Lignans from Patrinia scabiosaefolia improve insulin resistance by activating PI-3K/AKT pathway and promoting GLUT4 expression

Patrinia scabiosaefolia,is used as wild vegetable in China for more than 2000 years,with a variety of pharmacological activities,including anti-inflammatory,anti-tumor and hypoglycemic.Based on our ongoing research on chemical constituents and hypoglycemic activity of P.scabiosaefolia,4 lignan compounds,(+)-isolariciresinol(1),7R,7’R,8S,8’S-(+)-neo-olivil-4-O-β-D-glucopyranoside(2),4-O-methylcedrusin(3)and patrinian A(4),were isolated and identifi ed.The hypoglycemic activity showed that compounds 2 and 3 could extremely signifi cantly improve insulin resistance at 100(P<0.001),50(P<0.001)and 25μmol/L(P<0.01)in IR 3T3-L1 cells.While compound 4 only promoted glucose uptake by IR 3T3-L1 cells at 100μmol/L(P<0.01).Western blotting experiments showed that compounds 2 and 4 up-regulated the protein expressions of p-IRS,PI-3K,p-AKT and glucose transporter 4(GLUT4),and promoted the transcription of GLUT4 mRNA.Therefore,the mechanisms of compounds 2 and 4 were presumed to improve IR by activating PI-3K/AKT signaling pathway.

Two New Antibacterial Iridoids from Patrinia rupestris

Two new iridoids （1 and 2） were isolated from the roots of Patrinia rupestris （Pail.） Juss. Their structures were elucidated by spectroscopic methods. Compounds 1 and 2 exhibited strong antibacterial activities against Eschecichia coli and Staphylococcus aureus, respectively.

A New Alkaloid from Patrinia scabra

A new alkaloid has been isolated from the root of Patrinia scabra. Its structure was elucidated as 2-acetamido-3-phenyl propyl 2-benzamido-3-phenyl propionate by extensive spectr- oscopic analysis.

Two New Dihydroflavanoids from Patrinia villosa Juss

Two new dihydroflavonoids were obtained from the traditional Chinese medicinal herb Patrinia villosa Juss. Their structures were elucidated as （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6, 8-di （γ, γ-dimethylallyl） flavanone （1） and （2S）-5, 7, 2＇, 6＇-tetrahydroxy-6-1avandulylated flavanone （2） by spectroscopic methods including UV, IR, HR-TOF-MS, 1D NMR and 2D NMR techniques.

Scabrous patrinia root inhibits circulating tumor cells in differentiated thyroid carcinoma–a clinical observation

Objective To observe the clinical inhibition of circulating tumor cells(CTCs) in differentiated thyroid carcinoma(DTC) by the extract of scabrous patrinia root(Huikangling).Methods Eighty-seven DTC patients tested positive for CTCs were randomly divided into two groups; all patients were treated with oral levothyroxine sodium in accordance with the DTC endocrine inhibition treatment criteria. Patients(n = 45) in the treatment group were provided the standard endocrine therapy along with oral Huikangling(0.4 g/tablet, 0.4 g × 3/time, 3 times/day, 12 weeks). Patients(n = 42) in the control group were only provided the standard therapy. After 4 and 12 weeks, CTCs in the blood were detected by flow cytometry.Results After 4 weeks of oral Huikangling treatment, CTCs were detected in 18(40%) and 29(69%) patients in the treatment and control groups, respectively; the difference was statistically significant(χ2 = 8.49, P < 0.05). After 12 weeks, CTCs were detected in 7(15.6%) and 17(44.7%) patients in the treatment and control groups, respectively; the difference was statistically significant(χ2 = 5.68, P < 0.05). Follow-up evaluation revealed two patients with lung metastasis and one patient with bone metastasis in the control group; one patient showed lateral neck lymph node metastasis without local recurrence in the treatment group.Conclusion Huikangling treatment reduces the number of CTC-positive DTC cases; however, further studies are needed to elucidate the underlying mechanisms.

