白芍总苷在正常大鼠体内的组织分布研究

探讨白芍总苷在正常大鼠体内的组织分布特点,为预测其药理作用及不良反应提供依据.正常大鼠按2.82 g/kg灌胃给予TGP药液后1、3、6h取心、肝、脾、肺、肾、胃、小肠、大肠等组织,各组织匀浆后,将匀浆液制成冻干粉,HPLC法测定冻干粉中芍药苷和芍药内酯苷浓度,计算各组织中两者浓度.结果显示1h各组织中均能测到芍药苷和芍药内酯苷,3h除胃和小肠外,其他各组织中两者浓度均达到最大值,小肠、胃、大肠及肾、脾、肝中浓度较高,6h小肠、大肠、胃中浓度较高,其他各组织中浓度较低.说明灌胃TGP后组织分布迅速且广泛,胃、小肠、大肠及肾、脾、肝是主要分布器官,容易在胃肠蓄积,其他组织中蓄积较少,为进一步研究白芍总苷的药理作用及作用机理提供了指导,同时为白芍归经理论提供了一定的现代科学依据.     

白芍总苷对心肌缺血再灌注大鼠内质网应激因子CHOP、GRP78、GRP94的表达及凋亡的影响

目的:研究白芍总苷(TGP)对心肌缺血再灌注(I/R)大鼠内质网应激因子CCAAT/增强子结合蛋白的同源蛋白(CHOP)、葡萄糖调节蛋白78(GRP78)、葡萄糖调节蛋白94(GRP94)表达及凋亡的影响。方法:选择健康清洁级SD大鼠75只,根据随机数字表法分成5组,每组15只,分别记为假手术组、I/R组、50 mg/kg TGP组、100 mg/kg TGP组以及200 mg/kg TGP组。检测并对比各组大鼠CHOP、GRP78、GRP94水平,对比分析各组大鼠心肌I/R指标、梗死面积率以及心肌细胞的凋亡率。结果:I/R组和TGP各组的CHOP、GRP78及GRP94水平均明显高于假手术组,且TGP各组较I/R组明显更低(均P0.05)。100 mg/kg和200 mg/kg TGP组的CHOP、GRP78及GRP94水平均明显低于50 mg/kg TGP组,且200 mg/kg TGP组较100 mg/kg TGP组明显更低(均P0.05)。I/R组和TGP各组缺血30 min的T波改变、再灌注120 min的T波改变及LVEDP水平均明显高于假手术组,LVSP、+dp/dtmax及-dp/dtmax水平均明显低于假手术组(均P0.05)。与I/R组相比,TGP组缺血30 min的T波改变、再灌注120 min的T波改变及LVEDP水平呈剂量依赖型下降,而LVSP、+dp/dtmax及-dp/dtmax水平呈剂量依赖型上升(均P0.05)。I/R组和TGP各组的梗死面积率和心肌细胞的凋亡率均明显高于假手术组(均P0.05)。与I/R组相比,TGP组的梗死面积率和心肌细胞的凋亡率水平呈剂量依赖型下降(均P0.05)。结论:应用TGP能够明显降低MIRI大鼠内质网应激因子CHOP、GRP78、GRP94的表达,调节心肌缺血和再灌注相关标志物或临床参数的水平,显著减少心肌缺血和再灌注所致的心肌梗死面积率及细胞凋亡率。     

不同品质类型小麦谷蛋白聚合体含量及亚基组成的初步研究

以6个不同品质类型小麦品种为试验材料,对其面粉总蛋白含量(FP％)、谷蛋白总聚合体含量(TGP％)、大聚合体含量(GMP％)进行了测定和比较,并利用多层浓缩胶SDS—PAGE对不同品种小麦面粉大聚合体亚基组成进行了初步分析。结果表明：(1)面包和面条型小麦面粉谷蛋白总聚体含量明显高于饼干型小麦;(2)分子量约为32—43kD和14kD的亚基主要是组成麦谷蛋白大聚合体。     

