Antioxidant activities, metal contents, total phenolics and flavonoids of seven Morchella species

Seven Morchella species were analyzed for their antioxidant activities in different test systems namely β-carotene/linoleic acid, DPPH, reducing power, chelating effect and scavenging effect (%) on the stable ABTS⁺, in addition to their heavy metals, total phenolic and flavonoid contents. In β-carotene/linoleic acid system, the most active mushrooms were M. esculenta var. umbrina and M. angusticeps. In the case of DPPH, methanol extract of M. conica showed high antioxidant activity. The reducing power of the methanol extracts of mushrooms increased with concentration. Chelating capacity of the extracts was also increased with the concentration. On the other hand, in 40 μg ml⁻¹ concentration, methanol extract of M. conica, exhibited the highest radical scavenging activity (78.66 ± 2.07%) when reacted with the ABTS⁺ radical. Amounts of seven elements (Cu, Mn, Co, Zn, Fe, Ca, and Mg) and five heavy metals (Ni, Pb, Cd, Cr, and Al) were also determined in all species. M. conica was found to have the highest phenolic content among the samples. Flavonoid content of M. rotunda was also found superior (0.59 ± 0.01 μg QEs/mg extract).     

灵芝孢子粉多糖Lzps-1的化学研究及其总多糖的抗肿瘤活性

目的研究微波软化灵芝孢子粉中的多糖组分及其抗肿瘤活性。方法用水提取微波软化灵芝孢子粉总多糖，总多糖经分级沉淀得到多糖组分Lzps-C，再采用DEAE-cellulose和Sephadex G-50柱色谱进行分离纯化，用化学和光谱方法分析其结构。结果从微波软化灵芝孢子粉的水提物中分得一个多糖Lzps-1，其平均分子量为8 000，为葡聚糖。微波软化灵芝孢子粉的水提物得到的总多糖Lzps对小鼠Lewis肺癌、小鼠S₁₈₀肉瘤有较好的抑制作用，并能明显提高荷Lewis肺癌小鼠NK活性。结论Lzps-1是首次从灵芝孢子粉中分离得到。Lzps具有抗癌活性。     

Antioxidant and antimicrobial activities of branches extracts of five Juniperus species from Turkey

Context: Several Juniperus species (Cupressaceae) are utilized in folk medicine in the treatment of infections and skin diseases.

Objective: This work was designed to evaluate the antioxidant and antimicrobial potential of methanol and water branches extracts of Juniperus species from Turkey: Juniperus communis L. var. communis (Jcc), Juniperus communis L. var. saxatilis Pall. (Jcs), Juniperus drupacea Labill. (Jd), Juniperus oxycedrus L. subsp. oxycedrus (Joo), Juniperus oxycedrus L. subsp. macrocarpa (Sibth. & Sm.) Ball. (Jom).

Materials and methods: Total phenolics, total flavonoids and condensed tannins were spectrophotometrically determined. The antioxidant properties were examined using different in vitro systems. The toxicity was assayed by Artemia salina lethality test. The antimicrobial potential against bacteria and yeasts was evaluated using minimum inhibitory concentration and minimum bactericidal concentration (MIC/MBC) measurements. The effect on bacteria biofilms was tested by microtiter plate assay.

Results: Both in the DPPH (1,1-diphenyl-2-picrylhydrazyl) and TBA (thiobarbituric acid) test Jom resulted the most active (IC₅₀?=?0.034?±?0.002?mg/mL and 0.287?±?0.166 µg/mL). Joo exhibited the highest reducing power (1.78?±?0.04 ASE/mL) and Fe²⁺ chelating activity (IC₅₀?=?0.537?±?0.006?mg/mL). A positive correlation between primary antioxidant activity and phenolic content was found. The extracts were potentially non-toxic against Artemia salina. They showed the best antimicrobial (MIC?=?4.88-30.10 µg/mL) and anti-biofilm activity (60–84%) against S. aureus.

Discussion and conclusion: The results give a scientific basis to the traditional utilization of these Juniperus species, also demonstrating their potential as sources of natural antioxidant and antimicrobial compounds.     

