Targeting colorectal cancer with Herba Patriniae and Coix seed:Network pharmacology,molecular docking,and in vitro validation

BACKGROUND Herba Patriniae and Coix seed(HC)constitute a widely utilized drug combination in the clinical management of colorectal cancer(CRC)that is known for its diuretic,anti-inflammatory,and swelling-reducing properties.Although its efficacy has been demonstrated in a clinical setting,the active compounds and their mechanisms of action in CRC treatment remain to be fully elucidated.AIM To identify the active,CRC-targeting components of HC and to elucidate the mechanisms of action involved.METHODS Active HC components were identified and screened using databases.Targets for each component were predicted.CRC-related targets were obtained from human gene databases.Interaction targets between HC and CRC were identified.A“drug-ingredient-target”network was created to identify the core components and targets involved.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were conducted to elucidate the key pathways involved.Molecular docking between core targets and key components was executed.In vitro experiments validated core monomers.RESULTS Nineteen active components of HC were identified,with acacetin as the primary active compound.The predictive analysis identified 454 targets of the active compounds in HC.Intersection mapping with 2685 CRC-related targets yielded 171 intervention targets,including 30 core targets.GO and KEGG analyses indicated that HC may influence the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway.Molecular docking showed that acacetin exhibited an optimal interaction with AKT1,identifying PI3K,AKT,and P53 as key genes likely targeted by HC during CRC treatment.Acacetin inhibited HT-29 cell proliferation and migration,as well as promoted apoptosis,in vitro.Western blotting analysis revealed increased p53 and cleaved caspase-3 expression and decreased levels of p-PI3K,p-Akt,and survivin,which likely contributed to CRC apoptosis.CONCLUSION Acacetin,the principal active compound in the HC pair,inhibited the proliferation and migration of HT-29 cells and promoted apoptosis through the PI3K/Akt/p53 signaling pathway.

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.

Inhibitory effect of Patrinia scabiosaefolia on acute pancreatitis

AIM： To investigate the effect of Patrinia scabiosaefolia （PS） on the cholecystokinin （CCK） octapeptide-induced acute pancreatitis （AP） in rats.METHODS： Wistar rats weighing 240-260 g were divided into three groups： （1） Normal saline-treated group; （2） treatment with PS at 100 mg/kg group, in which PS was administered orally, followed by subcutaneous administration of 75μg/kg CCK octapeptide three times after 1, 3 and 5 h, and this whole procedure was repeated for 5 d; （3） treatment with saline group, in which the protocols were the same as in treatment group with PS. We determined the pancreatic weight/ body weight ratio, the levels of pancreatic HSP60, HSP72 and the secretion of pro-inflammatory cytokines. Repeated CCK octapeptide treatment resulted in the typical laboratory findings of experimentally induced pancreatitis.RESULTS： PS reduced the pancreatic weight/body weight ratio, the levels of serum amylase and lipase, and inhibited expressions of pro-inflammatory cytokines in the CCK octapeptide-induced AP. Furthermore, PS pretreatment increased the pancreatic levels of HSP60 and HSP72.CONCLUSION： Pretreatment with PS has an antiiinflammatory effect on CCK octapeptide-induced AP.

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 spectroscopic 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.

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。

败酱草及其近缘种叶绿体全基因组分析和DNA条形码构建

目的:基于败酱类药材基原植物的叶绿体基因组序列分析开发特定的DNA条形码,准确鉴别败酱草及其近缘种。方法:采用Illumina高通量测序技术对败酱科败酱属植物白花败酱、黄花败酱和异叶败酱的叶绿体基因组进行测序;采用生物信息学软件对基因组进行组装注释、特征分析、序列比较和系统发育分析;基于叶绿体基因组高突变区构建特异性DNA条形码进行验证。结果:3种败酱属植物叶绿体基因组呈四分体结构,全长分别为159585、158919、158919 bp,编码的基因数分别为130、132、132,包含8个rRNA基因和37个tRNA基因,蛋白质编码基因数分别为85、87、87。ccsA、ndhG、rpl23、ndhF序列作为特异性DNA条形码成功扩增16份样品,黄花败酱、白花败酱和少蕊败酱聚类在同一支,异叶败酱和糙叶败酱聚类在另一支。通过ccsA、ndhG、ndhF序列可进一步鉴别糙叶败酱和异叶败酱。结论:ccsA、ndhG、rpl23、ndhF序列可作为补充特异性DNA条形码区分败酱草及其近缘种。

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。

糙叶败酱中獐牙菜苷的高速逆流色谱分离及其抗肿瘤活性研究

目的:采用高速逆流色谱法(HSCCC)对糙叶败酱中的环烯醚萜类化合物獐牙菜苷进行分离纯化,并对其体外抗肿瘤活性进行初步评价。方法:HSCCC的溶剂体系为正丁醇-乙酸乙酯-水(5∶2∶5),上相为固定相,下相为流动相,正进正转,分离温度25℃、转速850 r/min、流动相流速2.0 mL/min、进样浓度10 mg/mL、进样量15 mL、检测波长240 nm,并采用细胞计数试剂盒-8(CCK-8)法检测糙叶败酱对5种不同肿瘤细胞的增殖抑制作用。结果:从糙叶败酱中分离得到了纯度大于92%的獐牙菜苷。獐牙菜苷对人源肝癌细胞HepG2、结肠癌细胞SW480、肺癌细胞A549、人慢性髓原白血病细胞K562和宫颈癌Hela细胞这5种肿瘤细胞的增殖均有较强的抑制作用,且呈现明显的剂量依赖关系和时间依赖关系。结论:本研究为应用HSCCC高效分离纯化糙叶败酱中獐牙菜苷提供了技术支持,也为其抗肿瘤活性研究提供了思路。

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.

Triterpenoid saponins from Patrinia scabra

AIM： To study the chemical constituents and bioactivity of the roots ofPatrinia 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-28,13-1actone-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 cyto- toxicity to HePG2 cells （IC50 1.49 umol.L-1）.

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。

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。

败酱草提取液对仔兔脂多糖肠炎模型的修复作用

为研究败酱草提取液对脂多糖所致的仔兔肠炎的修复作用。选取40只仔兔,随机分为5组,空白组、模型组、败酱草提取液低、中、高剂量组。空白组腹腔注射2 mg/kg的生理盐水,其余各组仔兔通过腹腔注射2 mg/kg脂多糖(LPS)建立仔兔肠炎模型,连续3 d,每天注射1次。败酱草提取液低、中、高剂量组依次按1、3、5 mL/kg在饮水中添加败酱草提取液,其余组正常饮水,连续给药15 d,每天1次。通过苏木精-伊红(HE)染色从小肠的绒毛高度、隐窝深度和绒隐比(V/C)来研究败酱草提取液对脂多糖诱导仔兔肠炎的修复作用。结果表明,与空白组相比,模型组小肠的绒毛高度、V/C值均显著低于空白组且隐窝深度均显著高于空白组(P<0.05)。高剂量组小肠的绒毛高度和绒隐比均显著高于中剂量组(P<0.05)。研究表明,按5mL/kg在饮水中添加败酱草提取液可有效修复仔兔脂多糖模型。

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。