盐酸氟桂利嗪与鼠神经生长因子治疗脑出血的效果分析

目的研究脑出血患者经盐酸氟桂利嗪与鼠神经生长因子治疗的效果。方法选取自2016年1月—2017年1月宜宾市第二人民医院接受治疗的脑出血142例为研究对象。按治疗方法分成观察组及对照组,每组71例。对照组给予盐酸氟桂利嗪,观察组在对照组基础上加用鼠神经生长因子。观察并比较2组的临床疗效,治疗前后的血肿体积及水肿面积,神经功能缺损评分(NIHSS)及药物不良反应。结果观察组总有效率高于对照组(P<0.05)。2组治疗后的血肿体积及水肿面积小于治疗前,且观察组小于对照组(P<0.05);2组NIHSS评分低于治疗前,且观察组低于对照组(P<0.05)。2组药物不良反应总发生率比较差异无统计学意义(P>0.05)。结论盐酸氟桂利嗪联合鼠神经生长因子治疗脑出血,效果满意,可改善神经功能缺损,安全性较高。

左氧氟沙星辅助抗结核治疗重症结核性脑膜炎临床疗效及对脑脊液指标的影响

目的探讨左氧氟沙星辅助抗结核治疗重症结核性脑膜炎临床疗效及对脑脊液指标的影响。方法我院收治的98例重症结核性脑膜炎患者,根据治疗方案的不同分为对照组48例和观察组50例,患者均完善相关检查,对照组予常规抗结核治疗,观察组在此基础上加用左氧氟沙星辅助治疗,比较两组治疗效果及脑脊液中相关指标水平,不良反应发生情况和采用左氧氟沙星治疗后期脑脊液中左氧氟沙星浓度。结果观察组临床总有效率高于对照组(P<0. 05);治疗后两组脑脊液压力、蛋白质及白细胞值均较治疗前下降,葡萄糖水平较治疗前升高,观察组变化程度均大于对照组(P<0. 05);两组不良反应发生率比较,差异无统计学意义(P>0. 05);观察组采用左氧氟沙星治疗后2、4、6 h时其脑脊液左氧氟沙星浓度分别为(0. 86±0. 32)、(2. 46±0. 48)、(2. 18±0. 28)μg/ml。结论左氧氟沙星辅助抗结核药物治疗重症结核性脑膜炎具有更高的临床有效率,且可更好的改善患者脑脊髓情况,具有一定的安全性,值得作为临床上治疗结核性脑膜炎的首选治疗方案之一。

轮叶马先蒿的环烯醚萜苷成分及其抗补体活性

目的:研究轮叶马先蒿的环烯醚萜苷成分及其抗补体活性。方法:通过溶血实验,以抗补体活性为导向分离手段,对轮叶马先蒿各部位进行抗补体活性测试,采用各种色谱法进行分离,根据理化性质和波谱数据鉴定化合物结构,确定化合物的抗补体活性成分。结果:轮叶马先蒿正丁醇部位抗补体活性较强,从中分离鉴定了12个环烯醚萜苷类化合物,分别为:玉叶金花苷酸甲酯(1)、小米草苷(2)、龙船花苷(3)、桃叶珊瑚苷(4)、玉叶金花苷酸(5)、山栀苷甲酯(6)、京尼平苷(7)、7-去氧栀子新苷(8)、乙基胡桃苷(9)、乙基表桃叶珊瑚苷(10)、7-去氧-8-表番木鳖酸(11)、kankanoside A(12)。化合物5、6、11对经典途径的补体激活具有抑制作用。结论:其中,化合物7、8、12为首次从马先蒿属植物中分离得到。化合物1、3、6、9、10、11为首次从该植物中分离得到。化合物5显示出较强的抗补体活性,50%抑制溶血浓度(CH_(50))为164μg/mL,可作为今后开发天然补体抑制剂的候选化合物。

基于HPLC指纹图谱和网络药理学的祛瘀散结胶囊质量标志物预测

基于指纹图谱和网络药理学预测祛瘀散结胶囊质量标志物。采用HPLC法建立祛瘀散结胶囊指纹图谱,使用中药色谱指纹图谱相似度评价系统进行相似度评价,同时进行化学模式识别分析,筛选批次间差异的主要标志性成分,结合网络药理学对筛选出的标志性成分进行靶点和通路分析。结果表明,建立了14个共有峰的指纹图谱;通过对照品指认了7个共有成分,经化学模式识别,筛选出4个差异性成分,分别为迷迭香酸、三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1);结合网络药理学,认为4种化合物可以作为祛瘀散结胶囊潜在的质量标志物。建立的指纹图谱分析方法简单可行,结合网络药理学得到相关靶点和通路,可作为潜在的药效关联物质,为后期的药效机制研究奠定基础。