Two New Dihydroflavanoids from Patrinia vittosa Juss (II)

Two new dihydroflavonoids named villosin A and viUosin B were obtained from the traditional Chinese medicinal herb Patrinia viUosa Juss. Their structures were elucidated as （2S）-5, 2＇, 6＇-trihydroxy-2＂, 2＂-dimethylpyrano [5＂, 6＂： 6, 7]flavanone 1 and （2S, 3＂S）-5, 2＇, 6＇- trihydroxy-3＂-γ, γ-dimethylallyl-2＂, 2＂-dimethyl-3＂, 4＂-dihydropyrano [5＂, 6＂： 6, 7] flavanone 2 by spectroscopic methods including UV, IR, HR-TOF- MS, 1D NMR and 2D NMR techniques.

Network pharmacology-based screening of the active ingredients and potential targets of the Patrinia villosa for application to Alzheimer’s disease

Alzheimer’s disease(AD)is a progressive neurodegenerative disorder associated with global deterioration of intellectual function,including memory,learning,orientation,language,comprehension and judgment.Given the prevalence of AD and the lack of effective long-term therapies,there is a pressing need to discover viable leads that can be developed into clinically approved drugs with disease-modifying effects.The analysis of current reported literatures confirms the importance of Patrinia villosa J.as candidate against AD.Hence,the compound-disease target network based on the methods of network pharmacology prediction was established in this paper.The aim is to explore potential compounds with marked effect on AD in P.villosa.The result showed that the most effective compound in this plant might be threo-(7R,8R)-guaiacyl-glycerol-β-O-4′-dihydroconiferyl ether,and the most potential target might be Maltose-binding periplasmic.

Inhibitory effect of Patrinia scabiosaefolia on acute pancreatitis

瞄准：在缩胆囊素(CCK ) 上调查 Patrinia scabiosaefolia (PS ) 的效果在老鼠的导致 octapeptide 的尖锐胰腺炎(AP ) 。方法：称 240-260 g 的 Wistar 老鼠被划分成三个组：(1 ) 正常的盐对待的组；(2 ) 在哪个 PS 口头上地被管理，有在 100 mg/kg 组的 PS 的处理在 1， 3 和 5 h，和这个全部以后由 75 microg/kg CCK octapeptide 的下的管理列在后面三次过程为 5 d 被重复；(3 ) 与盐的组，协议与在处理一样在是一起的处理与 PS 组织。我们决定了胰腺的重量 / 身体重量比率，胰腺的 HSP60 的层次， HSP72 和支持 inflammatory cytokines 的分泌物。重复 CCK octapeptide 处理导致了试验性地导致的胰腺炎的典型实验室调查结果。结果：PS 在 CCK 导致 octapeptide 的 AP 减少了胰腺的重量 / 身体重量比率，浆液淀粉酶和脂肪分解酵素的层次，和支持 inflammatory cytokines 的禁止的表情。而且， PS 预告的处理增加了 HSP60 和 HSP72 的胰腺的层次。结论：有 PS 的预告的处理在 CCK 导致 octapeptide 的 AP 上有反煽动性的效果。

