Drug Repurposing of Clinically Approved Drugs to Target Epithelial-mesenchymal Transition Using Molecular Docking Approach

@#Introduction: Epithelial-mesenchymal transition (EMT) is a process of epithelial transformation into mesenchymal cells. It is also a process that contributes to the progression of fibrosis and cancer metastasis. Transforming growth factor-beta (TGF-β), as a potent inducer of EMT, has therefore became a potential therapeutic target. However, clinical developments of TGF-β inhibitors have been un-successful due to safety risks. Hence, drug repurposing of existing safe-to-use drugs could over-come this issue. Methods: In this study, the TGF-β receptor type 1 (ALK5) was selected as the target protein. Molecular docking was performed using known ALK5 inhibitors as positive controls. Clinical drugs with similar binding affinity and amino acid interaction were selected for in vitro experimental validation. Results: ALK5 inhibitor demonstrated binding affinities ranging from -11.2 to -9.5 kcal/mol. Analysis of amino acid interaction revealed that Val219, Ala230, Lys232, and Leu340 amino acid residues are crucial for binding. Subsequent screening of clinically approved drugs against ALK5 showed top five potential drugs (ergotamine, telmisartan, saquinavir, indinavir, and nelfinavir). The selected drugs were tested in TGF-β1-induced normal human bronchial epithelial cell line, BEAS-2B. Western blot analysis showed that the drugs did not exhibit inhibitory effects on the downregulation of epithelial proteins (E-cadherin) and upregulation of mesenchymal proteins (vimentin and α-smooth muscle actin). Conclusion: Based on these experimental outcome, it is postulated that the results from molecular docking were false positives. The tested drugs in this study could serve as negative controls in future screening against ALK5 protein.

Autophagy enhanced by curcumin ameliorates inflammation in atherogenesis via the TFEB-P300-BRD4 axis

Disturbance of macrophage-associated lipid metabolism plays a key role in atherosclerosis. Crosstalk between autophagy deficiency and inflammation response in foam cells (FCs) through epigenetic regulation is still poorly understood. Here, we demonstrate that in macrophages, oxidized low-density lipoprotein (ox-LDL) leads to abnormal crosstalk between autophagy and inflammation, thereby causing aberrant lipid metabolism mediated through a dysfunctional transcription factor EB (TFEB)-P300-bromodomain-containing protein 4 (BRD4) axis. ox-LDL led to macrophage autophagy deficiency along with TFEB cytoplasmic accumulation and increased reactive oxygen species generation. This activated P300 promoted BRD4 binding on the promoter regions of inflammatory genes, consequently contributing to inflammation with atherogenesis. Particularly, ox-LDL activated BRD4-dependent super-enhancer associated with liquid-liquid phase separation (LLPS) on the regulatory regions of inflammatory genes. Curcumin (Cur) prominently restored FCs autophagy by promoting TFEB nuclear translocation, optimizing lipid catabolism, and reducing inflammation. The consequences of P300 and BRD4 on super-enhancer formation and inflammatory response in FCs could be prevented by Cur. Furthermore, the anti-atherogenesis effect of Cur was inhibited by macrophage-specific Brd4 overexpression or Tfeb knock-out in Apoe knock-out mice via bone marrow transplantation. The findings identify a novel TFEB-P300-BRD4 axis and establish a new epigenetic paradigm by which Cur regulates autophagy, inhibits inflammation, and decreases lipid content.

Clinical Effect of Bushen Shengxue Prescription on Chronic Aplastic Anemia and Its Effect on T Cell Subsets and Expression of T-bet and GATA3 / 中国实验方剂学杂志

ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ2=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ2=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ2=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4+， and CD4+/CD8+ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19-， HLA/DR+， and CD25+ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4+， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.

The poly(ADP-ribosyl)ation of BRD4 mediated by PARP1 promoted pathological cardiac hypertrophy

