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Identification of Baicalin as an Immunoregulatory Compound by Controlling TH17 Cell Differentiation  [PDF]
Ji Yang,Xue Yang,Yiwei Chu,Ming Li
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0017164
Abstract: TH17 cells have been implicated in a growing list of inflammatory disorders. Antagonism of TH17 cells can be used for the treatment of inflammatory injury. Currently, very little is known about the natural compound controlling the differentiation of TH17 cells. Here, we showed that Baicalin, a compound isolated from a Chinese herb, inhibited TH17 cell differentiation both in vitro and in vivo. Baicalin might inhibit newly generated TH17 cells via reducing RORγt expression, and together with up-regulating Foxp3 expression to suppress RORγt-mediated IL-17 expression in established TH17 cells. In vivo treatment with Baicalin could inhibit TH17 cell differentiation, restrain TH17 cells infiltration into kidney, and protect MRL/lpr mice against nephritis. Our findings not only demonstrate that Baicalin could control TH17 cell differentiation but also suggest that Baicalin might be a promising therapeutic agent for the treatment of TH17 cells-mediated inflammatory diseases.
Optimal combination of baicalin, icariin and Astragalus saponin Ⅰ from a Chinese herbal compound Biminne
Wei-yi GONG
Zhong Xi Yi Jie He Xue Bao , 2010,
Abstract: Objective: To study the optimal combined ratio of baicalin, icariin and Astragalus saponin Ⅰ from a Chinese herbal compound Biminne. Methods: Firstly, a mouse model of allergic rhinitis was established by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide gel suspension, and the effective dose range of baicalin, icariin and Astragalus saponin Ⅰ was detected by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt method. Secondly, 10 groups of combinations of baicalin, icariin and Astragalus saponin Ⅰ assembled by U*10(108) form were employed to determine the optimal combination by means of analyzing of the inhibitory effect on the splenocyte proliferation. Finally, the effects of each effective ingredient and the optimal combination were compared by observing the splenocyte proliferation, the contents of interleukin-4 (IL-4), IL-5, interferon-gamma (IFN-γ) in supernatant of the splenocyte cultures and the ratio of IL-4 to IFN-γ in order to verify the result. Results: Baicalin or icariin at concentrations ranging from 2 to 10 μmol/L, and Astragalus saponin Ⅰ from 1 to 10 μmol/L effectively suppressed the splenocyte proliferation. When the proportion of baicalin, icariin and Astragalus saponin Ⅰ was 1∶2.14∶2.65, the inhibitory effect was most remarkable. Further research confirmed the rationality of the optimal combination.Conclusion: An optimal combination of the major effective ingredients from Chinese herbal compound Biminne most effectively suppresses the proliferation of splenocytes from sensitized mice and regulates the cytokine secreting.
The Metabolism of Baicalin in Rat and the Biological Activities of the Metabolites
Yi Wang,Jingyu Yang,Xian Li,Jinhui Wang
Evidence-Based Complementary and Alternative Medicine , 2012, DOI: 10.1155/2012/404529
Abstract: Baicalin is one of the major bioactive constituents of Scutellariae Radix, but the biotransformation of it is poorly understood. In this paper, the metabolism of baicalin in rat was studied. Nine metabolites including one new compound were isolated and identified structurally. The plausible scheme for the biotransformation pathways of baicalin in the rats was deduced. And the main metabolites were evaluated for their antioxidation and anti-inflammation biological activities for the first time.
CRABP2 Promotes Myoblast Differentiation and Is Modulated by the Transcription Factors MyoD and Sp1 in C2C12 Cells  [PDF]
Jing Yuan, Zhonglin Tang, Shulin Yang, Kui Li
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0055479
Abstract: Cellular retinoic acid binding protein 2 (CRABP2), a member of a family of specific carrier proteins for Vitamin A, belongs to a family of small cytosolic lipid binding proteins. Our previous study suggested that CRABP2 was involved in skeletal muscle development; however, the molecular function and regulatory mechanism of CRABP2 in myogenesis remained unclear. In this study, we found that the expression of the CRABP2 gene was upregulated during C2C12 differentiation. An over-expression assay revealed that CRABP2 promotes myogenic transformation by regulating the cell cycle during C2C12 differentiation. The region from ?459 to ?4 bp was identified as the core promoter and contains a TATA box, a GC box and binding sites for the transcription factors MyoD and Sp1. Over-expression, site-directed mutagenesis and EMSA assays indicated that the transcription factors MyoD and Sp1 regulate CRABP2 expression and promote myoblast differentiation in C2C12 cells.