HPLC特征图谱结合含量测定的白芍及其炮制品的质量对比研究

建立白芍、炒白芍、酒白芍、硫熏白芍HPLC特征图谱,并结合多成分含量测定,为白芍、炒白芍、酒白芍和硫熏白芍的质量控制提供参考。采用Intersustain C₁₈(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.1%醋酸水溶液,流速为每分钟1 mL,梯度洗脱,检测波长为230 nm,柱温为30℃,进样量为10μL。14批白芍、炒白芍、酒白芍和硫熏白芍的特征图谱,标定了6个共有峰,并均被指认,分别为没食子酸、儿茶素、芍药内酯苷、芍药苷、1,2,3,4,6-五没食子酰葡萄糖和苯甲酰芍药苷,而硫熏白芍标定7个共有峰,峰7为白芍硫熏后产生;且各色谱谱峰有较好的分离,但不同炮制品特征图谱存在一定差异;含量测定结果显示,白芍炒制、酒制及硫熏后,6种成分均有不同程度的变化;借助中药色谱指纹图谱相似度评价系统和SIMCA-P13.0软件对14批白芍、炒白芍、酒白芍和硫熏白芍进行相似度和正交偏最小二乘判别(OPLS-DA)分析,所建立的白芍和炮制品及硫熏品的质量评价方法稳定性、重复性好,可用于白芍、炒白芍、酒白芍和硫熏白芍的质量控制和评价。     

HS-SPME-GC-MS分析柴胡、白芍配伍前后挥发性组分变化规律

为了探讨柴胡-白芍药对配伍的意义,通过单因素考察确定最佳萃取条件,采用顶空固相微萃取法(HS-SPME)与气相色谱-质谱(GC-MS)联用,测定柴胡、白芍及其药对挥发性成分,通过面积归一法确定各成分百分含量,并进行主成分分析(PCA)。结果表明：柴胡、白芍及其药对共有的挥发性成分有11种,但共有成分的含量有所不同。三者的PCA综合得分具有明显的差异,药对挥发性成分的PCA综合得分最高。柴胡-白芍药对的挥发性成分大部分来自于柴胡,柴胡、白芍配伍后有新的活性成分产生,从挥发性角度分析,柴胡、白芍配伍后具有一定增效减毒作用。     

白芍总苷治疗慢性荨麻疹的疗效观察

目的:观察白芍总苷治疗慢性荨麻疹的疗效及不良反应.方法:采用随机对照临床试验将60例符合纳入标准的患者随机分为三组,分别为白芍总苷胶囊1.8 g/d加依匹斯汀10 mg/d组(A组),依匹斯汀加复方甘草酸苷150 mg/d组(B组)和单用依匹斯汀10 mg/d组(C组).疗程均为12周.治疗前后检查患者的血、尿、粪常规及肝、肾功,同时观察记录患者的不良反应.结果:A组有效率为90.0％,B组有效率为85.0％,C组有效率为55.0％.A组和B组有效率差异无统计学意义(P＞0.05);A、B组疗效高于对照组,差异有统计学意义(P＜0.05).三组的不良反应均较少.结论:白芍总苷联合依匹斯汀治疗慢性荨麻疹疗效显著且不良反应少,是慢性荨麻疹可供选择的有效治疗方法.     

白芍水提物的抗炎作用及作用机制研究

目的与白芍总苷进行比较,研究白芍水提物的抗炎作用,探讨其抗炎作用的机制。方法采用二甲苯致小鼠耳片肿胀模型和蛋清致大鼠足跖肿胀模型,观察白芍水提物和白芍总苷对急性炎症的抗炎作用。采用琼脂致小鼠肉芽肿模型,观察白芍水提物和白芍总苷对慢性炎症的抗炎作用;通过测定琼脂致小鼠肉芽肿模型血清前列腺素E2(PGE2)水平,蛋清致大鼠足跖肿胀模型肿胀足的一氧化氮(NO)、丙二醛(MDA)及PGE2含量,探究其抗炎作用的部分机制。结果白芍水提物4 g·kg-1、白芍总苷180 mg·kg-1均可显著抑制二甲苯致小鼠耳片肿胀和琼脂肉芽肿(P<0.05),降低琼脂致小鼠肉芽肿模型血清PGE2水平(P<0.05)。白芍水提物2.8 g·kg-1、白芍总苷120 mg·kg-1均可显著抑制蛋清致大鼠足跖肿胀(P<0.05),降低肿胀足的NO、MDA及PGE2含量(P<0.05);白芍水提物和白芍总苷对急、慢性炎症的抑制作用差异无统计学意义(P>0.05)。结论白芍水提物具有一定的抗炎作用,其抗炎机制可能与减少血液和局部组织PGE2水平,减少局部组织NO、MDA含量有关。     

