Optimization of Extraction Process and Content Determination of Total Flavonoids from Sanguisorbae Radix and Carbonized Sanguisorba Root by Orthogonal Experimental Design

[Objectives] To optimize the extraction process of total flavonoids from Sanguisorbae Radix and carbonized Sanguisorba root,compare quality of different batches of Sanguisorbae Radix,study the effects of processing on the content of flavonoids,and provide scientific basis for reasonable utilization of Sanguisorbae Radix. [Methods] Test samples were prepared by heating,refluxing,and extraction,the extraction process was optimized by orthogonal experiment design,color was developed by NaNO_2-Al( NO_3)3-NaOH,and total flavonoids were measured by UV method at the wavelength of 510 nm. [Results] The linear relationship of rutin was excellent in the concentration range of 0. 1248 mg/mL-0. 5712 mg/mL,R^2= 0. 9997; the average recovery was 99. 67% and the RSD was 0. 70%. The optimum extraction conditions were as follows: the volume fraction of ethanol was 50%,the extraction temperature was 90℃,the extraction time was 90 min,and the solid-to-liquid ratio was 1∶ 20( g/mL). [Conclusions] After optimization of the extraction process,the extraction rate of total flavonoids in samples of Sanguisorbae Radix was significantly increased; there was certain difference in the content of total flavonoids between different batches of Sanguisorbae Radix and processed products; the total flavonoids significantly declines in carbonized sanguisorba root,and the influence of processing on its curative effect was to be further studied.

A NEW TRITERPENOID FROM SANGUISORBA ALPINA

From the roots of Sanguisorba alpina, a new triterpenoid was isolated and identified as 2-isobutyryloxy-19α, 25-dihydroxyursa-1, 12-dien-28-oic acid by means of spectral methods, named as aipinic acid.

Anti-tumor activity of Sanguisorba officinalis L.in non-small cell lung cancer and induced apoptosis via PI3K/Akt/mTOR signaling pathway

OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L.(SOL)in non-small cell lung cancer(NSCLC)in vitro and in vivo based on network pharmacology.METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets.Cell counting kit 8(CCK-8)and 5-ethynyl-2′-deoxyuridine(EdU)staining,Western blotting,flow cytometry of AnnexinⅤ/PI,Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro.H&E staining,immunohistochemistry staining and Western blotting were performed in vivo.RESULTS Based on network prediction,we analyzed the 208 common targets of SOL and NSCLC.36 core targets in 208 common targets were obtained through cytoscape analysis.And the top 10 core targets included Akt,mTOR,EGFR,etc..KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway.Furthermore,the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo.The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay.Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo.SOL also significantly inhibited the migration and invasion of A549 and H1299 cells.SOL significantly increased the percentage of cells with PI signal in A549 and H1299,and the process of cell death of A549 cells indicated that SOL induced apoptosis.The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner.The results in vivo were consistent with those in vitro.CONCLUSION SOL-induced,caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC,which further clarified the mechanism of SOL in the inhibition of NSCLC,and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.

Study on the Determination Method for the Content of Tannin in Sanguisorba officinalis L.

[Objectives] To establish a method for the determination of tannin in Sanguisorba officinalis L.,and provide guarantee for further control over quality of semi-finished tannin products from Sanguisorba officinalis L. [Methods] With gallic acid as reference substance,the UV method was used to determine the content of tannin in Sanguisorba officinalis L.,and the methodological examination was done. [Results]With gallic acid as standard,when the concentration of tannin in Sanguisorba officinalis L. was 1. 02-10. 14 μg·m L^(-1),there was a good linear relationship with absorbance,the precision,reproducibility and recovery were in line with the requirements,and the absorbance value was stable within 40 min after coloration. [Conclusions] The method was simple,rapid and accurate in determining the content of tannin in Sanguisorba officinalis L.