Anti-tumor and immunoregulatory activities of Ganoderma lucidum and its possible mechanisms

Ganoderma lucidum (G lucidum) is a medicinal fungus with a variety of biological activities. It has long been used as a folk remedy for promotion of health and longevity in China and other oriental countries. The most attractive character of this kind of medicinal fungus is its immunomodulatory and anti-tumor activities. Large numbers of studies have shown that G lucidum modulate many components of the immune system such as the antigen-presenting cells, NK cells, T and B lymphocytes, The water extract and the polysaccharides fraction of G lucidum exhibited significant anti-tumor effect in several tumor-bearing animals mainly through its immunoenhancing activity. Recent studies also showed that the alcohol extract or the triterpene fraction of G lucidum possessed anti-tumor effect, which seemed to be related to the cytotoxic activity against tumor cells directly. Preliminary study indicated that antiangiogenic effect may be involved antitumor activity of G lucidum.     

Antioxidant,anticholinesterase and antibacterial activities of Stachys guyoniana and Mentha aquatica

Context: Stachys guyoniana Noë ex. Batt. and Mentha aquatica L. are two Algerian Lamiaceae used in folk medicine.

Objective: To investigate their antioxidant, anticholinesterase and antibacterial activities.

Material and methods: n-Butanol (BESG), ethyl acetate (EESG) and chloroform (CESG) extracts of S. guyoniana and methanol (MEMA) and chloroform (CEMA) aerial part extracts of M. aquatica and methanol (MERMA) and acetone (AERMA) roots extracts of M. aquatica were evaluated for their antioxidant activity by the β-carotene-linoleic acid, DPPH^? and ABTS^?+?scavenging, CUPRAC and metal chelating assays. The anticholinesterase activity was tested against AChE and BChE. The antibacterial activity was assessed by MICs determination against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella heidelberg, Klebsiella pneumoniae, Enterobacter aerogenes and Morganella morganii strains.

Results: In the β-carotene test, the CESG (IC₅₀: 2.3?±?1.27?μg/mL) exhibited the highest activity. The BESG was the best scavenger of DPPH^? (IC₅₀: 2.91?±?0.14?μg/mL). In the ABTS test, AERMA was the most active (IC₅₀: 4.21?±?0.28?μg/mL). However, with the CUPRAC, the BESG exhibited the best activity (A_0.50: 0.15?±?0.05?μg/mL) and was active in metal chelating assay with 48% inhibition at 100?μg/mL. The BESG was the best AChE inhibitor (IC₅₀: 5.78?±?0.01?μg/mL) however, the AERMA showed the highest BChE inhibitory activity (IC₅₀: 19.23?±?1.42?μg/mL). The tested extracts exhibited a good antibacterial activity.

Conclusion: This study demonstrated good antioxidant, anticholinesterase and antibacterial potential of S. guyoniana and M. aquatica, which fits in well with their use in folk medicine.     

灵芝对高血压伴高血脂和脑血栓后遗症患者血液流变学的观察

灵芝片110mg,qid，服药2周，对高血压伴高血脂、脑血栓后遗症和冠心病等高血粘度病人33例进行血液流变学观察。结果表明，灵芝片有改善血液流变性、降低血粘度作用，因而对心脏血管疾病有减少血栓形成的效果。此外，多数病人服用灵芝后有降压、镇静、催眠和改善头痛、眼花等症状的作用。     

Antioxidant and hepatoprotective activities of five eggplant varieties

Eggplant is consumed throughout the world and varies in fruit color, shape, and size. In this study, five varieties of eggplant (purple colored moderate size, white-green colored moderate size, long green, green striped moderate size and pale-green colored small size, respectively, called SM1–SM5) were evaluated for total phenolic and flavonoid content, antioxidant activity and hepatoprotection against cytotoxicity of tert-butyl hydroperoxide (t-BuOOH) in human hepatoma cell lines, HepG2. Total phenolic content found in methanol extracts of SM1–SM5 ranged from 739.36 ± 1.59 to 1116.13 ± 7.30 gallic acid equivalents mg/100 g extract and total flavonoid content from 1991.29 ± 6.32 to 3954.20 ± 6.06 catechin equivalents mg/100 g extract. SM1 and SM2 which contained high total phenolic and flavonoid had better antioxidant activities than the other varieties. Pretreatment of HepG2 cells with 50 and 100 μg/mL of SM1–SM5 significantly increased the viability (p < 0.05) of t-BuOOH-exposed HepG2 cells by 14.49 ± 1.14% to 44.95 ± 2.72%. The antioxidant activities of the eggplant were correlated with the total amounts of phenolic and flavonoid (r = 0.5310−0.7961). Significant correlation was found between hepatoprotective activities and total phenolic/flavonoid content (r = 0.6371–0.8842) and antioxidant activities (r = 0.5846–0.9588), indicating the contribution of the phenolic antioxidant present in eggplant to its hepatoprotective effect on t-BuOOH-induced toxicity.     