白芍传统品种原植物遗传多样性及亲缘关系分析

为探讨白芍原植物各居群间的遗传多样性及亲缘关系,本实验应用RAPD技术,使用40条随机引物对10个白芍原植物居群的叶片总DNA进行PCR扩增。结果表明：白芍10个原植物居群间具有较丰富的遗传多样性,多态率达63．以％。由结合线L＝0．7975可将10个居群划分为6组,原植物为芍药(Paeonia lactiflora Pall.)的6个居群中有4个居群集中在第3组,显示了种内遗传关系的相似性,其中川白芍(芍药)花色有变,同时雄蕊有部分辨化的现象,与野生种(韩城芍药)有较远的遗传关系;山东荷泽居群雄蕊全部瓣化,种质可能来自于观赏芍药突变体,在遗传关系上与芍药的其他居群有一定的距离。原植物为毛果芍药[P.lactiflora var.trichocarpa (Bunge) Stern]的4个居群中,杭白芍粉红花居群和白花居群独立成一组,与形态分类的结果一致。芍药及其变种毛果芍药的遗传关系非常近,但是有交叉。因此形态分类和遗传关系应综合考虑,作为种下分类单位的确定也应慎重。     

基于HPLC指纹图谱的亳白芍不同炮制品标准汤剂7个成分含量测定

建立亳白芍不同炮制品标准汤剂HPLC指纹图谱,其中生白芍、炒白芍和酒白芍标准汤剂分别标定16、14和13个共有峰,同时测定亳白芍不同炮制品标准汤剂中7种化学成分(没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酸、1,2,3,4,6-五没食子酰葡萄糖和苯甲酰芍药苷)含量,经炒制后标准汤剂中芍药内酯苷和苯甲酸含量升高,氧化芍药苷、芍药苷含量降低,酒白芍标准汤剂中芍药苷含量升高,苯甲酸含量降低,并对含量测定结果进行聚类分析,同一批亳白芍及其炮制品可聚为一类,但聚类距离缩小后生品和炮制品各聚为一类。建立的HPLC指纹图谱和含量测定方法具有良好的重现性,且简便快速,二者结合可直观反映出亳白芍炒制、酒制后的变化差异。     

相同基源的赤芍和白芍中芍药苷含量的比较

通过采用HPLC法比较了中江产芍药的根部不同加工品-赤芍和白芍中芍药苷的含量,以期为研究二者功效差异的具体原因奠定基础.结果表明,赤芍和白芍的化学成份及其含量均存在一定差异,大、中、小三个规格的赤芍和白芍中芍药苷含量分别2.97%、2.94%、2.98%和1.86%、1.82%、1.91%;白芍中芍药苷含量低于赤芍,芍药苷含量与规格无明显关系,化学成份及其含量的差异可能是二者功效差异的具体原因之一.     