近五年地榆的药理作用机制研究进展

传统中药地榆具有凉血止血、解毒敛疮的功效。现代研究对地榆的止血凝血作用、抗氧化作用、抗过敏作用、抗炎作用、抗菌作用、抗肿瘤作用、肾保护作用、升高白细胞作用等药理机制进行了广泛研究,机制明确。其中止血凝血作用机制是通过作用凝血酶及促进巨核细胞分化和血小板生成;抗氧化作用机制是上调抗氧化酶表达和清除氧自由基;抗过敏作用机制是抑制β-己糖氨酶和白介素-4的产生、释放或促进血红素加氧酶-1表达及激活;抗炎作用机制与巨噬细胞极化、聚集和迁移、脂质代谢、改善肠道菌群失调、信号传导与转录激活因子1及细胞外调节蛋白激酶激活、自噬、核因子κB/含pyrin结构域NOD样受体家族3、Toll样受体4/核因子κB、磷酯酰肌醇-3-激酶—丝氨酸/苏氨酸激酶/核因子κB/信号转导与转录激活因子3信号通路等相关;抗菌作用机制破坏细菌细胞壁、细胞膜结构和内部微观结构、干扰细菌总蛋白的合成、降低细胞膜流动性及改变代谢通路等相关基因表达,或抑制青霉素结合蛋白2a表达和mecA基因表达;抗肿瘤作用机制与血管内皮生长因子/磷酯酰肌醇-3-激酶—丝氨酸/苏氨酸激酶/雷帕霉素靶蛋白、表皮生长因子受体/丝裂原活化蛋白激酶表皮生长因子受体/磷酯酰肌醇-3-激酶—丝氨酸/苏氨酸激酶/核因子κB、缺氧诱导因子1、丝裂原化蛋白激酶、丝裂原活化蛋白激酶、肾素—血管紧张素/丝裂原活化的细胞外信号调节激酶/细胞外调节蛋白激酶、Wnt、自噬等信号通路及脂质磷酸酶活性相关;肾保护作用机制是抗氧化、抗炎、抗脂质过氧化、抑制肾小管上皮细胞的增殖;升高白细胞作用机制与自噬、造血系统相关通路和相关基因、造血生长因子等表达、糖代谢、氨基酸和核苷酸代谢的途径相关。在临床上用于消化道出血等出血症、过敏性紫癜及急性湿疹、烧烫伤、痤疮及压疮、急慢性肾小球肾炎等,且地榆制剂地榆升白片常用于白细胞减少症,临床效果明确,表明应用现代科学技术促进了传统中药地榆临床应用。本文对地榆的药理作用机制研究进行综述,为地榆的研究及临床应用提供参考。

3种地榆属植物叶片和茎结构的比较研究

为充分揭示不同地榆属(Sanguisorba)植物对环境的适应性,以地榆(S.officinalis)、细叶地榆(S.tenuifolia)和宽蕊地榆(S.applanata)的叶片和茎为材料,采用常规石蜡切片技术对其进行解剖学比较研究。结果表明,3种地榆属植物叶片形态表现出同一性和差异性,其显微结构呈现典型的双子叶植物特征,上表皮细胞均大于下表皮细胞;细叶地榆、宽蕊地榆与地榆相比在叶脉厚度、栅栏组织厚度上差异显著,在维管束长度、上下角质层厚度、下表皮细胞厚度上无显著差异;茎解剖结构显示地榆维管束数量多于细叶地榆与宽蕊地榆,维管束面积表现为宽蕊地榆>细叶地榆>地榆。上述研究明确了地榆属植物的生物学特性,可为地榆属植物的栽培和高效开发利用提供理论依据。

Chemical Constituents in Charred Sanguisorbae Radix

Objective To study the chemical constituents in the effective fractions of charred Sanguisorbae Radix. Methods The compounds were isolated and purified by column chromatography and their structures were identified on the basis of physicochemical properties and spectral analysis. Results Five compounds were isolated and identified as 3β-hydroxy-28-norurs-17,19,21-trien (1), 3β-hydroxy-28-norurs-12,17-dien (2), 3β,19α-dihydroxyurs-13(18)- en-28-oic acid (3), 3β-[(α-L-arabin-opyranosyl) oxy]-28-norurs-12,17-dien (4), and pomolic acid (5). Conclusion Compounds 1, 3, and 4 are novel compounds belong to triterpenoids and triterpenoid saponins, named as sanguisorbigenins Z, Y1 , and Y2 , respectively.

