Activation of GSDME by Lithospermum erythrorhizon drives pyroptotic cell death

Objective:To reveal GSDME-executed pyroptosis in cancer cells induced by the Chinese traditional herbal medicine plant Lithospermum erythrorhizon(L.erythrorhizon,Zi Cao)and to investigate the potential mechanism.Methods:L.erythrorhizon was extracted by ultrasonication in 95%ethanol,and determined using high-performance liquid chromatography(HPLC).He La,A549,SW620,HEK-293 T,THP-1,K562,Raw264.7 and MDA-MB-231 cell lines were used to investigate the morphology and mechanism of pyroptosis induced by L.erythrorhizon.The lactate dehydrogenase(LDH)release,propidium iodide(PI)/Hoechst double-staining,and pyroptosis reconstitution experiments were performed to study L.erythrorhizon-induced cell pyroptosis.Results:Compared with the death inhibitor,PI/Hoechst and LDH release experiments,we found that L.erythrorhizon induced pyroptosis.Recombination and western blot experiments confired that L.erythrorhizon induced GSDME cleavage,which drives pyroptosis.This phenomenon is conserved in several cancer cell lines that might be triggered by caspase family proteases.The mechanism of L.erythrorhizon inducing pyroptosis is widely found in tumor cells.Conclusion:Our findings not only explain how L.erythrorhizon triggers cancer cell pyroptosis,but also provide mechanistic insights to guide its clinical application in the future.

山东省部分市县麦田杂草麦家公Lithospermum arvense对苯磺隆的抗药性

采用温室盆栽法和培养皿法测定了山东省部分市县冬小麦田杂草麦家公Lithospermumarvense L.对苯磺隆的抗药性水平,以及其抗药性生物型乙酰乳酸合成酶(ALS)对苯磺隆的敏感性。温室盆栽结果显示,供试杂草对苯磺隆产生了不同程度的抗药性,其中胶州麦家公生物型抗性水平最高,抗性倍数为12.8倍;培养皿法测定结果也显示胶州麦家公生物型抗性水平最高,但抗性倍数为3.89倍。交互抗性测定结果表明,胶州抗性麦家公生物型对其他ALS抑制剂噻吩磺隆和苄嘧磺隆已产生不同程度的交互抗性,其中对噻吩磺隆的抗性倍数达到3.11倍。离体条件下,与敏感生物型ALS活力的抑制中浓度(IC50)相比较,胶州抗性麦家公生物型的IC50值是敏感麦家公的2.65倍。表明ALS敏感性降低可能是山东部分市县麦家公对苯磺隆产生抗药性的重要原因之一。

Screening and Identifying of Nephrotoxic Compounds in Lithospermum erythrorhizon Using Live-cell Fluorescence Imaging and Liquid Chromatography Coupled with Tandem Mass Spectrometry

In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon,it was separated into serial fractions according to their polarities.An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis.As a result,the primary fraction A05 and its secondary fractions C06 ＂C09 and C12 ＂C14 were found to have significant toxicity to LLC-PK1 cell line,as determined by the survive rate less than 20% after they were treated with these fractions.These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry.The main compounds in these fractions were tentatively identified to be acetylshikonin,isobutyrylshikonin,β,β＇-dimethyla-cryloylshikonin,and isovalerylshikonin,which may bring nephrotoxicity.

Orthogonal Experiment Design to Optimize the Pet Dog Dyeing Condition of Musa basjoo Siebold(MBS) & Lithospermum erythrorhizon(LE) Mixed Hair Dye

[Objectives] To optimize the pet dog dyeing process of plant hair dye. [Methods]Three factors affecting dyeing were selected including pH,concentration of dye and dyeing time. The L_9(3~3) orthogonal design was used to evaluate the hair coloring effect and the absorbance of cleaning solution after hair was dyed. [Results] The optimum pet dog dyeing process of plant hair dye was dye concentration of50 mg/mL,dye pH of 7. 5 and dyeing time of 1 h. [Conclusions] The coloring and fixing effects of the dye and dyeing process are good,which can lay a foundation for the development and utilization of plant dyes.

