Anti-tumor Effect and Protective Effect on Chemotherapeutic Damage of Water Soluble Extracts from Hedyotis diffusa

Bai-Hua-She-She-Cao Hedyotis diffusa Willd. (Ru-biaceae) is a medicinal herbwidely distributed in northeast Asian countries. In traditional Chinese medicine, it has the effectof 'clearing away heat and toxic material, promoting blood circulation and removing blood stasis'.It is a well known Chinese folk-medicine used for the treatment of appendicitis, sore throat, mumps,acne, sebo-rheic dermatitis and various kinds of tumors, such as tumors of digestive tract,carcinoma of liver. It was reported that the MeOH extract of H. diffusa demonstrated a significantantitumor activity and ursolic acid succeeded in being isolated from the MeOH extract as an activecomponent . Shan BN, et al suggested that the direct aqueous extract of H. diffusa hadimmuno-modulating activity and antitumor activity in vitro through stimulating the immune system tokill or engulf tumor cells. But regarding anti-tumor activity in vivo of water soluble extracts fromH. diffusa, no detail was reported. Therefore, we prepared water soluble extracts (H_1 and H_2)from H. diffusa and evaluated their anti-tumor property in vivo experiments as well as protectiveeffect on chemo-therapeutic damage.

Physiological Effect and Quality Influences of Hedyotis diffusae Extract on Tobacco Infected by TMV

The extract from Hedyotis diffusae was used to treat tobacco plants infected by TMV before and after incidence. The effect of the changes of some physiological and biochemical indices related with the virus-resistance in tobacco leaves on tobacco quality ware determined. The resuits showed that H. diffusae extract could remarkably increase the total phenol content in tobacco leaves ,strength the activity of phenylalanine ammonialyase （ PAL） , improve the membrane permeability and tobacco quality. Prevention treatment was better than therapy treatment. In conclusion, H. diffusae extract could induce plants to produce disease resistance and strength tha antiviral capability of tobacco against TMV.

Phytochemical screening and in vitro bioactivities of the extracts of aerial part of Boerhavia diffusa Linn.

Objective:To investigate the bioactivities of crude n-hexane,ethyl acetate and methanol extracts of aerial part of Boerhavia diffusa Linn.(B.diffusa) and its phytochemical analysis.Methods: the identification of phyloconstituents and assay of antioxidant,thrombolytic,cytotoxic, antimicrobial activities were conducted using specific standard in vitro procedures.Results:The results showed that the plant extracts were a rich source of phytoconstituents.Methanol extract showed higher antioxidant,thrombolytic activity and less cytotoxic activity than those of n-hexane and ethyl acetate extracts of A diffusa.Among the bioactivities,antioxidant activity was the most notable compared to the positive control and thus could be a potential rich source of natural antioxidant.In case of antimicrobial screening,crude extracts of the plant showed remarkable antibacterial activity against tested microorganisms.All the extracts showed significant inhibitory activity against Candida albicuns,at a concentration of 1000 μg/disc.Conclusions:The present findings suggest that,the plant widely available in Bangladesh,could be a prominent source of medicinally important natural compounds.

Anti-tumor research progress of Herba Hedyotidis Diffusae

Cancer,as one of the major public health problems in the world,dramatically harms human health.In recent years,studies have found that Herba Hedyotidis Diffusae not only plays a huge role in anti-infection but also has a strong effect on anti-tumor.Several studies have shown that the components contained in Herba Hedyotidis Diffusae have different levels of antitumor results in multiple pathways.This article summarizes the antitumor elements in Herba Hedyotidis Diffusae,and describes the antitumor mechanism of Herba Hedyotidis Diffusae.Provide new ideas for further clinical use.