Gene expression profile of human mesenchymal stem cells during osteogenesis in three-dimensional thermoreversible gelation polymer

This study attempted to characterize the ability of thermoreversible gelation polymer (TGP) to induce differentiation of human mesenchymal stem cells (hMSC) into osteoblasts. Using a long oligo microarray system consisting of 3760 genes, we compared the expression profiles of the cells in 2-dimensional (2D) culture, 3D culture in collagen gel, and 3D culture in TGP with or without osteogenic induction. Compared to 2D culture, the gene expression profile of hMSC showed almost the same pattern in TGP without osteogenic induction, but 72% of genes (2701/3760) were up-regulated in collagen gel. With osteogenic induction, hMSC showed higher ALP activity and osteocalcin production in TGP as compared to 2D culture. Moreover, up-regulation and down-regulation of osteogenic genes were augmented in 3D culture in TGP as compared to 2D culture. As TGP is chemically synthesized and completely free from pathogen such as prion in bovine spongiform encephalopathy, these results suggest that TGP could be applied clinically to induce osteogenic differentiation of hMSC.     

Purification of thymocyte growth peptide (TGP) from sheep thymus. Relationship to FTS/thymulin

Thymocyte growth peptide (TGP) initiates DNA synthesis in immature thymocytes and has previously been characterized as an acidic peptide isolated from calf thymus. We now report the isolation of TGP from sheep thymus and show it to be a nonapeptide with a large N-terminal blocking moiety characterized by high UV absorbance. The amino acid composition is identical to FTS, consisting of 2 Gly, 2 Ser, 2 Glx, 1 Ala, 1 Lys, 1 Asx. In contrast to FTS, TGP is acidic with an apparent isoelectric point of 4.2 and a high UV absorbance at 270–280 nm. Reverse phase chromatography of TGP at an acidic pH results in a change of the molecule and the appearance of two new compounds TGP-A and TGP-B, both with less than 50% of the original TGP activity. Full activity could be restored by the addition of ZnCl₂ to TGP-A. Both TGP-A and B have some amino acid composition and high UV absorbance as native TGP. We propose that TGP consists of a non-peptide moiety bound to the N-terminal of the nonapeptide Glu-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn and that the active molecule is stabilized by Zn²⁺.     

Mechanistic insight into the attenuation of gouty inflammation by Taiwanese green propolis via inhibition of the NLRP3 inflammasome

Dysregulation of NACHT, LRR, and PYD domains-containing protein 3 (NLRP3) inflammasome is involved in many chronic inflammatory diseases, including gouty arthritis. Activation of the NLRP3 inflammasome requires priming and activation signals: the priming signal controls the expression of NLRP3 and interleukin (IL)-1β precursor (proIL-1β), while the activation signal leads to the assembly of the NLRP3 inflammasome and to caspase-1 activation. Here, we reported the effects of the alcoholic extract of Taiwanese green propolis (TGP) on the NLRP3 inflammasome in vitro and in vivo. TGP inhibited proIL-1β expression by reducing nuclear factor kappa B activation and reactive oxygen species (ROS) production in lipopolysaccharide-activated macrophages. Additionally, TGP also suppressed the activation signal by reducing mitochondrial damage, ROS production, lysosomal rupture, c-Jun N-terminal kinases 1/2 phosphorylation and apoptosis-associated speck-like protein oligomerization. Furthermore, we found that TGP inhibited the NLRP3 inflammasome partially via autophagy induction. In the in vivo mouse model of uric acid crystal-induced peritonitis, TGP attenuated the peritoneal recruitment of neutrophils, and the levels of IL-1β, active caspase-1, IL-6 and monocyte chemoattractant protein-1 in lavage fluids. As a proof of principle, in this study, we purified a known compound, propolin G, from TGP and identified this compound as a potential inhibitor of the NLRP3 inflammasome. Our results indicated that TGP might be useful for ameliorating gouty inflammation via inhibition of the NLRP3 inflammasome.     

Study on the purification and chemical compositions of tea glycoprotein

In this paper, improvement in the method for purifying glycoprotein from green tea (Camellia sinensis) was described; some properties and chemical compositions of tea glycoprotein (TGP) were determined by HPGPC, FT-IR, GC–MS technologies. Compared to existing methods, a more compatible method for purifying TGP was proposed. This method was faster, simpler, more effective and easier to be extended to the industrial production than the method that used in our previous work. The molecular weight of TGP was 126,513 Da using HPGPC. GC–MS analysis of TGP showed that TGP was composed of seven kinds of monosaccharides, namely ribose, rhamnose, arabinose, xylose, mannose, glucose, galactose in molar ratios of 1.71:5.88:13.70:1.99:1.00:1.84:33.75. Eighteen amino acids were identified in TGP by amino acid analysis. The FT-IR spectrum of the TGP revealed also typical characteristics of polysaccharides, protein and uronic acid.     