地榆通过影响PPARG和SLC7A11/GPX4表达减轻溃疡性结肠炎小鼠损伤

目的基于生物信息学探究地榆(Sanguisorbae Radix,SR)改善小鼠溃疡性结肠炎的机制。方法利用R语言对高通量基因表达数据库(GEO)中GSE92415数据集进行溃疡性结肠炎(UC)差异表达基因筛选和加权基因共表达网络分析(WGCNA),并结合FerrDb数据库,获得UC相关铁死亡特征基因。对特征基因进行蛋白互作分析(PPI)和相关性分析,筛选UC铁死亡核心基因。构建葡聚糖硫酸钠(DSS)诱导的UC小鼠模型,并灌胃给予SR水提物9 d,记录疾病活动指数(DAI)和结肠长度,采用HE染色法观察结肠组织病理变化,酶联免疫吸附测定法(ELISA)检测小鼠结肠组织炎症因子肿瘤坏死因子(TNF)⁃α、白细胞介素-6(IL⁃6),生化试剂盒检测脂质过氧化因子丙二醛(MDA)、谷胱甘肽(GSH)水平,免疫荧光法检测结肠组织闭锁小带蛋白1(ZO⁃1)表达,蛋白免疫印迹法(Western blot)检测结肠组织过氧化物酶体增殖物激活受体γ(PPARG)、溶质载体家族7成员11(SLC7A11)和谷胱甘肽过氧化酶4(GPX4)蛋白表达。结果通过生物信息学筛选得到9个UC相关铁死亡特征基因,其中核心基因为PPARG。相关性分析发现PPARG与铁死亡高度相关。结合生物信息学筛选结果,探讨SR改善小鼠UC的机制,实验结果发现,SR可降低UC小鼠DAI值,缓解结肠缩短,改善肠道粘膜屏障功能,对TNF⁃α、IL⁃6、MDA、GSH水平有显著回调作用,并可提高结肠组织PPARG、SLC7A11和GPX4蛋白表达。结论铁死亡与UC密切相关,SR可通过影响PPARG和SCL7A11/GPX4蛋白,进而改善UC小鼠结肠上皮损伤和功能障碍,为UC治疗策略提供了思路与方向。

Protective Effects of Tannins in Sanguisorbae Radix on Myelosuppression Mice

Objective To explore the protective effects of tannins in Sanguisorba Radix （TSR） on myelosuppression mice induced by cyclophosphamide （CTX）. Methods TSR was ig given at the dose of 20 mg/kg for 10 d after ip administration of CTX （200 mg/kg）. Results TSR could significantly increase the numbers of white blood ceils, red blood cells, and platelets of myelosuppression in mice. And it could accelerate bone marrow haemopoietic stem/progenitor cells （HSPCs） in myelosuppression mice and enhance cell proliferation by promoting cell cycles from G0/G1 phase to access into S and G2/M phases, then the reduced number of HSPCs induced by CTX was reversed. Moreover, TSR could increase the mRNA and protein expression levels of O（6）-methylguanine- DNA methyltransferase （MGMT） in HSPCs of myelosuppression mice. Concision TSR has a protective function against CTX-induced myelosuppression. The mechanism might be related to protecting hematopoietic stem cells of bone marrow, stimulating hematopoiesis recovery, as well as preventing the apoptosis of hematopoietic stem cells induced by CTX.

Two new phenylpropanoid glycosides from Sanguisorba officinalis

Two new phenylpropanoid glycosides, 9-O-[6-O-acetyl-β-D-glucopyranosyl]-4-hydroxycinnamic acid （1） and 8-O-β-D-glucopyranosyl-（R）-（＋）-3,4,8-trihydroxy methyl phenylpropionate （2） were isolated from the 80% EtOH extract of the roots of Sanguisorba officinalis. Their slructures were characterized by spectroscopic analysis and chemical method, including 1D NMR, 2D NMR, and HR-ESI-MS. Compounds 1 and 2 exhibited the moderate antimicrobial activities against all Gram-positive and Gram-negative bacteria tested.

Anti-inflammatory Effects of Sanguisorbae Radix on Contact Dermatitis Induced by Dinitrofluorobenzene in Mice

Objective:To investigate the anti-inflammatory effects of Sanguisorbae Radix on contact dermatitis(CD).Methods:Mice were sensitized by painting 30μL of 1-fluoro-2,4-dinitrofluorobenzene(DNFB)onto each ear for 3 days.Four days later,mice were challenged by painting with 50μL of DNFB onto the shaved dorsum every 2 days.Sanguisorbae Radix methanol extract(MESR)was applied onto the shaved dorsum every 2 days.The effects of MESR on skin thickness,skin weights,histopathological changes,skin lesions and cytokine production in DNFB-induced CD mice were investigated,as well as its effects on body weights and spleen/body weight ratio.Results:Topical application of MESR effectively inhibited enlargement of skin thickness and weight(P<0.05).MESR treatment also inhibited hyperplasia,spongiosis and immune cell infiltration induced by DNFB in inflamed tissues and improved lesions on dorsum skin in CD mice.Moreover,treatment with MESR suppressed the increase in the levels of tumor necrosis factor α(TNF-α,P<0.01)and interferonγ(IFN-γ,P<0.05),respectively.Finally,MESR had no effect on body weight gain or spleen/body weight ratio.Conclusion:These data suggest that MESR acts as an anti-inflammatory agent that decreases the production of TNF-α and IFN-γ,resulting in reductions of skin lesions and histopathological changes in inflamed skin tissues.