败酱草水提物通过调控circ_0000936/miR-665抑制胶质瘤细胞增殖、迁移及侵袭

目的:探讨败酱草水提物对胶质瘤细胞生物行为的影响及其可能作用机制。方法:收集2019年2月至2020年1月期间于本院接受手术切除并经病理证实的31例胶质瘤组织,qRT-PCR法检测正常脑组织与胶质瘤组织中circ_0000936、miR-665的表达量;Pearson法分析胶质瘤组织中circ_0000936和miR-665表达量的相关性;体外培养人胶质瘤细胞LN229,不同浓度的败酱草(0.5 mg/mL、1.0 mg/mL、2.0 mg/mL)处理LN229细胞,si-NC、si-circ_0000936分别转染至LN229细胞,pcDNA-circ_0000936转染至LN229细胞后加入2.0 mg/mL败酱草处理细胞;CCK-8、平板克隆形成实验、Transwell小室实验与流式细胞术分别检测细胞增殖、克隆形成、迁移、侵袭及凋亡能力;双荧光素酶报告基因实验检测circ_0000936与miR-665的靶向关系;Western blot法检测Bax、Bcl-2蛋白表达量。结果:胶质瘤组织中circ_0000936的表达量高于正常脑组织(P<0.05),而miR-665的表达量低于正常脑组织(P<0.05);circ_0000936和miR-665呈负相关(r=-0.980,P<0.001);败酱草能够以浓度依赖性方式降低细胞活力、细胞克隆形成率、Bcl-2蛋白水平和circ_0000936的表达量(P<0.05),迁移及侵袭细胞数减少(P<0.05),而凋亡率、Bax蛋白水平和miR-665的表达量升高(P<0.05);转染si-circ_0000936后miR-665的表达量升高(P<0.05),细胞活力、细胞克隆形成率和Bcl-2蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),凋亡率和Bax蛋白水平升高(P<0.05);转染pcDNA-circ_0000936能够逆转败酱草对LN229细胞生物学行为的作用。结论:败酱草水提物可通过调控circ_0000936/miR-665诱导胶质瘤细胞凋亡,并阻碍增殖、迁移及侵袭。

Patrinia scabiosaefolia Inhibits Growth of 5-FU-Resistant Colorectal Carcinoma Cells via Induction of Apoptosis and Suppression of AKT Pathway

Objective: To investigate the effects of ethanol extract of Patrinia scabiosaefolia(EEPS) on chemo-resistance of colorectal cancer cells(CRC) and explore the possible molecular mechanisms. Methods: 5-fluorouracil(5-FU)-resistant human colorectal carcinoma cell line(HCT-8/5-FU) and its parental cells HCT-8 were treated with EEPS(0, 0.25, 0.50, 1 or 2 mg/mL), or 5-FU(0, 100, 200, 400, 800 or 1600 μmol/L). The 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assay was performed to evaluate the cell viability. Cell density was observed by phase-contrast microscope, cell counting and colony formation assay were used to determine the cell proliferation of HCT-8/5-FU cells treated with 0, 0.5, 1 or 2 mg/mL EEPS. Cell apoptosis was determined by Hoechst staining. Western-blot was performed to detect the phosphorylation of AKT as well as the protein expression level of B-cell CLL/lymphoma 2(Bcl-2) and Bcl-2-associated X protein(Bax). Results: Compared with HCT-8 cells, MTT assay results indicated that HCT-8/5-FU cells were resistant to 5-FU treatment(P<0.05), and sensitive to EEPS treatment(P>0.05). Moreover, compared with untreated HCT-8/5-FU cells, 1 and 2 mg/mL of EEPS treatment significantly reduced cell density, cell number, inhibited cell survival(P<0.05), and induced apoptosis in HCT-8/5-FU cells. Furthermore, 1 and 2 mg/mL of EEPS significantly decreased the phosphorylation level of p-AKT and Bcl-2 protein expression, and increased the expression of Bax protein(P<0.05). Conclusion: EEPS is a promising therapeutic agent that may overcome chemo-resistance in cancer cells, likely through suppression of the AKT pathway and promotion of cancer cell apoptosis.

白花败酱草化学成分与药理活性研究进展

白花败酱草是败酱属植物,分布广泛,植物资源量大。在我国,白花败酱草具有悠久的药用历史,作为一味传统中药在多种方剂中出现。在中医临床应用中,白花败酱草在治疗呼吸系统、消化系统、泌尿生殖系统、妇科、皮肤科等多学科疾病的治疗实践中得到了广泛应用。目前,对于白花败酱草的化学成分研究进展颇丰,已提取分离出81种单体成分,包括苯丙酸类、黄酮类、萜类等多种结构类型。同时,药理学研究发现白花败酱草具有多种明确的药理活性,包括抗肿瘤、消炎镇痛、抑菌以及保护肝脏等,并对其作用机制进行了初步研究探讨。该文在临床功效、化学成分、药理作用等方面对白花败酱草进行了综述,归纳总结现有已发表的研究成果,为其临床应用和进一步研究提供依据和基础。