不同原料栽培的灵芝子实体形态结构及活性多糖肽成分的研究

目的观察不同原料栽培的草栽灵芝、鹿角灵芝与段木灵芝子实体形态与构造,分析3种灵芝活性成分多糖含量,提取率及红外光谱识别。方法采用体视显微镜切片观察子实体内部构造。蒽酮-硫酸法测定多糖,lowry法测肽含量。结果草栽和段木栽培灵芝形态正常。控制栽培条件可产生鹿角灵芝,其菌柄顶瑞无子实层分化,未见到菌管和担子及担孢子的结构。菌草灵芝和段木灵芝高分子部位多糖提取率分别为0.94%、0.55%和0.78%,以菌草灵芝为最高;肽含量以段木灵芝为最高。结论栽培条件对灵芝子实体形态与构造有一定的影响,但对活性成分的红外光谱分析变化不大。     

不同制备工艺灵芝孢子粉抗血栓、改善心脏功能作用研究

目的 采用斑马鱼模型,比较研究破壁灵芝孢子粉（wall-broken Ganoderma lucidum spore powder,BGLSP）和去壁灵芝孢子粉（wall-removedGanoderma lucidum spore powder,RGLSP）对血栓形成预防作用及心脏功能改善作用。方法 首先采用紫外可见分光光度法分别测定BGLSP、RGLSP中总多糖和总三萜的含量。其次,在野生型AB品系斑马鱼中进行药物毒性试验,确定灵芝孢子粉的最大耐受浓度。然后,分别用苯肼、花生四烯酸以及普纳替尼诱发斑马鱼血栓模型,并用BGLSP、RGLSP及阳性药干预。建模完成后,用邻联茴香胺进行染色,并用NIS-Elements DTM图像处理软件进行图像分析,计算斑马鱼心脏红细胞染色强度及预防血栓作用。此外,用维拉帕米诱发斑马鱼心衰模型,测定斑马鱼心脏扩大面积、静脉淤血面积、心输出量以及血流速度,定量评价灵芝孢子粉对维拉帕米诱发的斑马鱼心衰的心脏功能改善作用。结果 RGLSP所含多糖和三萜类化合物的含量显著高于BGLSP （P<0.001）,分别提高了11倍和5倍。苯肼、花生四烯酸和普纳替尼诱导后,斑马鱼心脏红细胞染色强度显著增高（P<0.01或P<0.001）。与模型对照组相比,BGLSP高剂量组显著抑制苯肼和普纳替尼诱发的斑马鱼血栓形成（P<0.05）,并且能够显著提高心衰斑马鱼心脏扩大改善率和心输出量增加率（P<0.05或P<0.01）;RGLSP低、中剂量组对苯肼、花生四烯酸和普纳替尼诱发斑马鱼血栓形成,及对维拉帕米诱发斑马鱼心衰均具有显著保护作用（P<0.05或P<0.01或P<0.001）,并且保护作用强于BGLSP。结论 BGLSP和RGLSP均具有显著的抗血栓及抗心衰作用,而RGLSP的药效作用更明显,可能与其较高的有效成分含量有关。     

Antioxidative activities and the total phenolic contents of tonic Chinese Medicinal Herbs

Chinese medicated diet is an everyday practice in China. In this study, 16 commonly used soup making tonic Chinese medicinal herbs were selected for antioxidative capacities by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and ferric reducing antioxidant power (FRAP), and the total phenolic contents of these herbal extracts were measured by the Folin-Ciocalteu method. It confirmed that drinking tonic soups could supplement total antioxidants intake. Amongst the tested herbal extracts, extracts of Canarium album Raeusch., Flos caryophylli and Fructus amomi were found to have the highest antioxidative activities in both DPPH and FRAP assays. Their antioxidative activities were comparable to ascorbic acid and butylated hydroxytoluene. Thus, these herbs are safe and inexpensive sources of natural antioxidants. A significant relationship between the antioxidative effects and total phenolic contents were found, indicating phenolic compounds are the major contributor of antioxidative capacities of these herbs. In addition, a strong correlation between DPPH assay and FRAP assay implied that antioxidants in these herbs were capable of scavenging free radicals and reducing oxidants.     