Clinical Effects of Lithospermum Ruderale Dosage and Using-time on Medicinal Abortion Induced by Mifepristone and Misoprostol

The clinical effects of dosage during the period of treatment of Lithospermum Ruderale extract-a kind of Chinese traditional herbal medicine-on medicinal abortion induced by mifepristone and misoprostol were studied. Lithospermum Ruderale extract was administrated 3 d before, 3 d after or 3 d before plus 3 d after the administration of misoprostol respectively. The dose of Lithospermum Ruderale extract was 50 g, 75 g or 100 g respectively. Thus 1 350 women of early pregnancy were grouped into 9 groups and observed. The results showed that the effects of Lithospermum Ruderale used 3 d before, and 3 d before plus 3 d after (6 days misoprostol were significantly better than those only used 3 d after misoprostol both for complete abortion and bleeding (P<0.05). The dosage between 50 g and 100 g made no significant difference in clinical effects. Therefore it is reasonable to use 50 g Lithospermum Ruderale before misoprostol to improve medicinal abortion.

Clinical Effects of Lithospermum Ruderal on Medical Abortion by Mifepristone

Early pregnancies(<42d)in 600 women were terminated respectively by mifepristone and Lithospermum Ruderal with misoprostol or mifepristone with misoprostol. The results showed that the rate of complete abortion(99.00%)of 300 women by mifepristone and Lithospermum Ruderal with misoprostol was significantly higher than that (92. 33%)of the other 300 women by mifepristone with misoprostol(P<0.01).The average length of time of embryo removal(54.01±7.55 h)and the rate of embryo removal (82.33%)within 6 hours after administration of misoprostol of the former were significantly shorter and higher than those(56.41±9.93 h and 72.67%) of the latter(P<0.05 respectively).The average length of the bleeding time of the former(7.01±3.11d)was significantly shorter than that(8.09±5.63d)of the latter(P<0.05).The recovery of the menstruation of both was not significantly different from each other(P>0.05).In all, the medical abortion by mifepristone and Lithospermum Ruderal with misoprostol was even more successful,with less bleeding time.Therefore it is essential to make researches on the auxiliary effects of Lithospermum Ruderal on the medical abortion by mifepristone with misoprostol.

Investigation on Adsorption of Lithospermum erythrorhizon onto Fungal Cell Wall Polysaccharides

A culture of Lithosperrnum erythrorhizon adsorbed on fungal cell wall polysaccharides, a novel bioadsorbent made from fungal cell wall, has been established in this paper. Three steps were involved in this immobilization. The first step was preparation of suspended plant cells from tightly aggregated plant cell clumps. The disassembled ratio of 0.715g·g^-1 (the disassembled cells over total cells) was obtained under optimum condition for the enzymatic reaction. Then, the adsorption of plant cells onto fungal cell wall polysaccharides was conducted and the saturated capacity of 12g cell per gram of carrier was obtained in adsorption immobilization. Finally, the culture of cells adsorbed on fungal cell wall polysaccharides was compared with that of cells entrapped in alginate or suspension cell culture. While exposed to in situ liquid paraffin extraction coupled with cell culture, the shikonin productivity of immobilized cells by adsorption was 10.67g·L^-1, which was 1.8 times of that in suspension culture and 1.5 times of that entrapped in alginate.

紫草植物染料对玫瑰永生花的染色工艺研究

为解决永生花化学染色污染、不安全的问题,以脱水脱色的玫瑰永生花为试材,利用天然紫草植物染料对其进行染色的单因素实验和正交实验,通过紫外—可见光谱法对紫草植物染料定性测试其最大吸收波长,并对花瓣的上染率以及染色后的花瓣的颜色特征值、K/S值等进行定量测试并分析,以花瓣上染率和染色深度K/S值为指标,研究天然紫草染料染色玫瑰永生花的最佳工艺条件,解析不同染色条件对染色的玫瑰永生花的颜色特性的影响,以期为其他植物染料对永生花的染色研究提供参考依据。结果显示:天然紫草植物染料的最大吸收波长为522 nm。天然紫草植物染料染色玫瑰永生花的最佳工艺为:染色时间为7 h,染料浓度为0.2%,温度为40℃,pH=3。此外,不同染色工艺下,玫瑰永生花的颜色特征值不同,呈现出不同的色彩。综上所述,天然紫草植物染料可应用于玫瑰永生花的染色中,整个工艺流程绿色环保、操作便捷,染色后的花朵色彩鲜艳、花型较好、整体效果逼真,可长久保存。