Chemical Composition,Anticancer,Anti-neuroinflammatory,and Antioxidant Activities of the Essential Oil of Patrinia scabiosaefolia

Objective：To study the chemical composition,anticancer,anti-neuroinflammatory,and antioxidant activities of the essential oil of Patrinia scabiosaefolia（EO-PS）.Methods：Patrinia scabiosaefolia was analyzed by gas chromatography-mass spectrometry.Eight human carcinoma cell lines,including SGC-7901,AGS,Hep G2,HT-29,HCT-8,5-FU/HCT-8,He La,and MDA-MB-231,were assessed by methylthiazolyldiphenyltetrazolium bromide（MTT） assay.Anti-neuroinflammatory activity was assessed by production of interleukin（IL）-1β and IL-6 induced by lipopolysaccharide in BV-2 cells（microglia from mice）.The antioxidant activity was evaluated with a 2,2-diphenyl-1-picrylhydrazyl（DPPH） scavenging assay.Results：Forty-four components,representing 83.919% of the total oil,were identified in the EO-PS.The major constituents were caryophyllene oxide（12.802%）,caryophyllene（6.909%）,α-caryophyllene（2.927%）,β-damascenone（3.435%）,calarene（5.621%）,and phenol（3.044%）.The MTT assay showed that the EO-PS exhibited significant dose-dependent growth inhibition in the 50–200 μg/m L dilution range.The EO-PS exhibited a dose-dependent scavenging activity against the DPPH radical,with an half of maximal inhibitory concentration 1.455 mg/m L.Conclusion：The EO-PS possesses a wide range of antitumor,anti-neuroinflammatory and antioxidant activities,suggesting that it may be a good candidate for further investigations of new bioactive substances.

败酱草质量研究

目的:建立败酱草的薄层鉴别方法(TLC)及HPLC测定东莨菪内酯含量测定方法,为败酱草的质量研究提供依据。方法:采用TLC鉴别法,HPLC作为含量测定方法,色谱柱:Ultimate Plus C_(18)(4.6 mm×250 mm,5μm);流动相:乙腈-0.5%冰醋酸(15∶85);柱温:25℃;流速:1 mL·min^(-1);检测波长:343 nm。结果:薄层鉴别中检出与对照药材相同颜色的荧光斑点;东莨菪内酯在0.8384~20.96μg·mL^(-1)范围内线性关系良好(函数关系为Y=39.233X+7.0215,r=0.9999),回收率为91.0%~97.9%,均值为95.4%,RSD(n=6)为3%。结论:本方法鉴别专属性较好,含量测定方法专属性、稳定性、重现性、准确度良好,可用于败酱草的质量控制。

Triterpenoid saponins from Patrinia scabra

AIM: To study the chemical constituents and bioactivity of the roots of Patrinia scabra Bunge. METHODS: The chemical constituents were isolated using various chromatographic methods, and the structures were elucidated on the basis of spectral analysis and chemical methods. In addition, cytotoxic activities toward HepG2 cells were tested by the MTT method. RESULTS: A new triterpenoid saponin, 3-O-(4'-isovaleryl)-O-β-D-xylose-12,30-dihydroxy-oleanane-28,13-lactone-22-O- β-D-glucoside(1), along with two known triterpenoid saponins, acanthopanax saponin CP3(2) and foetoside C(3), were isolated. CONCLUSION: The aglycone of compound 1 was a new skeleton derivative of oleanolic acid. Compound 2 showed strong cytotoxicity to HePG2 cells(IC50 1.49 μmol?L-1).