Antioxidant and antibacterial activities of the leaf essential oils of Himalayan Lauraceae species

The leaf essential oils from seven Himalayan Lauraceae species viz. Neolitsea pallens, Lindera pulcherrima, Dodecadenia grandiflora, Persea duthiei, Persea odoratissima, Persea gamblei and Phoebe lanceolata exhibited potent antioxidant and antibacterial activities. The in vitro antioxidant activity was assessed by using β-carotene bleaching assay, reducing power, DPPH radical scavenging and inhibition of lipid peroxidation methods. The oils of D. grandiflora and L. pulcherrima showed a potent free radical scavenging activity as evidenced by low IC₅₀ value for DPPH radical (0.032 mg/ml and 0.087 mg/ml, respectively) and inhibition of lipid peroxidation (in between IC₅₀ = 0.44 mg/ml and IC₅₀ = 0.74 mg/ml, respectively). The oils were tested against three Gram negative (Escherichia coli, Salmonella enterica enterica and Pasturella multocida) and one Gram positive (Staphylococcus aureus) bacteria at different concentrations using disc diffusion and tube dilution methods. The inhibition zones (IZ) and MIC values for bacterial strains were in the range of 8.7–22.0 mm and 3.90–31.25 μl/ml, respectively.     

Insights into cholinesterase inhibitory and antioxidant activities of five Juniperus species

In vitro acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory and antioxidant activities of the aqueous and ethanol extracts of the leaves, ripe fruits, and unripe fruits of Juniperus communis ssp. nana, Juniperus oxycedrus ssp. oxycedrus, Juniperus sabina, Juniperus foetidissima, and Juniperus excelsa were investigated in the present study. Cholinesterase inhibition of the extracts was screened using ELISA microplate reader. Antioxidant activity of the extracts was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenging, ferrous ion-chelating, and ferric-reducing antioxidant power (FRAP) assays. Total phenol and flavonoid contents of the extracts were determined spectrophotometrically. The extracts had low or no inhibition towards AChE, whereas the leaf aqueous extract of J. foetidissima showed the highest BChE inhibition (93.94 ± 0.01%). The leaf extracts usually exerted higher antioxidant activity. We herein describe the first study on anticholinesterase and antioxidant activity by the methods of ferrous ion-chelating, superoxide radical scavenging, and ferric-reducing antioxidant power (FRAP) assays of the mentioned Juniperus species.     

Antioxidant activities and contents of flavonoids and phenolic acids of Talinum triangulare extracts and their immunomodulatory effects

In this study, leaves and stems of Talinum triangulare were sequentially extracted with phosphate buffer solution to obtain PTL and PTS (phosphate buffered extracts of T. triangulare leaves and stems), with 75% ethanol to obtained ETL and ETS (ethanol extracts of T. triangulare leaves and stems), or with 90°C boiling water to obtain WTL and WTS (water extracts of T. triangulare leaves and stems). We investigated the antioxidant activities of various T. triangulare extracts, analyzed the extracts’ stimulations on human mononuclear cell (MNC) growth and secretion of cytokines (interleukin-1 beta, interferon-γ, and tumor necrosis factor-α) and nitric oxide, and then assayed their subsequent inhibitions on human leukemic U937 cell growth. Results indicated that extracts of T. triangulare showed significant antioxidant activities. Among these extracts, WTS showed the highest stimulatory effect on human MNC growth. The secretion levels of interleukin-1 beta, interferon-γ, and tumor necrosis factor-α in the conditioned medium, wherein human MNC was treated with 500 μg/mL WTS for 72 hours, were 1275, 859, and 2222 pg/mL, respectively. All conditioned media obtained from human MNCs cultured with various T. triangulare extracts showed significant inhibition against U937 cell growth of over 40%. These results suggest that T. triangulare extracts may be used in health foods for their immunomodulatory potential.     

Compounds from the pods of Astragalus armatus with antioxidant,anticholinesterase, antibacterial and phagocytic activities

Context: The phytochemical study and biological activities of Astragalus armatus Willd. subsp. numidicus (Fabaceae) pods, an endemic shrub of Maghreb, are reported.

Objective: This study isolates the secondary metabolites and determines the bioactivities of Astragalus armatus pods.