基于UPLC-Q TOF MS/MS和HPLC的祛瘀散结胶囊的化学成分分析及其多成分含量测定

目的:应用液质联用法分析祛瘀散结胶囊的化学成分,构建祛瘀散结胶囊中有效成分的含量测定方法。方法:采用超高效液相色谱-三重四极杆飞行时间质谱联用(UPLC-Q TOF MS/MS)技术,色谱柱为Hypersil Gold C_(18)(100 mm×2.1 mm, 1.9μm),以乙腈(A)-0.1%甲酸水(B)为流动相,梯度洗脱,体积流量0.4 mL·min^(-1),柱温40.0℃,质谱数据采集为负离子模式扫描;通过数据库匹配、元素组成和碎片结构分析,鉴定祛瘀散结胶囊中的主要化学成分。另外,采用HPLC,色谱柱为Ultimate?AQ-C_(18)(250 mm×4.6 mm, 5μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,体积流量1 mL·min^(-1),柱温25℃,检测波长203 nm;以外标法计算,测定了11个不同批次祛瘀散结胶囊中的柚皮苷、新橙皮苷、三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1)的含量,并以人参皂苷Rg_(1)为内参物,建立一测多评法。结果:从祛瘀散结胶囊中鉴定出29个化合物;外标法测得柚皮苷、新橙皮苷、三七皂苷R_(1)、人参皂苷Rg_(1)、人参皂苷Rb_(1)的含量分别为0.484~1.097、0.341~0.618、1.685~2.399、5.748~8.386、3.868~5.898 mg·g^(-1),一测多评法测得的柚皮苷、新橙皮苷、三七皂苷R_(1)、人参皂苷Rb_(1)的含量分别为0.516~1.153、0.372~0.667、1.794~2.580、4.373~6.690mg·g^(-1),一测多评法计算值与外标法实测值之间相对误差≤8.9%。结论:UPLC-Q TOF MS/MS法能快速鉴定祛瘀散结胶囊的化学成分;建立的外标法稳定可靠,可用于祛瘀散结胶囊的质量控制;以人参皂苷Rg_(1)为内参物建立的一测多评法有较好的可行性,适用于祛瘀散结胶囊日常生产的含量测定。

密枝圆柏的化学成分及其抗补体、抗氧化活性

目的对密枝圆柏Juniperus convallium进行化学成分和抗补体、抗氧化活性研究。方法采用正相硅胶、ODS-C18、SephadexLH-20以及制备型HPLC等方法进行分离纯化,结合理化性质和波谱数据鉴定化合物结构,采用细胞溶血法测定经典、旁路途径的抗补体活性及作用靶点,DPPH、ABTS、FRAP法研究抗氧化活性。结果从密枝圆柏醋酸乙酯部位分离鉴定了17个化合物,包括黄酮类化合物9个:穗花杉双黄酮(1)、柏木双黄酮(2)、柏木双黄酮葡萄糖苷(3)、柚皮素-7-O-葡萄糖苷(4)、芹菜素(5)、山柰酚-3-O-(6″-O-E-肉桂酰基)-β-D-吡喃葡萄糖苷(6)、槲皮素-3-O-β-D-葡萄糖苷(7)、槲皮素吡喃鼠李糖苷(8)和海波拉亭-7-O-β-D-吡喃葡萄糖苷(9);木脂素类化合物3个:异马尾松脂苷B(10)、异落叶松脂素-2a-O-β-D-葡萄糖苷(11)和异落叶松脂素-3a-O-β-D-葡萄糖苷(12);萜类化合物3个:柳叶二醇(13)、3β-羟基山达海松酸(14)和(1R,3R,4a R,4b S,7R,10a R)-7-乙烯基-1,2,3,4,4a,4b,5,6,7,9,10,10a-十二氢-3-羟基-1,4a,7-三甲基-1-菲甲醇(15);以及大丁苷元(16)和β-谷甾醇(17)。除16外,不同类型化合物对补体系统都显示了一定程度的抑制活性,经典途径50%抑制溶血所需浓度(CH50)为0.05~3.99 mmol/L,旁路途径50%抑制溶血所需浓度(AP50)为0.58~19.13 mmol/L,其中,黄酮类化合物尤其是双黄酮类,是密枝圆柏中重要的抗补体活性成分,并且酚羟基和糖苷基团是影响其抗补体活性的重要因素。仅羟基基团较多的黄酮类(1~3、5~9)和木脂素类(10~12)化合物显示不同程度的抗氧化活性。结论所有化合物均为首次从密枝圆柏中分离得到,黄酮及木脂素类是其抗补体和抗氧化的主要活性成分,且显示出一定的构效关系,为密枝圆柏药效物质基础及质量控制研究提供了依据。