紫草外用治疗皮肤病的研究进展

紫草为传统常用中药,紫草有凉血活血、清热解毒、滑肠通便的作用。临床上常外用于皮肤病,治疗压疮疮疡、新生儿红臀、皮肤溃疡、皮肤瘙痒、湿疹皮炎、水火烫伤等病证。多以紫草油或紫草膏剂型使用,剂型单一。此文以紫草为主题词检索CNKI和维普等数据库,选取其中代表性的30篇文献,并对其进行分类归纳,总结了紫草治疗皮肤疾病的相关研究报道和临床应用,以期为临床更好地应用紫草,及以紫草为原料研究开发新药提供一定的理论参考。

紫草栓剂的制备

以热熔法制备紫草栓剂,以栓剂的颜色均匀度、硬度、色泽、完整度为评价指标,采用单因素实验筛选考察了不同基质组成比例、紫草加入量、过筛筛目、制备温度对紫草栓剂的影响。采用正交试验法优选紫草栓剂的最佳处方组成,确定制备紫草栓剂的最优条件。得到最佳条件:以聚乙二醇400、聚乙二醇4000和甘油作为栓剂基质,并按1∶2∶1的比例制备,加入7%经125μm(120目)筛网过筛的紫草粉,温度以70℃为宜。该制备方法设计合理,质量稳定,工艺可行。

肛周脓肿术后应用中药洗剂配合紫草油纱条引流临床观察

目的:观察中药洗剂配合紫草油纱条引流用于肛周脓肿术后的效果。方法:151例随机分为两组,对照组75例给予紫草油纱条引流,观察组76例给予益气养血洗液冲洗配合紫草油纱条引流。结果:观察组相较于对照组创面情况各指标均更低(P<0.05),两组术后7d相较于术后当日NPY、5-HT、P物质、IL-6、IL-8降低(P<0.05),同时间观察组比对照组更低(P<0.05)。结论:益气养血洗液冲洗配合紫草油纱条引流可促进肛周脓肿术后创面愈合,调节其血清致痛因子和炎症因子水平。

天然抗氧化剂的筛选

对７００余种中药和香辛料的抗氧化活性进行了大规模的筛选。发现有６４种植物有明显的抗氧化活性（保护系数４＞Ｐｆ≥２），２４种植物的抗氧化活性很强（保护系数Ｐｆ≥４）。其中有几种植物的抗氧化活性的筛选和研究，如荔枝草、降真香、紫草、血竭是以前未见报道过的。

天然染料紫草染色织物的迷彩伪装防护性能

采用传统的染色方法,用天然染料紫草对蚕丝和纯棉织物进行染色实验,研究了染色后这些织物的迷彩伪装性能。实验发现,用紫草染色的天然纤维织物不仅有很好的紫外防护性能,而且对可见近红外迷彩伪装也有很好的防护,其防护指标能达到军队的防护标准。紫草迷彩伪装性能可能是它的主要成分α萘醌的共轭电子跃迁的结果。

紫草抗氧化成分的提取及其活性测定

分别用石油醚、三氯甲烷、乙醇依次提取紫草成分，浓缩干燥后，用烘箱法及ＯＳＩ仪测定了３种提取物对油脂氧化稳定性的影响，发现石油醚提取成分（ＬＥ－ｐｅｔ）和三氯甲烷提取成分（ＬＥ－ｃｈｌ）有明显的抗氧化活性。在猪油中，ＬＥ－ｐｅｔ比ＶＥ的抗氧化活性强。

RP-HPLC法同时测定紫草中紫草素、异丁酰紫草素和β，β-二甲基丙烯酰紫草素的含量

目的:建立同时测定紫草中紫草素、异丁酰紫草素和β,β-二甲基丙烯酰紫草素含量的方法。方法:采用 RP-HPLC法,Zorbax Eclipse XDB-C_(18)柱(250 mm×4.6 mm,5μm);流动相为含0.5%冰醋酸及0.3%三乙胺的乙腈溶液(A)和含0.5%冰醋酸及0.3%三乙胺的水溶液(B),梯度洗脱,洗脱程序为:0 min(67%A),5 min(70%A),15 min(75%A),30 min(75%A);流速1.0 mL·min^(-1);检测波长516 nm,柱温30 ℃。结果:紫草素、异丁酰紫草素和β,β-二甲基丙烯酰紫草素浓度分别在1.2～12.2μg·mL^(-1)(r=0.9999)、2.5～24.8μg·mL^(-1)(r=0.9998)、11.3～112.8μg·mL^(-1)(r=0.9998)范围内与峰面积呈良好线性关系(n=5)。方法回收率(n=9)分别为100.3%,98.6%,98.4%。结论:该方法简便、快速、准确,重复性好,可用于紫草中紫草素、异丁酰紫草素和β,β-二甲基丙烯酰紫草素含量的同时测定。