Effects of tobacco glycoprotein (TGP) on the immune system. I. TGP is a T-independent B cell mitogen for murine lymphoid cells

It has been shown that tobacco glycoprotein (TGP), a polyphenol-rich glycoprotein antigen purified from cured tobacco leaves, is mitogenic for lymphoid cells in the spleen, peripheral blood, and bone marrow, but not for thymus cells. The proliferative response is not reduced by treatment of spleen cells or peripheral blood lymphocytes with anti-Thy-1.2 and complement, and spleen cells from the congenitally athymic (nu/nu) CD-1 proliferate as vigorously in response to TGP as do spleen cells from their heterozygous nu/+ littermates. In addition, TGP induces differentiation of mouse spleen cells into antibody-secreting cells, the majority of which secrete IgM, and the remainder mainly IgG and a few IgA. The differentiation into antibody-secreting cells induced by TGP occurs with spleen cells from nu/nu mice. It is concluded that TGP is a T-independent B cell mitogen for mouse lymphoid cells. On the basis of the ability of spleen cells from the LPS-nonresponder C3H/HEJ mice to respond to TGP with proliferation and differentiation into antibody-secreting cells, it is concluded that the effects of TGP are distinct from those of LPS and cannot be due to contamination of the TGP preparation with LPS.     

Effects of tobacco glycoprotein (TGP) on the immune system. II. TGP stimulates the proliferation of human T cells and the differentiation of human B cells into Ig secreting cells

We have been studying the effects of tobacco glycoprotein (TGP), a polyphenol-rich glycoprotein isolated from cured tobacco leaves, on the immune system. We have shown previously that mice immunized with TGP produce preferentially antibodies of the IgE isotype and that TGP is a T cell-independent B cell mitogen for mice, which stimulates B cell proliferation and B cell differentiation into Ig-secreting cells. We report herein that TGP stimulates a significant increase in [3H]TdR incorporation by human PBL and by human cord blood lymphocytes. The magnitude of the proliferative response of PBL to TGP does not correlate with the donor's titer of IgE antibodies to TGP, as assayed by a wheal and flare response after an i.d. injection of TGP, neither does it correlate with the donor's smoking history. [3H]TdR uptake is not observed before day 5 of culture, and the response peaks between days 5 and 10 of culture. Analysis of the cellular basis for the proliferative response suggests that T cells are proliferating. Two-parameter analysis by flow cytometry shows that CD3+, CD4+, and CD8+ cells are in the S + G2 + M phases, but not Ig-bearing cells or monocytes. A significant increase in HLA-DR (Ia)-bearing cells is observed on cells in all of the cell cycle phases. This increase coincides with cells entering the S phase. No increase is observed in the expression of the IL-2-R as assayed by the anti-Tac antibody. TGP also stimulates human PBL to differentiate and to produce Ig of the IgM, IgG, and IgA isotypes, without stimulating a detectable B cell proliferative response. The proliferative response of PBL is clearly due to TGP and not to contamination with LPS, because by the limulus amebocyte assay the TGP preparation contains less than 2% LPS, which could not account for the stimulation observed.     

Paeoniflorin modulates gut microbial production of indole-3-lactate and epithelial autophagy to alleviate colitis in mice