Houttuynia cordata polysaccharide alleviated intestinal injury and modulated intestinal microbiota in H1N1 virus infected mice

Houttuynia cordata polysaccharide(HCP) is extracted from Houttuynia cordata, a key traditional Chinese medicine. The study was to investigate the effects of HCP on intestinal barrier and microbiota in H1N1 virus infected mice. Mice were infected with H1N1 virus and orally administrated HCP at a dosage of 40 mg×kg^(–1)×d^(–1). H1N1 infection caused pulmonary and intestinal injury and gut microbiota imbalance. HCP significantly suppressed the expression of hypoxia inducible factor-1α and decreased mucosubstances in goblet cells, but restored the level of zonula occludens-1 in intestine. HCP also reversed the composition change of intestinal microbiota caused by H1N1 infection, with significantly reduced relative abundances of Vibrio and Bacillus, the pathogenic bacterial genera. Furthermore, HCP rebalanced the gut microbiota and restored the intestinal homeostasis to some degree. The inhibition of inflammation was associated with the reduced level of Toll-like receptors and interleukin-1β in intestine, as well as the increased production of interleukin-10. Oral administration of HCP alleviated lung injury and intestinal dysfunction caused by H1N1 infection. HCP may gain systemic treatment by local acting on intestine and microbiota. This study proved the high-value application of HCP.

Bupleurum polysaccharides ameliorated renal injury in diabetic mice associated with suppression of HMGB1-TLR4 signaling

Bupleurum polysaccharides(BPs)is isolated from Bupleurum smithii var.parvifolium,a key traditional Chinese medicine.The study was to investigate the effects of BPs on diabetic kidney injury.After two intraperitoneal injections of streptozotozin(STZ)100 mg·kg^–1,renal injury in diabetic mice was induced and BPs was orally administrated at dosages of 30 and 60 mg·kg^–1·d^–1.The STZ injected mice developed renal function damage,renal inflammation and fibrosis known as diabetic kidney disease(DKD).BPs significantly reduced serum creatinine level and urinary albumin excretion rate,with the attenuated swelling of kidneys.BPs treatment obviously alleviated the pathological damage of renal tissue.The progression of renal injury in BPs treated mice was inhibited with less expression of type IV collagen(Col IV),fibronectin(FN)andα-smooth muscle actin(α-SMA).The inhibition of inflammation in kidney was associated with the reduced level of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6).BPs administration suppressed the over-expression of toll like receptor 4(TLR4)and high-mobility group box 1(HMGB1)with lowered activity of nuclear factor kappa B(NF-κB)in renal tissue of diabetic mice.Oral administration of BPs effectively prevented the development of renal injury in diabetic mice.This study suggested that the protection provided by BPs might affect through the interruption of HMGB1-TLR4 pathway,leading to the inhibition of renal inflammation and fibrotic process.

Heteroclitins R-S: new dibenzocylooctadiene lignans from Kadsura heteroclita

AIM: To study the dibenzocylooctadiene lignans from the stems of Kadsura heteroclita. METHOD: Chromatographic separations of silica gel and semi-preparative HPLC were used. All of the structures were elucidated on the basis of spectroscopic analysis,including 2D-NMR and HR-MS techniques. RESULTS: Four dibenzocylooctadiene lignans were isolated from K. heteroclita. Their structures were identified as heteroclitin R(1), heteroclitin S(2), gonisin O(3), and schisanlignone A(4). CONCLUSION: Heteroclitin R(1) and heteroclitin S(2) are new natural lignans.

Polysaccharides extracted from the roots of Bupleurum chinense DC modulates macrophage functions

The present study aimed to investigate the effects of polysaccharides extracted from Bupleurum chinense DC(BCPs) on macrophage functions. In the in vivo experiment, 1 m L of 5% sodium thioglycollate was injected into the abdomen of the mice on Day 0 and macrophages were harvested on Day 4. The macrophages were cultured in plates and treated with different concentrations of BCPs and stimulus. Effects of BCPs on macrophage functions were assessed by chemotaxis assay, phagocytosis assay and Enzyme-Linked Immunosorbent Assay(ELISA). Our results showed the enhanced chemotaxis, phagocytosis and secretion of nitric oxide(NO) and inflammatory cytokines by macrophages when treated with BCPs. However, when chemotaxis and phagocytosis were up-regulated by complement components or opsonized particles, BCPs inhibited these effects. Also, the NO production induced by lipopolysaccharides(LPS) was suppressed by BCPs mildly. Moreover, BCPs had an inhibitory effect on the [Ca^(2+)]i elevation of macrophages. These results suggested that BCPs exerted modulatory effects on macrophage functions, which may contribute to developing novel approaches to treating inflammatory diseases.

