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.

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.

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.

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.

白花败酱草总黄酮对实验性急性肺损伤模型大鼠的肺保护作用及其机制

目的:探讨白花败酱草总黄酮(total flavoniods from Patrina villosa Juss,PJF)对实验性急性肺损伤(acute lung injury,ALI)模型大鼠的肺保护作用及其潜在机制。方法:用气管内滴注5 mg/kg脂多糖(lipopolysaccharide,LPS)构建ALI大鼠模型。将60只雄性SD大鼠随机分为对照组、LPS组、LPS+低剂量(100 mg/kg)PJF组和LPS+高剂量(300 mg/kg)PJF组(后2组在ALI造模前1 h给予PJF灌胃),每组15只。造模后24 h,收集支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)和肺组织。HE染色显示肺组织形态;干湿称重法测量肺组织的湿/干重比;伊文思蓝染色评估肺组织中上皮屏障的通透性;ELISA检测BALF中炎症因子肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和IL-6含量,以及肺组织中氧化应激指标超氧化物歧化酶(superoxide dismutase,SOD)、髓过氧化物酶(myeloperoxidase,MPO)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性及丙二醛(malondialdehyde,MDA)含量;免疫印迹法检测肺组织中C/EBP同源蛋白(C/EBP homologous protein,CHOP)、葡萄糖调节蛋白78(glucose-regulated protein 78,GRP78)和X框结合蛋白1(X-box binding protein 1,XBP1)蛋白表达。结果:与对照组比较,LPS组大鼠肺组织呈现肺泡结构不清和大量炎症细胞浸润,ALI评分和肺组织的湿/干重比显著升高(P<0.05),BALF中IL-6、IL-1β和TNF-α水平,以及肺组织中MDA含量、MPO活性及CHOP、GRP78和XBP1蛋白表达水平显著升高(P<0.05),肺组织中SOD和GSH-Px活性显著降低(P<0.05)。与LPS组比较,PJF干预组肺组织形态改善,ALI评分和肺组织的湿/干重比显著下降(P<0.05),BALF中IL-6、IL-1β和TNF-α水平,以及肺组织中MDA含量、MPO活性及CHOP、GRP78和XBP1蛋白表达水平显著降低(P<0.05),肺组织中SOD和GSH-Px活性显著升高(P<0.05);且高剂量组的效果明显优于低剂量组。结论:PJF对ALI大鼠具有肺保护作用,其机制可能与抑制炎症反应、氧化应激和内质网应激有关。

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。

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。

HPLC法测定泡吴茱萸中柠檬苦素、吴茱萸碱和吴茱萸次碱含量

目的:建立用高效液相色谱法(HPLC)测定泡吴茱萸中柠檬苦素、吴茱萸碱、吴茱萸次碱三种成分的含量。方法:色谱柱为Eclipse XDB C18(4.6×250mm,5μm),流动相:A乙腈,B甲醇,C 0.05%磷酸溶液,各流动相配比为30∶20∶50,流速:1.000mL·min^(-1),检测波长:215nm,柱温:30℃。结果:柠檬苦素、吴茱萸碱、吴茱萸次碱三者依次在0.1303~1.6291μg(r=0.9995)、0.0500~0.6251μg(r=1.0000)、0.0354~0.4429μg(r=0.9999)范围内与其峰面积有良好的线性关系,且三者的平均加样回收率和RSD值分别为100.11%(RSD=0.7%)、100.74%(RSD=1.4%)和99.39%(RSD=1.3%)。结论:该方法操作简单,结果稳定,有着高准确度,适用于泡吴茱萸中柠檬苦素、吴茱萸碱、吴茱萸次碱含量的测定。

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。

Analysis of Volatile Compounds and Identification of Characteristic Aroma Components of <i>Toona sinensis</i>(A. Juss.) Roem. Using GC-MS and GC-O

In this study, volatile compounds present in Toona sinensis (A. Juss.) Roem (TS) were investigated and their characteristic aromatic components were identified using Headspace Solid-phase Microextraction (HS-SPME) followed by Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography-Olfactometry (GC-O). The optimum conditions for extracting the volatiles from TS were achieved with the experimental parameters including the use of a 65 μm polydimethylsiloxane/divinyl benzene (PDMS/DVB) fibre, an extraction temperature of 40℃ and an extraction time of 30 min. Under these conditions, 56 volatile compounds were separated and 53 were identified by GC-MS. Among them, 21 sulfide compounds (42.146%) and 27 terpenes(55.984%) were found to be the major components. The sample was analyzed by GC-O and 26 elutes were sniffed and their sensory descriptions evaluated by an odor panelists. Analysis of the data indicated, two compounds cis and trans isomers of 2-Mercapto-3,4-dimethyl-2,3-dihydrothiophene were major contributors to the characteristic aroma of TS.

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.

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。

Effect of Different Plant Growth Regulators on Callus Induction and in vitro Rapid Propagation of Wild Petunia Juss.

In this study,the seeds of wild Petunia Juss.were used as explants to investigate the optimal condition for tissue culture.Several different kinds and concentrations of growth regulators were adopted to produce more multiple bud clumps,callus or roots in this study.The experiments may provide experimental foundation for the rapid propagation technology and establishment of tissue culture system for wild Petunia Juss.

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.

Toxicity Experiment of Polysaccharides from Dicliptera chinensis( L. ) Juss.

[Objectives] To study the acute toxicity,mutation,and feeding experiment of polysaccharides from Dicliptera chinensis( L.)Juss.,and assess its safety. [Methods]In accordance with Procedures for toxicological Assessment on Food Safety,acute toxicity to mice,micronucleus experiment for bone marrow cell in mice,sperm shape abnormality test in mice,and Ames experiment were carried out,and 30 d feeding experiment was detected for rats. [Results] when the maximum tolerance dose( MTD) of mice was higher than 20 g/kg,at polysaccharides from Dicliptera chinensis( L.) Juss. ≤10 g/kg,there was no statistical difference between the control group and mouse bone marrow cell micronucleus and sperm shape abnormality test group; Ames experiment indicated that the number of revertant colonies of different dose groups of polysaccharides from Dicliptera chinensis( L.) Juss. with and without S9 did not exceed two times of the blank control group; rat 30 d feeding experiment indicated that when polysaccharides from Dicliptera chinensis( L.) Juss. ≤ 3. 2 g/kg,rats grew and developed well,and there was no statistical difference with the control group in blood,multiple organ biochemical indicators and liver tissue. [Conclusions]Polysaccharides from Dicliptera chinensis( L.) Juss. are safe and non-toxic chemical compounds and can be used as health products or drugs.

Chemical constituents and biological activities of Azadirachta indica A. Juss

Azadirachta indica A. Juss is a tropical evergreen plant, belonging to the Meliaceae family. The plant is used as folk medicines for the treatment of fever, neuromuscular pains, skin infections, diabetes and heart disease. Previous researches show that the plant possesses several types of chemical constituents and diverse biological activities. The paper concludes chemical constituents, biological activities and pesticide products of A. indica.

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.

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。