Antitumor effects and compatibility mechanism of Hedyotis Diffusa-Sculellaria Barbata herb pair

OBJECTIVE Hedyotis diffusa Willd and Scutellaria barbata D Don are most commonly used herb pair for cancer treatment in traditional Chinese medicine.This study aimed to evaluated the in vitro and in vivo antitumor effects and the compatibility mechanisms of Hedyotis diffusa-Sculellaria barbata herb pair.METHODS Hedyotic diffusa extract,Scutellariae Barbatae extract,and herb pair extract were prepared and the component were detected by UPLC-Q-TOF-MS.The effects of different concentrations of Hedyotic diffusa extract,Scutellariae Barbatae extract,and herb pair extract were detected using MTS assay.The in vivo antitumor effects were evaluated in Panc28 pancreatic cells bearing nude mice model.The compatibility mechanism of Hedyotis diffusa-Sculellaria barbata herb pair were evaluated based on the network pharmacology,using TCMSP,Cytoscape 3.6.1 software,and DAVID 6.8.The anticancer mechanism were further validated in vitro.RESULTS Both the MTS and in vivo results showed that herb pair extract showed more obvious inhibitory effects on cancer cells compared to each individual.A total of 37 active components were selected from Hedyotis diffusa and Sculellaria barbata,33 kinds of active ingredients are involved in their anti-tumor effects.58 cancer-related targets and 66 KEGG pathways were identified.The potential targets for the herb pair might be prostaglandin G/H synthase 2(PTGS2),HSP90,EGFR,72 ku typeⅣcollagenase,PPAR-γ,et al.In vitro validation result showed that compatibility mech⁃anisms was related with HSP90,EGFR related pathways.CONCLUSION The result of the study preliminarily verified the basic anti-tumor pharmacological effects of Hedyotis diffusa-Sculellaria barbata herb pair,and lays a solid foundation for further studies on the anti-tumor mechanism of the herbal pair.

Introduction and Cultivation of Viola diffusa Ging and Its Gardening Application in Hubei

Distribution of Viola diffusa Ging in Hubei was introduced fi rst,then its biological and ecological characters,introduction and cultivation,and reproduction techniques elaborated. In addition,its gardening application in Wuhan was explored.

Cytotoxic, Antimicrobial and Antioxidant Properties of <i>Commelina diffusa</i>Burm. F.

Commelina diffusa Burm. F. is a herbaceous tropical plant with different traditional medicinal uses. Present study is aimed to isolate the bioactive compounds from DCM-Methanol extract of the powdered whole plant of Commelina diffusa and to investigate the cytotoxic, antimicrobial and antioxidant activities of the crude extract and its twelve vacuum liquid chromatographic fraction (CD1-12). Vacuum Liquid Chromatography (VLC) was employed to isolate bioactive metabolites. The physicochemical properties of the isolated compound were examined and the molecular structure was elucidated by NMR spectroscopy.?Cytotoxic, antibacterial, antioxidant activities were evaluated following brine shrimp lethality bioassay, disc diffusion method and DPPH free radical scavenging assay respectively. The isolated compound was identified as steroid (stigmasterol) which had significant cytotoxic effect on vero cell line. The crude extract and its fractions (CD8-CD12) exhibited strong cytotoxicity in brine shrimp lethality bioassay having LC50 values 3.79, 9.19, 29.49, 16.60, 19.36, 44.58 μg/mL respectively. The crude extract showed mild to strong antibacterial activity. Fractions (CD5-CD12) showed mild to strong antibacterial activity against Escherichia coli in comparison with Kanamycin (standard). Strong antioxidant activities were found in crude extract (IC50 value 30.52 μg/ml), CD11 (IC50 value 39.27 μg/ml) and CD12 (IC50 value 19.50 μg/ml). The study suggests Commelina diffusa plant extract to have strong antioxidant and cytotoxic activity which is indicative of presence of compounds with broad spectrum of curative applications. One compound namely stigmasterol has been isolated from the plant.