The bromodomain and extraterminal (BET) family member BRD4 is pivotal in the pathogenesis of cardiac hypertrophy. BRD4 induces hypertrophic gene expression by binding to the acetylated chromatin, facilitating the phosphorylation of RNA polymerases II (Pol II) and leading to transcription elongation. The present study identified a novel post-translational modification of BRD4: poly(ADP-ribosyl)ation (PARylation), that was mediated by poly(ADP-ribose)polymerase-1 (PARP1) in cardiac hypertrophy. BRD4 silencing or BET inhibitors JQ1 and MS417 prevented cardiac hypertrophic responses induced by isoproterenol (ISO), whereas overexpression of BRD4 promoted cardiac hypertrophy, confirming the critical role of BRD4 in pathological cardiac hypertrophy. PARP1 was activated in ISO-induced cardiac hypertrophy and facilitated the development of cardiac hypertrophy. BRD4 was involved in the prohypertrophic effect of PARP1, as implied by the observations that BRD4 inhibition or silencing reversed PARP1-induced hypertrophic responses, and that BRD4 overexpression suppressed the anti-hypertrophic effect of PARP1 inhibitors. Interactions of BRD4 and PARP1 were observed by co-immunoprecipitation and immunofluorescence. PARylation of BRD4 induced by PARP1 was investigated by PARylation assays. In response to hypertrophic stimuli like ISO, PARylation level of BRD4 was elevated, along with enhanced interactions between BRD4 and PARP1. By investigating the PARylation of truncation mutants of BRD4, the C-terminal domain (CTD) was identified as the PARylation modification sites of BRD4. PARylation of BRD4 facilitated its binding to the transcription start sites (TSS) of hypertrophic genes, resulting in enhanced phosphorylation of RNA Pol II and transcription activation of hypertrophic genes. The present findings suggest that strategies targeting inhibition of PARP1-BRD4 might have therapeutic potential for pathological cardiac hypertrophy.

Development of hedgehog pathway inhibitors by epigenetically targeting GLI through BET bromodomain for the treatment of medulloblastoma

Medulloblastoma (MB) is a common yet highly heterogeneous childhood malignant brain tumor, however, clinically effective molecular targeted therapy is lacking. Modulation of hedgehog (HH) signaling by epigenetically targeting the transcriptional factors GLI through bromodomain-containing protein 4 (BRD4) has recently spurred new interest as potential treatment of HH-driven MB. Through screening of current clinical BRD4 inhibitors for their inhibitory potency against glioma-associated oncogene homolog (GLI) protein, the BRD4 inhibitor

Design, synthesis, and biological evaluation of quinazolin-4(3H)-one derivatives co-targeting poly(ADP-ribose) polymerase-1 and bromodomain containing protein 4 for breast cancer therapy

@#This study was aimed to design the first dual-target small-molecule inhibitor co-targeting poly (ADP-ribose) polymerase-1 (PARP1) and bromodomain containing protein 4 (BRD4), which had important cross relation in the global network of breast cancer, reflecting the synthetic lethal effect. A series of new BRD4 and PARP1 dual-target inhibitors were discovered and synthesized by fragment-based combinatorial screening and activity assays that together led to the chemical optimization. Among these compounds, 19d was selected and exhibited micromole enzymatic potencies against BRD4 and PARP1, respectively. Compound 19d was further shown to efficiently modulate the expression of BRD4 and PARP1. Subsequently, compound 19d was found to induce breast cancer cell apoptosis and stimulate cell cycle arrest at G1 phase. Following pharmacokinetic studies, compound 19d showed its antitumor activity in breast cancer susceptibility gene 1/2 (BRCA1/2) wild-type MDA-MB-468 and MCF-7 xenograft models without apparent toxicity and loss of body weight. These results together demonstrated that a highly potent dual-targeted inhibitor was successfully synthesized and indicated that co-targeting of BRD4 and PARP1 based on the concept of synthetic lethality would be a promising therapeutic strategy for breast cancer.

Expression of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain on CD8 + T cells in silicosis patients with Mycobacterium tuberculosis infection / 中华传染病杂志