不同环境因素对苦菜提取液抑菌稳定性的影响

随着时代的发展,人们对食品安全的关注度上升,因此对天然防腐剂的开发和应用成为近几年的研究热点。文章在前期研究的基础上,继续对苦菜提取液抑菌稳定性进行研究。以大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、绿脓杆菌为供试菌种,探讨温度、pH、紫外线、金属离子、蔗糖、保存时间等对苦菜提取液抑菌活性的影响。结果表明,苦菜提取液对热处理、紫外线、保存时间均具有较好的稳定性;pH为7.0时,苦菜提取液的抑菌效果最好;Ca^(2+)、Fe^(2+)、Fe^(3+)有增强苦菜提取液抑菌能力的作用;随着蔗糖浓度的增大,提取液的抑菌活性有所增强。苦菜提取液的抑菌活性较稳定,可适应多种加工条件,作为天然的抑菌防腐剂使用,有较好的应用前景。

黄花败酱茎秆二氯甲烷萃取相对人白血病细胞K562增殖及分化的影响

目的:探讨黄花败酱茎秆乙醇提取物的二氯甲烷萃取相(DPSS)对K562细胞增殖抑制和诱导分化作用及其相关作用机制。方法:不同浓度DPSS(0、25、50、100和200μg/ml)分别作用于细胞24、48和72 h后,采用MTT法检测K562细胞增殖情况;应用流式细胞术检测不同浓度DPSS对K562细胞凋亡、周期阻滞的影响;应用瑞氏-吉姆萨染色观察DPSS作用后K562细胞形态学的变化并应用流式细胞术验证分化相关抗原CD33、CD11b的表达。结果:与对照组比较,加药组各浓度DPSS均对K562细胞的增殖有明显抑制作用,并且存在时间剂量依赖性(r=-0.96);细胞周期分析结果表明,随着DPSS浓度升高,G2/M期细胞增多(r=0.88),细胞被阻滞在G_(2)/M期;流式细胞术结果显示,当200μg/ml DPSS作用48 hK562细胞凋亡率最高;瑞氏-吉姆萨染色结果显示,DPSS作用后K562细胞出现胞体增大,包浆量增多,核浆比减小,核染色质粗糙等分化表现;细胞分化抗原检测发现,DPSS作用后CD33、CD11b呈阳性表达。结论:DPSS可以通过周期阻滞诱导K562细胞凋亡、抑制其增殖,并且能够诱导K562细胞向单核细胞分化,具有潜在抗白血病细胞的作用。

响应面法优选败酱草多糖的脱色工艺及其体外抗氧化活性研究

在单因素试验基础上,采用四因素三水平的Box-Behnken试验设计,建立各因素与脱色率、蛋白脱除率和多糖保留率之间的数学模型,得出败酱草多糖的最佳脱色工艺;对比大孔树脂脱色前后败酱草多糖的理化特征以及对ABTS、DPPH、超氧自由基和羟自由基等的体外清除活性。结果表明:大孔树脂HPD-450对败酱草多糖脱色效果最佳,其最优工艺为:振荡时间1 h,树脂用量6.6 g·10 mL^(-1)败酱草多糖溶液,pH 3.2,温度35.0℃。在此条件下,脱色率、蛋白脱除率和多糖保留率分别为85.16%、90.30%和59.60%。与脱色前相比,HPD-450脱色后多糖中优势糖为葡萄糖和半乳糖,总糖(纯度)和还原糖含量得到了提高,部分蛋白质也被去除。体外抗氧化活性结果显示,脱色后败酱草多糖对羟自由基的清除能力明显增强,当浓度为4.00 mg·mL^(-1)时,其对羟自由基的清除率达98.90%,接近于Vc。脱色提高了败酱草多糖的纯度,其中所含葡萄糖、半乳糖的含量明显增加,脱色后败酱草多糖对羟自由基的清除率显著增强。

