。结论所建立的指纹图谱及多指标含量测定方法稳定可靠,从定性与定量评价了知母-黄柏药对盐炙前后的质量差异;为知母-黄柏药对药味炮制研究提供一定的依据。