Two New Triterpenoids from the Roots of Sanguisorba officinalis L.

Two new triterpenoids, octanordammar- 1,11,13(17)-trien- 17-ol-3,16-dione (1) and lup- 12-en- 15α,19β-diol-3,11-dioxo-28-oic acid (4), as well as 13 known compounds were isolated from the roots of Sanguisorba officinalis L. (Rosaceae). Their structures were determined using spectroscopic methods.

地榆乙醇提取物抑制JAK2/STAT3信号通路改善溃疡性结肠炎

目的:本研究旨在探究地榆乙醇提取物的主要化学成分及其治疗溃疡性结肠炎的活性评价及作用机制。方法:通过高分辨液相质谱联用技术鉴别地榆乙醇提取物中主要化学成分;ICR小鼠随机分为空白对照组、DSS模型组、地榆乙醇提取物高、中、低剂量组和美沙拉嗪阳性药组,3%DSS水溶液诱导溃疡性结肠炎小鼠模型,每天观察和记录小鼠的体重、粪便状态和便血情况。组织切片H&E、PAS染色观察各组小鼠结肠组织病理学变化。RT-qPCR和Western blot探究地榆乙醇提取物治疗溃疡性结肠炎的作用机制。结果:高分辨液相质谱联用鉴别出地榆乙醇提取物中鞣质、酚酸、黄酮、黄酮苷、萜类及其皂苷衍生物等共49种化合物成分。地榆乙醇提取物有效改善结肠萎缩和临床症状;H&E和PAS染色表明其能减轻结肠组织炎症细胞的浸润,黏膜上皮细胞的坏死脱落,肠腺隐窝结构及肠黏液层屏障的破坏;显著降低M1巨噬细胞相关的促炎细胞因子mRNA转录水平(IL-1β、IL-6、iNOS和TNF-α),并促进M2巨噬细胞相关的抗炎细胞因子mRNA转录(IL-10、TGF-β和ARG-1);地榆乙醇提取物有效下调JAK2/STAT3信号通路上IL-1β、iNOS、JAK2和P-STAT3关键蛋白的表达水平。结论:本研究基于高分辨液相质谱联用技术,鉴定出地榆乙醇提取物中49种主要化学成分。地榆提取物可通过调节结肠组织M1/M2型巨噬细胞相关细胞因子的平衡和抑制JAK2/STAT3信号通路,发挥保护作用。地榆乙醇提取物未来有望成为治疗UC的有效药物。

炭炒地榆鞣质提取物的壳聚糖PLGA乳液抑菌活性研究

为了探讨炭炒地榆鞣质提取物的壳聚糖PLGA(SCCP)乳液对金黄色葡萄球菌(Staphylococcus aureus,S.aureus)的杀伤作用,用96孔板微量肉汤稀释法测定其对S.aureus的最低抑菌浓度、最小杀菌浓度和联合药敏试验检测抑菌活性。结果显示,与炭炒地榆鞣质提取物相比,SCCP的抑菌活性更强,对S.aureus的最小杀菌浓度为0.063 mg/mL。表明SCCP对S.aureus有较好的抑菌活性,有望开发成新型中药抗菌制剂。

苗药川乌配伍地榆前后新乌头碱、乌头碱、次乌头碱的含量研究

目的研究川乌-地榆冻干粉(1∶2)酒溶液、水溶液中新乌头碱、乌头碱、次乌头碱的含量变化。方法采用高效液相色谱法(HPLC)对川乌配伍地榆前后新乌头碱、乌头碱、次乌头碱的含量进行测定。结果川乌-地榆冻干粉(1∶2)酒溶液3种双酯型生物碱的含量较单川乌冻干粉酒溶液下降了97.3%;川乌-地榆冻干粉(1∶2)水溶液3种双酯型生物碱的含量较单川乌冻干粉水溶液下降了96.7%。结论川乌-地榆冻干粉(1∶2)酒溶液、水溶液可降低川乌中新乌头碱、乌头碱、次乌头碱的含量,起到减毒的作用,为揭示制川乌减毒配伍机制提供指导。

中药治疗幽门螺旋杆菌感染的研究进展

幽门螺旋杆菌(Hp)是一种革兰氏阴性菌,主要定植于胃和十二指肠,可引起胃炎、消化性溃疡和胃癌等疾病。Hp相关的胃癌是人类最常见的恶性肿瘤之一,是全球癌症相关死亡的第四大原因,严重威胁着人类的健康,因此Hp的根除显得尤为重要。但随着抗生素的滥用,该菌的耐药性也大大提升,以往的研究方案逐渐显露出弊端,而中药治疗具有复发率低、不良反应少、不易产生耐药性等独特优势。充分发挥中药优势,不断研究新的治疗方案对Hp的根除具有重大意义。该文主要对Hp的病理机制和治疗进展、中药治疗进行综述。