Effect of Polysaccharides from Hedyotis diffusa on Immune Cells of Small Intestinal Mucosa of Chicks

[Objectives]This study aimed to investigate the effect of polysaccharides from Hedyotis diffusa on the immune cells of the intestinal mucosa of chicks.[Methods]Total 200 seven-day-old AA chicks were evenly and randomly divided into four groups:control group and three treatment groups.The chicks in the control group(group I)were fed basal diet,and those in the treatment groups were fed with basal diet supplemented with 0.4(group II),0.8(group III)and 1.2(group IV)g/kg H.diffusa polysaccharides(OPS),respectively.At the age of 14,28 and 42 d,10 chicks from each of the groups were sacrificed,respectively for counting of intraepithelial lymphocytes(IELs)and lamina propria secretory IgA(SIgA)positive cells in small intestinal mucosa.[Results]At the age of 14 d,the duodenal and jejunal IEL and SIgA positive cell counts in groups II,III and IV were significantly greater than those in group I(P<0.05).At the age of 28 d,the jejunal IEL count of group IV was significantly lower than those of groups I,II and III(P<0.05);and there was no significant difference in IEL or SIgA positive cell count between group I and the other groups(P>0.05).At the age of 42 d,the duodenal and jejunal IEL counts in group II were significantly greater than those in groups I,III and IV(P<0.05),while the differences in duodenal and jejunal IEL count between groups III,IV and I were insignificant(P>0.05).The duodenal SIgA positive cell counts in the treatment groups were insignificantly different from that in group I(P>0.05).The jejunal SIgA positive cell count in group II was significantly greater than those in groups I,III and IV(P<0.05).There was no significant difference in jejunal SIgA positive cell count between groups III and IV and group I(P>0.05).[Conclusions]OPS at 0.4 g/kg can promote the proliferation of immune cells in the small intestine mucosa of 14-42-day-old AA chicks,and it is recommended to be added to the feed.

Effects of Oldenlandia diffusa Polysaccharides on the Structures of Chick Small Intestine Tissues

In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including 50 chicks.The control group(group Ⅰ)was fed with basal diet;and the 3 experimental groups were fed based on the basal diet with the addition of OPS at 0.4 g/kg(group Ⅱ),0.8 g/kg(group Ⅲ),and 1.2 g/kg(group Ⅳ)feed for 1 week,respectively,and then fed the basal diet for 4 weeks.At the ages of 14,28,and 42 d,10 chickens were put into death in the control group and the experimental groups,respectively,and the duodenum and jejunum tissue samples were collected for the analysis of the effects of OPS on the structures of chick intestines.The results showed that at the age of 14 d,the villus lengths of duodenum and jejunum in groups Ⅱ,Ⅲ,and Ⅳ were significantly greater than those in group Ⅰ(P<0.05),and the ratios of duodenal or jejunal villus length to crypt depth in groups Ⅱ and Ⅲ were significantly greater than that in group Ⅰ(P<0.05);at the age of 28 d,the villus length of duodenum in group Ⅱ was significantly greater than that in group Ⅰ(P<0.05);and at the age of 42 d,the ratios of jejunual villus length to crypt depth in groups Ⅱ,Ⅲ and Ⅳ were significantly larger than those of group I(P<0.05).It showed that OPS could promote the small intestine development of AA broilers aged 14 to 42 d.

<i>In Vitro</i>Propagation and Cytological Analysis of <i>Boerhaavia diffusa </i>L—An Important Medicinal Plant Species of Bangladesh

An effective and consistent in vitro propagation technique for Boerhaavia diffusa L. was developed using different types of explants and media compositions. Shoot apex and nodal explants of field grown plants were aseptically cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of auxins (IAA and NAA) and cytokinins (BAP and Kn). The maximum number of multiple shoot buds (6.65 ± 0.09) obtained from nodal explants on MS medium containing 2.0 mg/l BAP + 0.5 mg/l IAA. Multiple shoot buds underwent rapid elongation (4.24 ± 0.06 cm) on MS media fortified with 3.0 mg/l BAP + 0.5 mg/l NAA. Half strength MS media supplemented with 1.0 mg/l IBA + 0.5 mg/l IAA was better for induction and proliferation (6.42 ± 0.07) of roots and 75% of plantlets were successfully acclimatized to ex vitro condition, exhibiting a normal development. Somatic chromosome number of in vivo and in vitro grown plants were confirmed to be 2n = 26. Chromosome length ranged from 1.40 to 2.43 μm in the mother plants and 1.34 to 2.31 μm for in vitro grown plants. The total form percent (TF%) of mother and in vitro grown plants was 42.91% and 41.16%, respectively. The karyotype formula of in vivo grown plants was 2n = 26 = 4Lsm + 6Msm + 2Mm + 14Sm, whereas that of the in vitro grown plants was 2n = 26 = 8Lsm + 4Msm + 2Mm + 12Sm. The frequency of the chromosome having arm more than 2:1 was 0.08 for in vivo grown plants and 0.15 for in vitro raised plants. Thus, according to Stebbins classification (1971) the karyotype of both plants falls into 2A symmetrical type.