Materials and methods: The chloroform, ethyl acetate and n-butanol extracts of hydro-ethanolic extracts were studied. Antioxidant activity was investigated using DPPH and ABTS radical scavenging, CUPRAC and ferrous chelating assays at concentrations ranging from 3 to 200?μg/mL. Anticholinesterase activity was determined against acetylcholinesterase and butyrylcholinesterase enzymes at 50, 100 and 200?μg/mL. Antibacterial activity was performed according to minimum inhibitory concentration (MIC) method. Carbon clearance method in albino mice was used for the phagocytic activity at concentrations 50, 70 and 100?mg/kg body weight. Spectroscopic techniques were used to elucidate the compounds.

Results: Ethyl acetate extract afforded a flavonoid (1) while the n-butanol extract gave four flavonoids (2–5), a cyclitol (6) and a cycloartane-type saponin (7). The ethyl acetate extract exhibited highest antioxidant activity in DPPH (IC₅₀: 67.90?±?0.57?μg/mL), ABTS (IC₅₀: 11.30?±?0.09?μg/mL) and CUPRAC (A_0.50: 50.60?±?0.9?μg/mL) assays. The chloroform extract exhibited the best antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, each with 80?μg/mL MIC values. The n-butanol extract enhanced phagocytic activity.

Discussion and conclusion: Isorhamnetin (1), isorhamnetin-3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranoside (2), isorhamnetin-3-O-β-d-apiofuranosyl-(1 → 2)-[α-l-rhamnopyranosyl-(1 → 6)]-β-d-galactopyranoside (3), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-galactopyranoside (4), kaempferol-3-O-(2,6-di-O-α-l-rhamnopyranosyl)-β-d-glucopyranoside (5), pinitol (6) and cyclomacroside D (7) were isolated whereas 1, 2, 6 and 7 are reported for the first time from A. armatus.     

Antioxidant and hepatoprotective activities of the oil fractions from wild carrot (Daucus carota ssp. carota)

Abstract

Context: Wild carrot, Daucus carota L. ssp. carota (Apiacae), is widely distributed throughout the world and has various uses in traditional medicine in Lebanon.

Objective: The present study aimed to fractionate and analyze the chemical composition of the Daucus carota oil extract (DCOE) fractions and to evaluate their antioxidant and hepatoprotective properties in vitro and in vivo.

Materials and methods: DCOE was chromatographed on silica gel column to produce four fractions: pentane (F1), 50:50 pentane:diethyl ether (F2), diethyl ether (F3), and 93:7 chloroform: methanol (F4). Qualitative and quantitative analyses of oil fractions were performed by GC-MS and HPLC techniques. The in vitro antioxidant properties were assessed using DPPH, FIC, and ferric-reducing antioxidant power (FRAP) assays. The hepatoprotective property was determined by examining the levels of serum markers (alanine transaminase (ALT) and aspartate transaminase (AST)) and hepatic antioxidant (superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST)) enzymes in CCl₄-intoxicated mice pretreated with intraperitoenal 50, 100, or 200?mg/kg b.w. of the oil fractions for 5?d.

Results: GCMS analysis of F2 revealed the presence of 2-himachalen-6-ol (61.4%) which is reported for the first time in Daucus carota species. F3 and F4 were rich in phenolics and flavonoids and demonstrated significant DPPH activity (IC₅₀?=?0.29 and 0.38?mg/ml, respectively) and high FRAP values (225.11 and 437.59?µmol FeSO₄/g, respectively). The sesquiterpene-rich fraction F1 had the highest FIC ability (IC₅₀?=?0.28?mg/ml). Pretreatment with F1 and F4 reversed the CCl₄-induced decrease in SOD, CAT, and GST levels and reduced significantly hepatic damage.

Discussion and conclusion: The current results suggested that wild carrot oil fractions exhibited a unique chemical composition and possessed significant antioxidant activities as well as hepatoprotective effects against CCl₄-induced hepatotoxicity.     

野山杏总黄酮对高血脂大鼠降血脂及抗氧化作用的研究

目的测定野山杏果肉中的总黄酮含量,研究野山杏总黄酮(total flavonoids from wild apricot,TFWA)对高血脂大鼠的降血脂作用和抑制肝脏脂质过氧化的能力。方法采用紫外分光光度法,测定野山杏总黄酮的含量;在建立大鼠高脂血症模型的基础上,每天灌胃不同剂量的TFWA,连续预防给药8周,观察TFWA对高血脂大鼠血清中三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)等血脂水平和肝脏中脂质过氧化产物丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)的活性等抗氧化能力的影响。结果①野山杏总黄酮(TFWA)的含量为269.7mg·g-1;②TFWA的高低剂量组较高脂模型组可明显降低肝脏系数(P<0.001)、TC的水平(P<0.001)和TG的水平(P<0.01,P<0.05),TFWA的高剂量对LDL-C水平较高脂模型组可明显降低(P<0.01),对HDL-C的水平可明显升高(P<0.01);③TFWA的高低剂量组较高脂模型组显著降低肝脏中的MDA含量(P<0.01),TFWA的高剂量组较高脂模型组明显升高肝脏中SOD和GSH-Px的含量。结论 TFWA可降低高血脂大鼠的血脂,可减少肝脏损伤,抑制肝脏脂质过氧化过程,并且具有一定的剂量依赖性。     