DISTRIBUZIONE DI Sanguisorba dodecandra MORETTI IN VAL DI SCALVE

L’areale di Sanguisorba dodecandra Moretti,endemismo orobico,mentre risultasufficientemente definito per il versante valtellinese,all’opposto,per il versante berga-masco è stato finora studiato e delineato solo in modo frammentario (FORNACIARI,1948; PIROLA,1964; BECHERER,1970; FUCHS-ECKERT,1990).Si è ritenuto

放线菌、地榆和大蒜防治烟草青枯病试验

选取贵州省遵义市务川仡佬族苗族自治县茅天镇红星村烟田,以放线菌F11菌液、地榆中药液和大蒜粗提液作为防治烟草青枯病的生物药剂,农用链霉素作为药剂对照,清水作为空白对照,开展田间试验,检测实际防治效果。结果表明,平均相对株防效表现为F11菌液(67.96%)>大蒜粗提液(57.48%)>地榆中药液(43.32%)>农用链霉素(28.16%)。

基于网络药理学的苦参-地榆治疗溃疡性结肠炎、肛门湿疹的“异病同治”机制分析

目的:预测苦参-地榆药对治疗溃疡性结肠炎(UC)和肛门湿疹“异病同治”的作用机制。方法:采用网络药理学方法,基于中药系统药理学分析平台(TCMSP)及检索相关文献,寻找苦参-地榆药对中活性成分及作用靶点,并以口服利用度(OB)≥30%,类药性(DL)≥0.18为条件进行筛选;利用OMIM、TTD、Drug Bank数据库检索UC和肛门湿疹疾病相关靶点;通过R语言将苦参-地榆药对与UC和肛门湿疹靶点做Venn图;采用Cytoscape软件绘制疾病-药对-成分-靶点网络;利用STRING数据库,构建蛋白互作网络模型(PPI),并通过Cytoscape软件进行优化;基于Matescape数据平台,对药对治疗UC和肛门湿疹的共同靶点进行基因本体(GO)生物功能和基因组百科全书(KEGG)通路富集分析。结果:筛选后得出,苦参-地榆药对活性成分57个、靶点179个,UC靶点561个,肛门湿疹靶点677个,药对与两疾病交集靶点49个,包括IL-6、JUN、TNF、AKT1等。GO生物过程分析主要涉及对脂多糖的反应、对有毒物质的反应、对活性氧的反应等生物过程;KEGG富集分析共得到15条信号通路,主要包括IL-17信号通路、癌症通路、细胞因子受体相互作用等。结论:苦参-地榆药对“异病同治”UC和肛门湿疹是通过其多成分、多靶点、多通路的特点实现的,为进一步临床拓展应用及实验验证提供参考依据。

不同炮制时间地榆炭化学成分与粉末色度值的相关性分析

目的分析地榆炭炮制过程中饮片粉末色度值与化学成分的关联性。方法采用高效液相色谱法,建立地榆及不同炮制时间地榆炭饮片的特征图谱,采用分光测色仪测定地榆及不同炮制时间地榆炭饮片粉末的色度值[明暗度值(L^(*))、红绿色度值(a^(*))、黄蓝色度值(b^(*))、总色度值(E^(*)ab)],分析外观色泽指标与特征图谱的相关性。结果随着炮制时间的延长,地榆炭较地榆饮片特征图谱的相似度呈降低趋势;L^(*)、a^(*)、b^(*)、E^(*)ab呈减小趋势;HPLC特征图谱13个特征峰中峰5~10、12(阿魏酸)7个色谱峰单位峰面积逐渐减小,均与色度值L^(*)、b^(*)、E^(*)ab成显著正相关;峰2、3、11、13(鞣花酸)单位峰面积先增大后减小,除峰2外,单位峰面积均与色度值L^(*)、b^(*)、E^(*)ab成显著负相关;逐步回归分析结果显示,可通过色度值预测峰3、5~10、12(阿魏酸)、13(鞣花酸)9个色谱峰单位峰面积大小;聚类分析将16个样品分为2类;正交偏最小二乘法判别分析筛选出3个VIP值大于1的差异性成分。结论地榆炭饮片粉末的色度值与其化学成分的含量变化具有相关性,可作为地榆炭炮制过程中监测饮片质量的依据,以提高地榆炭饮片的质量水平及一致性,建议地榆炭饮片炮制终点为6~8 min(260~280℃)。