Flavonoid-Rich Extract of Oldenlandia diffusa(Willd.)Roxb.Inhibits Gastric Cancer by Activation of Caspase-Dependent Mitochondrial Apoptosis

Objective:To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells,determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species(ROS),and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.Methods:Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin.Bioactive components were characterized by high-performance liquid chromatography.Cell lines MKN-45,AGS,and GES-1 were treated with different concentrations of flavonoids(64,96,128,and 160μg/mL).The effect of flavonoids on cell viability was evaluated by MTT method,and cell nuclear morphology was observed by Hoechst staining.The apoptosis rate and cell cycle phases were measured by flow cytometry,the production of ROS was detected by laser confocal microscope,the mitochondrial membrane potential(MMP)were observed by fluorescence microscope,and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting.MKN-45cells were transplanted into BALB/c nude mice to establish a xenograft tumor model.Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue.The tumor volume and tumor weight were measured,the expression levels of proliferation markers proliferating cell nuclear antigen(PCNA)and Ki-67were detected by immunohistochemistry,and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.Results:Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells,arrested the cell cycle in G1/S phase,induced accumulation of ROS in the process of apoptosis,and altered MMP.In addition,flavonoids increased Apaf-1,Cleaved-Caspase-3,and Bax,and decreased Cyclin A,Cdk2,Bcl-2,Pro-Caspase-9,and Mitochondrial Cytochrome C(P<0.05).The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors.The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group,the expression level of CA72-4 decreased(P<0.05).Conclusion:Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.

3D Microfluidic System for Evaluating Inhibitory Effect of Chinese Herbal Medicine Oldenlandia diffusa on Human Malignant Glioma Invasion Combined with Network Pharmacology Analysis

Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluidic chip culture and the possible mechanism of action of Oldenlandia diffusa Will.(OD).Methods:The comprehensive pharmacodynamic analysis method in this study was based on microfluidic chip 3D cell perfusion culture technology,and the action mechanism of Chinese medicine(CM) on human MG cells was investigated through network pharmacology analysis.First,the components of EEOD were analyzed by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then,cell viability and apoptosis were assessed to determine the optimum concentration of EEOD for invasion experiments,and two-dimensional(2D) migration and invasion abilities of U87 and U251 MG cells were evaluated using scratch wound and Transwell assays.The possible mechanism underlying the effects of EEOD on glioma was analyzed through a network pharmacology approach.Results:Thirty-five compounds of EEOD were detected by UPLC-Q-TOF/MS.EEOD suppressed the viability of MG cells,promoted their apoptosis,and inhibited their migratory and invasive potentials(all P<0.05).Network pharmacology analysis showed that OD inhibited the invasion of MG cells by directly regulating MAPK and Wnt pathways through MAPK,EGFR,MYC,GSK3B,and other targets.The anti-invasion effect of OD was also found to be related to the indirect regulation of microtubule cytoskeleton organization.Conclusion:EEOD could inhibit the invasion of human MG cells,and the anti-invasion mechanism of OD might be regulating MAPK and Wnt signaling pathways and microtubule cytoskeleton organization.

Anti-tumor activity of Hedyotis diffusa Willd. in mice

The whole plant with root ofHedyotis diffusa Willd. （AE） has been demonstrated to have anti-inflammatory, antioxidant, and anti-bacterial activities. In our study, we aim to examine the anti-tumor effect of alcoholic extract of AE in mice implanted with sarcoma S180 cells （SBT mice）. We also compared the immune system function and the life span of SBT mice with that of control mice. We found that AE displayed a significant inhibitory effect on solid tumor growth, as well as increased the life span. At the dose of 10 mg/kg body weight, both tumor weight and volume were decreased significantly. We also measured several immune fimction markers, inlcuding spleen index （SI）, thymus index （TI） and parameters of hematological. We observed no reduce in these markers, indicatine that AE could inhibit tumor ~rowth without affectino immune function in SBT ,nice.