梓木草的化学成分

AIM: To study the chemical constituents of Lithospermum zollingeri.METHOD: Many kinds of column chromatography were used to isolate the compounds from the EtOH ext.of Lithospermum zollingeri DC.The chemical constituents of the plant were identified by means of MS,(（）1H NMR),(（）13C NMR),respectively,in some cases by direct comparison with authentic samples.RESULT:Seven compounds were isolated from the EtOH ext.of Lithospermum zollingeri.Their structures were identified as acacetin-7-O-β-D-glucopyranoside（Ⅰ）,hesperidin（Ⅱ）,apigenin-7-O-β-D-glu-copyranoside（Ⅲ）,luteolin-7-O-β-D-glucopyranoside（Ⅳ）,chrysoeriol-7-O-β-D-glucopyranoside（Ⅴ）,daucosterol（Ⅵ） and β-sitosterol（Ⅶ） respectively,based on their spectroscopic data.CONCLUSION:All the compounds were isolated from this plant for the first time.

紫草提取物抗氧化作用研究

考察了紫草提取物在体外和体内的抗氧化活性。以合成天然抗氧化剂二叔丁基对甲酚(BHT)和天然抗氧化剂α-生育酚(α-tocopherol)为对照,用OSI方法测定了紫草石油醚提取物和氯仿提取物在猪油中的抗氧化作用,并首次考察了二者对小鼠血清中超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平的影响。结果表明,添加质量分数为0.1%的紫草石油醚和氯仿提取物,可分别使猪油的氧化诱导期由4.5 h延长至52.4 h和52.2 h。小鼠灌服紫草石油醚和氯仿提取物21 d后,其血清中SOD活性均显著高于正常组(P<0.05或P<0.01),而MDA含量显著低于正常组(P<0.05或P<0.01),表现出体内抗氧化活性,证明紫草石油醚和氯仿提取物具有较强抗氧化活性,可显著抑制猪油氧化,并可明显提高小鼠体内SOD的活性,降低MDA含量。该研究内容的新颖性,已为教育部科技查新工作站(G05)2008年2月28日出具的的第DHU2008-019号《科技查新报告》所证实。

紫草提取物的体外抗氧化活性研究

为研究紫草色素(LEP)的体外抗氧化活性,采用常规回流方法提取LEP,测定了LEP对超氧自由基(O2)和1,1-二苯基-2-苦苯肼自由基(DPPH.)的清除能力,及其对β-胡萝卜素/亚油酸自氧化体系的抑制作用。结果表明,LEP对DPPH.和O2均有较强的清除能力,并且对β-胡萝卜素/亚油酸自氧化体系有明显的抑制作用,说明紫草的药理作用可能与LEP较强的抗氧化能力有关。

紫草的药理作用与应用研究进展

本文对近年来紫草在抗菌、抗病毒、抗生育及避孕、抗炎及镇痛、抗肿瘤及免疫调节、止血。

从细胞自噬角度探讨紫草多糖抑制H22肝癌实体瘤细胞增殖的作用机制

目的:探讨紫草多糖是否通过诱导自噬而抑制H_(22)肝癌实体瘤细胞的增殖。方法:将40只KM小鼠随机分为模型组、环磷酰胺组(阳性药物,30 mg/kg,ip,每4 d给药1次)和紫草多糖低、高剂量组(100、300 mg/kg,ig,每天给药1次),每组10只。各组小鼠均腋下接种H_(22)细胞株复制H_(22)肝癌实体瘤模型,造模同时各给药组小鼠给予相应药物,模型组小鼠ig等体积生理盐水。末次给药后,记录并测定小鼠体质量、瘤质量系数和胸腺、脾脏指数;采用实时荧光定量聚合酶链式反应法测定小鼠瘤组织中自噬相关基因Atg5、Beclin1 m RNA表达水平;Western blot法测定瘤组织中自噬微管相关蛋白1轻链3A/B(LC3A/B)蛋白表达水平。结果:紫草多糖可显著降低H_(22)实体瘤小鼠的瘤质量系数,抑瘤率达34.7%;可显著上调瘤组织中自噬相关基因Atg5、Beclin1 m RNA的表达和LC3A/B蛋白的表达,较模型组差异均有统计学意义(P<0.05或P<0.01)。结论:紫草多糖通过促进肝癌细胞的自噬,从而抑制H_(22)肝癌实体瘤细胞的增殖、延缓肿瘤生长。