Flavonoids rather than alkaloids as the diagnostic constituents to distinguish Sophorae Flavescentis Radix from Sophorae Tonkinensis Radix et Rhizoma: an HPLC fingerprint study

Sophorae Flavescentis Radix(Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma(S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora(Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis(HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.

Protective effects of Rabdosia japonica var. glaucocalyx extract on lipopolysaccharide-induced acute lung injury in mice

The present study was designed to evaluate the protective effects of ethanol extracts of Rabdosia japonica var. glaucocalyx(Maxim.) Hara(RJ) on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in mice and the possible underlying mechanisms of action. The mice were orally administrated with RJ extract(16, 32 or 64 mg?kg–1) daily for consecutive7 days before LPS challenge. The ung specimens and the bronchoalveolar lavage fluid(BALF) were collected for histopathological examinations and biochemical analyses. Pretreatment with RJ significantly enhanced superoxide dismutase(SOD) activity and reduced the wet-to-dry weight(W/D) ratio, the levels of nitric oxide(NO) and protein leakage, and myeloperoxidase(MPO) activity in mice with ALI, in a dose-dependent manner. RJ reduced complement deposition and significantly attenuated LPS-induced ALI by reducing productions of inflammatory mediators, such as tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-1β(IL-1β). The results demonstrated that RJ may attenuate LPS-induced ALI via reducing the production of pro-inflammatory mediators, and reducing complement deposition and radicals.

Preparative separation and quantitative determination of two kaurenoic acid isomers in root barks ofAcanthopanax gracilistylus

The kaurenoic acid-type diterpenoids in Acanthopanacis Cortex have been reported to be the major active components. However, the diterpenoids are present as position isomers that exacerbate the challenges in obtaining standards compounds. Little work has been done on the quantitative analysis of the diterpenoids in the herb. In the present study, two diterpenoid isomers ent-16βH,17-isovalerate-kauran-19-oic acid(1) and ent-16βH,17-methyl butanoate-kauran-19-oic acid(2) with high purity were separated by analytical HPLC, followed by recrystallization in acetone. Furthermore, an HPLC-ELSD method was developed and validated for simultaneous determination of 1 and 2 in 9 batches of Acanthopanacis Cortex samples. The HPLC separation and quantification was achieved in 40 min using an Agela Promosil C18 column eluted with a gradient of water and acetonitrile. The calibration curves showed good linearity(r^2 ≥ 0.999 9) within the test ranges. The LOD ranged from 0.407 2 to 0.518 0 mg and LOQ ranged from 1.018 0 to 1.295 0 mg. The precisions(%RSD) were within 1.47% for the two isomers. The recovery of the assay was in the range of 98.78%-99.11% with RSD values less than 2.76%. It is the first time to establish a quantitative HPLC method for the analysis of the bioactive kaurenoic acid isomers in the herb.

Polymyxin B as an inhibitor of lipopolysaccharides contamination of herb crude polysaccharides in mononuclear cells

Lipopolysaccharides(LPS) contamination in herbal crude polysaccharides is inevitable. The present study was performed to explore the effect of polymyxin B on abolishing the influence of LPS contamination in mononuclear cells. LPS was pretreated with polymyxin B sulfate(PB) at different concentrations for 1, 5 or 24 h, and then used to stimulate RAW264.7 and mouse peritoneal macrophages(MPMs). The nitric oxide(NO) and tumor necrosis factor-α(TNF-α) in cell culture supernatant, as the indications of cell response, were assayed. Bupleurum chinensis polysaccharides(BCPs) with trace amount contamination of LPS was treated with PB. 30 μg·mL^(–1) of PB, treating LPS(10 and 1000 ng·mL^(–1) in stimulating RAW264.7 and MPMs respectively) at 37 ℃ for 24 h, successfully abolished the stimulating effect of LPS on the cells. When the cells were stimulated with LPS, BCPs further promoted NO production. However, pretreated with PB, BCPs showed a suppression of NO production in MPMs and no change in RAW264.7. In the in vitro experiments, LPS contamination in polysaccharide might bring a great interference in assessing the activity of drug. Pretreatment with PB(30 μg·mL^(–1)) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination(10 and 1 000 ng·mL^(–1)).