不同配伍比例的半枝莲-白花蛇舌草药对对肝癌细胞增殖活性的影响

目的:研究不同配伍比例的半枝莲-白花蛇舌草药对(简称半白药对,SB-OD)对肝癌细胞株(MHCC97H细胞和Huh7细胞)增殖活性的抑制作用。方法:运用CCK-8法观察不同配伍比例的半白药对(半白1∶1,半白2∶1,半白1∶2)干预MHCC97H和Huh7细胞24 h,分别计算其IC_(50)值;并应用细胞平板克隆形成实验检测细胞克隆形成能力,分析不同配伍比例的半白药对对MHCC97H和Huh7细胞增殖活性的影响。结果:不同配伍比例的半白药对作用MHCC97H和Huh7细胞后的生存活力明显下降(P<0.01),并呈浓度依赖性,其中半白1∶1组作用于MHCC97H和Huh7细胞的IC_(50)值分别为6.69 mg/mL和7.95 mg/mL;半白2∶1组作用于MHCC97H和Huh7细胞的IC_(50)值分别为2.58 mg/mL和3.43 mg/mL;半白1∶2组作用于MHCC97H和Huh7细胞的IC_(50)值分别为15.04 mg/mL和16.60 mg/mL。不同配伍比例的半白药对干预MHCC97H和Huh7细胞后的克隆形成率明显降低(P<0.01)。结论:不同配伍比例的半白药对均能够抑制肝癌MHCC97H和Huh7细胞的增殖活性,2∶1配伍比例的抑制作用最佳,可作为半白药对抗肿瘤研究的实验基础。

基于PI3K/Akt/p53信号通路探讨白花蛇舌草乙酸乙酯萃取物诱导急性髓系白血病细胞凋亡的作用

目的通过评估白花蛇舌草乙酸乙酯萃取物(EAEHDW)对磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/p53信号通路传导调节作用,探讨其抑制急性髓系白血病M2型(AML-M2)细胞株Kasumi-1增殖及诱导凋亡的机制。方法利用乙酸乙酯从白花蛇舌草中萃取制备得到EAEHDW,高效液相色谱串联质谱装置检测样品纯度。设置空白组,将浓度分别为0.02 mg/mL、0.04 mg/mL、0.06 mg/mL、0.08 mg/mL、0.10 mg/mL的EAEHDW加入对数生长期的Kasumi-1细胞株,采用MTT法检测不同浓度EAEHDW作用不同时间后Kasumi-1细胞存活率,采用Annexin V/PI流式细胞术检测不同浓度EAEHDW作用24 h后Kasumi-1细胞凋亡情况,采用Western blot法检测不同浓度EAEHDW(0.02 mg/mL、0.04 mg/mL、0.06 mg/mL)作用24 h后Kasumi-1细胞中含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)家族蛋白及PI3K/Akt/p53信号传导通路蛋白表达情况。结果不同浓度的EAEHDW干预后,细胞存活率较空白组明显降低,细胞凋亡率较空白组明显升高,且细胞存活率随着作用时间的延长和药物浓度的增加而下降越明显,细胞凋亡率随着药物浓度的增加而升高越明显,差异均有统计学意义(P均<0.05);不同浓度的EAEHDW作用于Kasumi-1细胞后,细胞中多聚ADP核糖聚合酶(PARP)、Caspase-3、Caspase-8、Caspase-9、细胞色素C(Cyto-C)、p53蛋白相对表达量均明显升高(P均<0.05),Akt、磷酸化Akt(p-Akt)、鼠双微染色体2(MDM2)、磷酸化MDM2(p-MDM2)蛋白相对表达量均明显降低(P均<0.05)。结论EAEHDW可明显抑制髓系白血病Kasumi-1细胞增殖并诱导其凋亡,其机制可能与影响Caspase家族蛋白表达和抑制PI3K/Akt/p53信号通路导致p53激活有关。