灵芝多糖微乳诱导结直肠癌肿瘤相关巨噬细胞M1极化及其联合PD-1抑制剂协同抗肿瘤研究

目的:验证灵芝多糖微乳诱导结直肠癌肿瘤相关巨噬细胞(TAMs) M1极化能力,以及联合PD-1抑制剂协同抗肿瘤的可行性。方法:采用"一步成乳法"制备灵芝多糖微乳(GLP-M),并对其进行理化性质表征。借助RAW264.7细胞考察GLP-M诱导巨噬细胞极化的能力;通过Transwell小室法考察GLP-M处理巨噬细胞后对肿瘤细胞增殖、凋亡的影响,以及GLP-M处理肿瘤细胞后对巨噬细胞极化的影响;考察GLP-M诱导巨噬细胞产生活性氧(ROS)行为。建立荷MC38鼠源肠癌细胞的皮下移植瘤模型,以生理盐水和灵芝多糖为阴性对照和剂型对照组,设置GLP-M、PD-1抑制剂以及GLP-M+PD-1联合给药组;PD-1抑制剂每3天尾静脉注射1次,给药2次,其余制剂每天灌胃1次,给药6次。考察治疗期间肿瘤生长曲线、体质量变化、生存时间、病理切片等。结果:GLP-M粒径为(58.3±2.1) nm,Zeta电位为(-23.5±1.8) mV,多分散系数(PDI)为0.221±0.003,包封率为(82.5±3.2)%,TEM图显示GLP-M形态圆整、分散均匀。GLP-M干预IL-4诱导的RAW264.7细胞后CD86的表达量升高(33.2±1.9)%,ROS水平升高(39.4±2.5)%,差异均具有统计学意义。在RAW264.7与MC38细胞共培养模型中,GLP-M可以显著提高肿瘤细胞的增殖抑制率和凋亡率,促进巨噬细胞的CD86表达。体内抗肿瘤实验显示:GLP-M、PD-1以及GLP-M+PD-1的肿瘤抑制率分别为(44.3±4.2)%、(51.3±2.7)%和(63.7±4.3)%。GLP-M+PD-1组45 d生存率最高(25%),且该组肿瘤切片结果显示坏死明显,细胞增殖被显著抑制。结论:GLP-M可能通过干预TAMs M1极化的机制提高与PD-1抑制剂等免疫治疗协同抗结直肠癌治疗效能。     

Sensitive and selective liquid chromatography-tandem mass spectrometry method for the determination of five ganoderic acids in Ganoderma lucidum and its related species

The present paper describes a novel, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of ganoderic acids C(2), B, A, H, D in Ganoderma lucidum and its related species. Ganoderma samples were prepared using simple ultrasonic extraction. Chromatographic separation was carried out on an Agilent Zorbax XDB C(18) column (250 mm × 4.6 mm i.d., 5μm) with an isocratic mobile phase consisting of acetonitrile, water and formic acid (42:58:0.5, v/v/v). Mass spectrometric detection was achieved by a triple-quadrupole mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) interface operating in negative and positive ionization mode via a single within-run polarity switching. Quantitation of five ganoderic acids was performed using selective reaction monitoring (SRM) mode. The intra- and inter-day precision was less than 6.2% and the accuracy ranged from 90.0% to 105.7%. The limit of quantification (LOQ) was 20.0-40.0 ng/mL and the limit of detection (LOD) was 3.0-25.0 ng/mL. With this method, low levels of ganoderic acids in the fruiting bodies of Ganoderma sinense and Ganoderma applanatum were accurately quantified for the first time. Importantly, the method allows unequivocal quantification of the five ganoderic acids in the spores and fruiting bodies of Ganoderma lucidum, whereas the previously published methods have lacked the capability. The method presented will be a powerful tool for quality control of Ganoderma lucidum and its related species.