Transcriptional regulatory network for sexual differentiation in fission yeast
Juan Mata, Anna Wilbrey, Jürg B?hler
Genome Biology , 2007, DOI: 10.1186/gb-2007-8-10-r217
Abstract: We used DNA microarrays to investigate meiotic gene regulation by examining transcriptomes after genetic perturbations (gene deletion and/or overexpression) of rep1, mei4, atf21 and atf31, which encode known transcription factors controlling sexual differentiation. This analysis reveals target genes at a genome-wide scale and uncovers combinatorial control by Atf21p and Atf31p. We also studied two transcription factors not previously implicated in sexual differentiation whose meiotic induction depended on Mei4p: Rsv2p induces stress-related genes during spore formation, while Rsv1p represses glucose-metabolism genes. Our data further reveal negative feedback interactions: both Rep1p and Mei4p not only activate specific gene expression waves (early and middle genes, respectively) but are also required for repression of genes induced in the previous waves (Ste11p-dependent and early genes, respectively).These data give insight into regulatory principles controlling the extensive gene expression program driving sexual differentiation and highlight sophisticated interactions and combinatorial control among transcription factors. Besides triggering simultaneous expression of gene waves, transcription factors also repress genes in the previous wave and induce other factors that in turn regulate a subsequent wave. These dependencies ensure an ordered and timely succession of transcriptional waves during cellular differentiation.Meiosis and the formation of specialized gametes are fundamental processes of sexual reproduction. Diploid cells of the fission yeast Schizosaccharomyces pombe undergo two meiotic nuclear divisions to produce four stress-resistant spores in response to environmental stimuli [1,2]. This sexual differentiation is accompanied and driven by an extensive gene expression program, during which a large proportion of all genes are either induced or repressed [3-5]. We have previously classified the genes that are up-regulated at least four-fold into four maj
cAMP promotes differentiation of rodent neuronal progenitor cells  [cached]
Guilherme Lepski,Cinthia Elim Jannes,Guido Nikkhah
Stem Cell Studies , 2011, DOI: 10.4081/scs.2011.e9
Abstract: Numerous studies have described neuronal differentiation of neural progenitor cells derived from fetal tissue in vitro, but the biochemical mechanisms underlying this process remain largely unknown. In the present study, the role of cAMP in promoting functional maturation of neuronal progenitor cells (NPCs) from the subventricular zone (SVZ) of rodent fetal brain was investigated. NPCs were extracted from telencephalic vesicles of E14 rat embryos and then expanded in medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). A mature neuronal fate was induced by: i) withdrawal of growth factors (basal condition); ii) addition of brain-derived neurotrophic factor (BDNF); or iii) addition of isobutylmethylxantine (IBMX). Whole cell patch clamping assessed electrophysiological properties. Immunocyto - chemistry for MAP2 confirmed neuronal differentiation. Quantification of neuronal cells, and determination of their electrophysiological properties, was performed in the cited experimental groups. IBMX significantly enhanced the yield of MAP2-positive neurons in the culture system (3.7-fold increase). Application of a one-week differentiation protocol under IBMX induced functional maturity. MAP2-positive cells presented large Na- and K- voltagedependent currents, fired bursts of action potentials, and exhibited spontaneous synaptic activity. Further, IBMX proved more effective than BDNF for promoting neuronal maturation, leading to higher evoked peak currents and current densities, as well as greater yields of cells firing action potentials and presenting active synaptic contacts. Our data indicates the importance of cAMP-dependent mechanisms for maturation of neuronal progenitor cells in vitro. This knowledge can enable future manipulation of neurogenesis in vivo in order to promote the intrinsic regenerative capacity of the central nervous system.
Maternal Baicalin Treatment Increases Fetal Lung Surfactant Phospholipids in Rats
Chung-Ming Chen,Leng-Fang Wang,Kur-Ta Cheng
Evidence-Based Complementary and Alternative Medicine , 2011, DOI: 10.1093/ecam/nep073
Abstract: Baicalin is a flavonoid compound purified from the medicinal plant Scutellaria baicalensis Georgi and has been reported to stimulate surfactant protein (SP)-A gene expression in human lung epithelial cell lines (H441). The aims of this study were to determine whether maternal baicalin treatment could increase lung surfactant production and induce lung maturation in fetal rats. This study was performed with timed pregnant Sprague-Dawley rats. One-day baicalin group mothers were injected intraperitoneally with baicalin (5 mg/kg/day) on Day 18 of gestation. Two-day baicalin group mothers were injected intraperitoneally with baicalin (5 mg/kg/day) on Days 17 and 18 of gestation. Control group mothers were injected with vehicle alone on Day 18 of gestation. On Day 19 of gestation, fetuses were delivered by cesarean section. Maternal treatment with 2-day baicalin significantly increased saturated phospholipid when compared with control group and total phospholipid in fetal lung tissue when compared with control and 1-day baicalin groups. Antenatal treatment with 2-day baicalin significantly increased maternal growth hormone when compared with control group. Fetal lung SP-A mRNA expression and maternal serum corticosterone levels were comparable among the three experimental groups. Maternal baicalin treatment increases pulmonary surfactant phospholipids of fetal rat lungs and the improvement was associated with increased maternal serum growth hormone. These results suggest that antenatal baicalin treatment might accelerate fetal rat lung maturation.