多维度评价电子束辐照灭菌对白花蛇舌草质量的影响

为多维度评价电子束辐照灭菌技术在白花蛇舌草中的应用前景,研究考察了电子束辐照灭菌对白花蛇舌草微生物水平、化学成分含量、指纹图谱、体外抗氧化活性及细胞存活率的影响。结果表明,电子束辐照灭菌对白花蛇舌草中微生物具有明显杀灭作用。吸收剂量2~10 kGy时,白花蛇舌草中芦丁与车叶草苷含量、3种抗氧化活性、细胞存活率较灭菌前均无显著差异(P>0.05),吸收剂量大于10 kGy时,抗氧化活性及十六烷酸甲酯含量降低,建议安全吸收剂量为4~10 kGy。灭菌后白花蛇舌草指纹图谱较对照指纹图谱的相似度均大于0.90。液质联用(LC-MS)确认了指纹图谱中14个共有峰结构,灰色关联度分析(GRA)显示14个共有峰与3种抗氧化活性的关联度(r)均大于0.5,其中与1,1-二苯基-2-三硝基苯肼自由基(DPPH·)反应率超过50%的峰为8、10、11号峰。本研究可为电子束辐照灭菌技术在白花蛇舌草生产中的应用提供科学依据。

清热解毒法在恶性肿瘤治疗中的临床应用

现代中医肿瘤学的癌毒病机理论提出,清热解毒法为治疗恶性肿瘤的抗癌解毒八法之一,可在恶性肿瘤治疗中广泛应用。清热解毒法治疗恶性肿瘤的临床应用需基于辨证、辨病、分期、辨体相结合的综合辨治模式。若肿瘤患者热毒证候明显,辨证应用清热解毒法无疑;若热毒不明显或无证可辨,结合现代肿瘤的病理本质特征,在不违背整体遣方用药原则下,可辨病应用清热解毒法以发挥抗癌祛毒之效。清热解毒法本质属于攻法,其辨证或辨病应用均应以患者正气强弱为考量因素,同时还需结合患者的肿瘤分期、体质因素等。辨证应用强调了清热解毒法运用于肿瘤热毒证的普遍性,辨病应用突破了清热解毒法仅用于宏观热毒证的局限性,分期应用体现了清热解毒法阶段性使用的精准性,辨体应用则体现了清热解毒法应用过程中的个体性。辨证、辨病、分期、辨体4种辨治模式当相互参照、综合运用,方可取得显效。治疗肿瘤常用的清热解毒药物有白花蛇舌草、半枝莲、重楼、天葵子、漏芦、山豆根等。

白花蛇舌草三大类成分的分离纯化及抗肿瘤活性测定

目的从白花蛇舌草中分离纯化总三萜、总多糖、总黄酮,并检测其抗肿瘤活性。方法将白花蛇舌草置于乙醇溶液中冷浸,对其中的三萜类、多糖和黄酮类化合物进行粗提;再通过大孔吸附树脂对粗提物进行处理,获得纯化后的总三萜、总多糖、总黄酮。通过CCK-8法检测白花蛇舌草总三萜、总多糖、总黄酮对A549细胞增殖的影响。结果三萜类化合物浸膏得率为20.60%,精制后总三萜纯度为82.73%;粗多糖得率为41.17%,精制后总多糖纯度为71.11%;黄酮类化合物浸膏得率为0.49%,精制后总黄酮纯度为38.14%。纯化后的白花蛇舌草总三萜、总黄酮和总多糖对A549细胞增殖均有抑制作用,白花蛇舌草总三萜和总黄酮对肺癌细胞A549的抑制作用明显。结论白花蛇舌草总三萜、总黄酮、总多糖均抑制肺癌A549细胞的增殖,为白花蛇舌草总黄酮、总多糖、总三萜的抗肿瘤作用研究提供了一定的参考。