基于败酱草多糖的能量冲剂的开发

为研发败酱草多糖的能量冲剂,通过单因素和正交试验分别确定加入多糖、可溶性淀粉、甜蜜素的量,从而优选出冲剂的最佳配方。结果表明,败酱草多糖、可溶性淀粉、甜蜜素的最佳加入量分别为1.30、0.50、0.02 g,使用75%乙醇作为润湿剂可制得成型性和流动性较好且不易黏连的败酱草多糖颗粒剂。

败酱草治疗慢性盆腔炎的机制研究

目的:应用网络药理学方法流程筛选败酱草的主要活性成分,预测该主要活性成分治疗慢性盆腔炎(CPID)的潜在作用靶点及信号通路,探讨其作用机制,并用分子对接预计分子间结合能,探讨结合的优良性。方法:在TCMSP、SwissTarget两个数据库上筛选获得败酱草的潜在活性成分和活性成分对应的靶点;慢性盆腔炎的疾病靶点可通过在线数据库OMIM、Drugbank两个数据库获得,应用Uniprot数据库对成分靶点名称规范化。通过STRING在线数据库获得各靶点相互作用关系;Cytoscape3.9.1软件对靶点关系进行可视化,构建中药-活性成分-靶点-疾病网络图和靶点蛋白-蛋白互作网络图(PPI)。利用Cytoscape软件中的插件筛选核心成分及疾病靶点。利用在线数据库DAVID对靶点进行KEGG通路富集分析。结果:败酱草治疗慢性盆腔炎的核心活性成分包括山奈酚、槲皮素等;PPI互作网络显示关键蛋白涉及PTGS2、IL10、TNF等;KEGG分析结果显示,这些靶点主要富集映射在TNF等信号通路上。结论:该研究预测了败酱草治疗慢性盆腔炎的主要作用机制,说明败酱草可能通过“多成分、多靶点、多通路”发挥抗慢性盆腔炎作用。

基于网络药理学和分子对接技术分析大血藤-败酱草抗结直肠癌的作用机制

目的采用网络药理学和分子对接技术分析大血藤-败酱草抗结直肠癌的分子机制。方法首先采用公共数据库挖掘大血藤-败酱草的相关活性成分和靶点以及结直肠癌的相关靶点,筛选出结直肠癌治疗的潜在靶点。然后进行中药-成分-靶点网络和蛋白质-蛋白质相互作用(PPI)网络分析,采用基因本位(GO)和京都基因与基因组百科全书(KEGG)通路富集分析治疗结直肠癌涉及的分子功能、细胞成分、生物过程及相关通路,并构建靶点-通路网络,筛选结直肠癌治疗的关键靶点。最后采用分子对接技术评价关键成分与靶点的亲和力。结果大血藤-败酱草治疗结直肠癌的成分主要是芹菜素、槲皮素、大黄素等,涉及促分裂原活化的蛋白激酶(MAPK)1、MAPK3、蛋白激酶B丝氨酸/苏氨酸激酶1(AKT1)、磷脂酰肌醇-4,5二磷酸3-激酶催化亚单位α(PIK3CA)等多个靶点。GO分析主要富集于蛋白质磷酸化、蛋白激酶活性、转录调控复合物等方面;KEGG通路富集分析显示,癌症通路、表皮生长因子受体(EGFR)-酪氨酸激酶抑制剂(TKI)耐药性、磷脂酰肌醇-3-羟激酶(PI3K)/蛋白激酶B(AKT)信号通路等均是结直肠癌治疗的重要通路。芹菜素、槲皮素、大黄素与MAPK1、MAPK3和PIK3CA均有强烈亲和力,与AKT1亲和力较好,芹菜素-PIK3CA、槲皮素-MAPK1、大黄素-PIK3CA能够形成相对稳定的复合物。结论大血藤-败酱草可能通过抑制MAPK1、MAPK3、和PIK3CA靶蛋白的表达,通过调控PI3K/AKT、MAPK信号通路介导的细胞增殖、迁移和凋亡等过程发挥抗结直肠癌作用。