Modulation of P1 and EGF Expression by Baicalin  [PDF]
Yanli Meng,Jinhai Huo,Weihong Lu,Xin Wang,Junwei Zhang,Weiming Wang
International Journal of Molecular Sciences , 2013, DOI: 10.3390/ijms14010146
Abstract: Mycoplasma pneumoniae ( M. pneumoniae) is increasingly recognized as a major cause of acute respiratory tract infections. Today, macrolides are used in the primary treatment of M. pneumoniae infection. However, with the increasing prevalence of strains resistant to macrolides, as well as reports of toxicity and adverse side effects, it is necessary to develop an alternative therapeutic agent. A compound recipe — Qinbaiqingfei pellets (Qinbai) — have already been approved in China as the first effective traditional Chinese medicine to be used against M. pneumoniae. Herein, we characterize the mechanism by which Qinbai interacts with M. pneumoniae and lung epithelial cells. The fact that Baicalin is the key component of Qingbai leads us to believe its study is important to elucidating the mechanism of the action of Qinbai. In this study, we describe the complex impact of Baicalin on the adhesin protein P1 of M. pneumoniae and on the expression of epidermal growth factor (EGF) in BALB/c mice and A549 cells infected with M. pneumonia. We draw the conclusion that Baicalin not only cured M. pneumoniae infection by inhibiting P1 expression, but also enhanced the repair of lung epithelial cells by upregulating EGF. Finally, we demonstrate that Baicalin plays a role in Qinbai treatment.
Microgravity Promotes Differentiation and Meiotic Entry of Postnatal Mouse Male Germ Cells  [PDF]
Manuela Pellegrini,Sara Di Siena,Giuseppina Claps,Silvia Di Cesare,Susanna Dolci,Pellegrino Rossi,Raffaele Geremia,Paola Grimaldi
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0009064
Abstract: A critical step of spermatogenesis is the entry of mitotic spermatogonia into meiosis. Progresses on these topics are hampered by the lack of an in vitro culture system allowing mouse spermatogonia differentiation and entry into meiosis. Previous studies have shown that mouse pachytene spermatocytes cultured in simulated microgravity (SM) undergo a spontaneous meiotic progression. Here we report that mouse mitotic spermatogonia cultured under SM with a rotary cell culture system (RCCS) enter into meiosis in the absence of any added exogenous factor or contact with somatic cells. We found that isolated Kit-positive spermatogonia under the RCCS condition enter into the prophase of the first meiotic division (leptotene stage), as monitored by chromosomal organization of the synaptonemal complex 3 protein (Scp3) and up-regulation of several pro-meiotic genes. SM was found to activate the phosphatidyl inositol 3 kinase (PI3K) pathway and to induce in Kit-positive spermatogonia the last round of DNA replication, typical of the preleptotene stage. A PI3K inhibitor abolished Scp3 induction and meiotic entry stimulated by RCCS conditions. A positive effect of SM on germ cell differentiation was also observed in undifferentiated (Kit-negative) spermatogonia, in which RCCS conditions stimulate the expression of Kit and Stra8. In conclusion, SM is an artificial environmental condition which promotes postnatal male germ cell differentiation and might provide a tool to study the molecular mechanisms underlying the switch from mitosis to meiosis in mammals.
Aryl Hydrocarbon Receptor Downregulates MYCN Expression and Promotes Cell Differentiation of Neuroblastoma  [PDF]
Pei-Yi Wu, Yung-Feng Liao, Hsueh-Fen Juan, Hsuan-Cheng Huang, Bo-Jeng Wang, Yen-Lin Lu, I-Shing Yu, Yu-Yin Shih, Yung-Ming Jeng, Wen-Ming Hsu, Hsinyu Lee
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0088795
Abstract: Neuroblastoma (NB) is the most common malignant disease of infancy. MYCN amplification is a prognostic factor for NB and is a sign of highly malignant disease and poor patient prognosis. In this study, we aimed to investigate novel MYCN-related genes and assess how they affect NB cell behavior. The different gene expression found in 10 MYCN amplification NB tumors and 10 tumors with normal MYCN copy number were analyzed using tissue oligonucleotide microarrays. Ingenuity Pathway Analysis was subsequently performed to identify the potential genes involved in MYCN regulation pathways. Aryl hydrocarbon receptor (AHR), a receptor for dioxin-like compounds, was found to be inversely correlated with MYCN expression in NB tissues. This correlation was confirmed in a further 14 human NB samples. Moreover, AHR expression in NB tumors was found to correlate highly with histological grade of differentiation. In vitro studies revealed that AHR overexpression in NB cells induced spontaneous cell differentiation. In addition, it was found that ectopic expression of AHR suppressed MYCN promoter activity resulting in downregulation of MYCN expression. The suppression effect of AHR on the transcription of MYCN was compensated for by E2F1 overexpression, indicating that E2F1 is involved in the AHR-regulating MYCN pathway. Furthermore, AHR shRNA promotes the expression of E2F1 and MYCN in NB cells. These findings suggest that AHR is one of the upstream regulators of MYCN. Through the modulation of E2F1, AHR regulates MYCN gene expression, which may in turn affect NB differentiation.
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