BackgroundTotal glucosides of peony (TGP), extracted from the root and rhizome of Paeonia lactiflora Pall, has well-confirmed immunomodulatory efficacy in the clinic. However, the mechanism and active ingredients remain largely unclear.Hypothesis/PurposeOur previous study revealed a low systemic exposure but predominant gut distribution of TGP components. The aim of this study was to investigate involvement of the gut microbiota in the immunoregulatory effects and identify the active component.MethodsMice received 3% DSS to establish a model of colitis. The treatment group received TGP or single paeoniflorin (PF) or albiflorin (AF). Body weight, colon length, inflammatory and histological changes were assessed. Gut microbiota structure was profiled by 16s rRNA sequencing. Antibiotic treatment and fecal transplantation were used to explore the involvement of gut microbiota. Metabolomic assay of host and microbial metabolites in colon was performed.ResultsTGP improved colonic injury and gut microbial dysbiosis in colitis mice, and PF was responsible for the protective effects. Fecal microbiota transfer from TGP-treated mice conferred resilience to colitis, while antibiotic treatment abrogated the protective effects. Both TGP and PF decreased colonic indole-3-lactate (ILA), a microbial tryptophan metabolite. ILA was further identified as an inhibitor of epithelial autophagy and ILA supplementation compromised the benefits of TGP.ConclusionOur findings suggest that TGP acts in part through a gut microbiota-ILA-epithelial autophagy axis to alleviate colitis.     

IL-1, IL-6, and PDGF mRNA expression in alveolar cells following stimulation with a tobacco-derived antigen.

To test the hypothesis that inflammatory cytokine production might be an early event in the development of the disease associated with smoking, we used alveolar cells from healthy nonsmokers stimulated with TGP as a model system. TGP, a phenol-rich glycoprotein which is present in tobacco leaves and cigarette smoke condensate, activates the immune system. It stimulates polyclonal B cell differentiation, induces primarily an IgE response, and activates human leukocytes to produce IL-1. Using in situ nucleic acid hybridization we show that the steady-state levels of IL-1 alpha, IL-1 beta, IL-6, platelet-derived growth factor (PDGF)-A, and PDGF-B mRNAs are consistently elevated in the alveolar cells of all donors following TGP stimulation. The kinetics of mRNA expression suggest that IL-1 alpha and IL-1 beta mRNAs are independently regulated in alveolar cells, while the regulation of PDGF-A and PDGF-B mRNA seems to be similar. The activated cells also synthesize elevated levels of IL-1 and IL-6. These findings lend support to the suggestion that some clinical consequences of smoking might be initiated and enhanced by the production of inflammatory cytokines. Moreover, IL-6 could also activate a polyclonal B cell response, which could lead to the synthesis of autoantibodies and thus cause immune-mediated tissue injury.     

Identification of a mammalian growth factor as a ribofolate peptide

Thymocyte growth peptide (TGP) promotes DNA synthesis of immature thymocytes. TGP has been purified from sheep, human and calf thymus and recently characterized as an N-terminally blocked nonapeptide. Evidence is presented here that the blocking moiety consists of a formylpteroyl group bound to the N-terminal glutamyl residue of the nonapeptide. The pterin part of the TGP molecule has a ribityl substituent in analogy with riboflavin, which explains the pronounced hydrophilic property of TGP in contrast to unsubstituted and unconjugated folates. The compound can be classified as a ribofolate peptide, a novel class of growth factor. Zn²⁺ counteracts degradation of the molecule and is required for full biological activity; mass spectrometric data confirm that native TGP contains zinc.     

Transgenerational plasticity and climate change experiments: Where do we go from here?

Phenotypic plasticity, both within and across generations, is an important mechanism that organisms use to cope with rapid climate change. While an increasing number of studies show that plasticity across generations (transgenerational plasticity or TGP) may occur, we have limited understanding of key aspects of TGP, such as the environmental conditions that may promote it, its relationship to within‐generation plasticity (WGP) and its role in evolutionary potential. In this review, we consider how the detection of TGP in climate change experiments is affected by the predictability of environmental variation, as well as the timing and magnitude of environmental change cues applied. We also discuss the need to design experiments that are able to distinguish TGP from selection and TGP from WGP in multigenerational experiments. We conclude by suggesting future research directions that build on the knowledge to date and admit the limitations that exist, which will depend on the way environmental change is simulated and the type of experimental design used. Such an approach will open up this burgeoning area of research to a wider variety of organisms and allow better predictive capacity of the role of TGP in the response of organisms to future climate change.