白花蛇舌草提取物通过AMPK/ATG5信号通路对急性胰腺炎大鼠肺损伤的保护作用机制研究

目的:通过5’-单磷酸腺苷活化蛋白激酶(AMPK)/自噬相关蛋白5(ATG5)信号通路,探讨白花蛇舌草提取物减轻急性胰腺炎(AP)大鼠肺损伤的机制。方法:建立AP肺损伤大鼠模型,随机分为模型组、提取物(白花蛇舌草提取物)组、3-MA(自噬抑制剂3-甲基腺嘌呤)组、AICAR(AMPK激活剂)组、提取物+AICAR组,每组15只,另取15只大鼠作为假手术组;ELISA检测血清淀粉酶(AMY)、IL-1β、IL-6、IL-18水平;检测大鼠腹水量及肺湿/干重比(W/D);HE观察胰腺及肺组织病理损伤;Western blot检测溶酶体相关膜蛋白2(LAMP2)、自噬底物p62、IL-1β前体蛋白(pro-IL-1β)、胰蛋白酶原活化肽(TAP)、AMPK、p-AMPK、ATG5、自噬标志物LC3-Ⅱ/Ⅰ、泛素特异性蛋白酶10(UPS10)表达。结果:与假手术组相比,模型组大鼠腹水量、肺W/D、胰腺和肺组织病理损伤评分、血清AMY、IL-1β、IL-6、IL-18水平、胰腺组织p62、TAP、pro-IL-1β表达、肺组织pAMPK/AMPK、ATG5、LC3-Ⅱ/Ⅰ、UPS10、pro-IL-1β表达均升高(P<0.05),胰腺和肺组织LAMP2蛋白表达降低(P<0.05);模型大鼠经白花蛇舌草提取物或自噬抑制剂3-MA干预后,上述指标均得到显著改善(P<0.05),且白花蛇舌草提取物的改善效果优于3-MA(P<0.05);而AMPK激活剂AICAR可削弱白花蛇舌草提取物对AP大鼠肺损伤的改善作用(P<0.05)。结论:白花蛇舌草提取物可通过抑制AMPK/ATG5信号通路减轻炎症反应、降低自噬水平改善AP大鼠肺损伤。

白花蛇舌草多糖对异烟肼致肝损伤的影响及机制

目的探讨白花蛇舌草多糖(HDP)对异烟肼致肝损伤的影响及机制。方法以正常发育的具有肝脏特异性荧光的转基因斑马鱼为对象,分为正常组、模型组(4 mmol/L异烟肼)、HDP低浓度组(4 mmol/L异烟肼+50 mg/mL HDP)和HDP高浓度组(4 mmol/L异烟肼+100 mg/mLHDP),分组处理后观察斑马鱼肝脏荧光面积、荧光强度以及肝组织病理变化。以人肝L02细胞为对象,分为正常组、模型组(4mmol/L异烟肼)、HDP低浓度组(4mmol/L异烟肼+2mg/mLHDP)和HDP高浓度组(4mmol/L异烟肼+4mg/mLHDP),分组处理后检测细胞存活情况,细胞上清液中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平及细胞裂解液中谷胱甘肽(GSH)含量,沉默信息调节因子1(Sirt1)、核转录因子红系2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、醌氧化还原酶1(NQO1)蛋白的表达水平。结果与模型组比较,HDP低、高浓度组斑马鱼肝脏荧光面积和荧光强度(HDP低浓度组除外)均显著增加(P<0.05或P<0.01),肝损伤坏死特征均有不同程度改善。与模型组比较,HDP低、高浓度组L02细胞存活率,GSH含量(HDP低浓度组除外),Sirt1(HDP低浓度组除外)、Nrf2、NQO1、HO-1(HDP低浓度组除外)蛋白表达水平均显著升高(P<0.05或P<0.01);ALT、AST(HDP低浓度组除外)水平均显著降低(P<0.05);存活细胞数量均明显增加,损伤或死亡细胞数量均明显减少。结论HDP对异烟肼诱导的肝损伤具有一定的改善作用,其机制可能与激活Sirt1/Nrf2信号通路、改善线粒体功能、提高抗氧化能力相关。