Objective:To explore the expression and clinical significance of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) on the peripheral blood mononuclear cells (PBMC) in silicosis patients with Mycobacterium tuberculosis infection. Methods:August 2018, a total of 78 patients with silicosis (all were quarry workers in Sanmen County, Zhejiang Province) were enrolled and divided into silicosis combined with active pulmonary tuberculosis group (APTB group), silicosis combined with latent tuberculosis infection group (LTBI group), and simple silicosis with non-tuberculosis infection group (non-TB group). Flow cytometry was used to analyze the expressions of TIGIT, programmed death-1 (PD-1) and transcription factor T-bet on PBMC from patients. Mann-Whitney U test and Pearson correlations analysis were used for statistical analysis. Results:Among the 78 patients, eight were in the APTB group, 24 in the LTBI group, and 46 in the non-TB group. The expressions of PD-1 and TIGIT on CD8 + T cells in the APTB group (29.45%(16.78%) and 65.40%(12.12%), respectively) were significantly higher than those in the LTBI group (17.40%(11.17%) and 48.30%(28.75%), respectively; U=23.500 and 43.500, respectively, P=0.000 8 and 0.020 5, respectively) and non-TB group (15.95%(12.46%) and 45.30%(19.75%), respectively; U=64.000 and 69.000, respectively, P=0.002 3 and 0.003 8, respectively), and the differences were all statistically significant. The expression of TIGIT was positively correlated with PD-1 on CD8 + T cells in silicosis patients ( r=0.434 3, P<0.01). The proportion of PD-1 + TIGIT + CD8 + T cells in the APTB group (19.90%(22.67%)) was significantly higher than those in the non-TB group (11.55%(11.29%), U=76.500, P=0.007 1) and LTBI group (11.55%(10.53%), U=41.000, P=0.015 4), while the proportion of PD-1 -TIGIT -CD8 + T cells in the APTB group (30.60%(12.90%)) was significantly lower than non-TB group (48.90%(18.98%), U=58.000, P=0.001 3) and LTBI group (47.20%(24.59%), U=41.000, P=0.015 4). The differences were all statistically significant. The expression of T-bet on the peripheral blood CD8 + T cells in the APTB group (29.45%(16.78%)) was higher than that in the non-TB group (15.95%(12.46%)) and the LTBI group (17.40%(11.17%)), and the differences were both statistically significant ( U=46.500 and 46.000, respectively, P=0.000 3 and 0.028 3, respectively). The expression of T-bet on CD8 + T cells was positively correlated with TIGIT on CD8 + T cells ( r=0.456 7, P<0.01). The expression of T-bet on PD-1 + TIGIT + CD8 + T cells in the APTB group (65.40%(12.12%)) was higher than those in the LTBI group (48.30%(28.75%), U=23.500, P=0.000 8) and non-TB group (45.30%(19.75%), U=65.000, P=0.002 6), and the differences were both statistically significant. Conclusion:The immunosuppressive receptor PD-1 and TIGIT are highly expressed on CD8 + T cells in silicosis patients with active pulmonary tuberculosis, which indicates CD8 + T cells exhaustion in these population, while the highly co-expression of T-bet suggests the exhausted subsets may have reversed potentiality.

A homogenous nanoporous pulmonary drug delivery system based on metal-organic frameworks with fine aerosolization performance and good compatibility

Pulmonary drug delivery has attracted increasing attention in biomedicine, and porous particles can effectively enhance the aerosolization performance and bioavailability of drugs. However, the existing methods for preparing porous particles using porogens have several drawbacks, such as the inhomogeneous and uncontrollable pores, drug leakage, and high risk of fragmentation. In this study, a series of cyclodextrin-based metal-organic framework (CD-MOF) particles containing homogenous nanopores were delicately engineered without porogens. Compared with commercial inhalation carrier, CD-MOF showed excellent aerosolization performance because of the homogenous nanoporous structure. The great biocompatibility of CD-MOF in pulmonary delivery was also confirmed by a series of experiments, including cytotoxicity assay, hemolysis ratio test, lung function evaluation,

Production of activated carbon from mango seed coat using chemical activation

Aim: This study was carried out find the feasibility of utilizing raw mango seed coat under various process conditions to produce the best activated carbon by chemical activation and compared with commercial activated carbon. Methodology: Activated carbon was produced by chemical activation under various process conditions such as different activation temperatures, activating agents, impregnation volume percentages and activation times for pyrolysis in a programmable electrical furnace with reactor in the absence of air. Results: The results were compared using phosphoric acid having 50% impregnation volume to other activating agents, the activating temperature was 400oC, activation time 1 hr, iodine number, methylene blue number, % yield and B.E.T surface area being 831 mg g-1, 212 mg g-1, 41.09% and 1114 m2 g-1 respectively. Interpretation: Carbon samples prepared using mango seed coat treated by H3PO4, showed clear open porous structures along with a larger pore size compared to commercially available activated carbon.

Effect of Modified Erchentang on GATA3 and T-bet mRNA Expressions in Lung Tissue of Chronic Obstructive Pulmonary Disease Rats / 中国实验方剂学杂志

Objective: To observe the effect of modified Erchentang on GATA-binding protein-3(GATA3) and T-box expressed in T cells(T-bet) in lung tissue of rats with chronic obstructive pulmonary disease (COPD). Method: Seventy SD rats were randomly divided into seven groups, namely normal group, model group, low, medium and high-dose modified Erchentang group(5,10,20 g ·kg-1), Xiaokechuan group(5 g ·kg-1) and Erchentang group(5 g ·kg-1), with 10 in each group. The rat model of COPD was established by smoking combined with intratracheal dripping of lipopolysaccharide (LPS). After successful modeling, the treatment group was given intragastric administration, and the normal group and the model group were given intragastric administration of equal volume of saline. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentrations of interleukin-10 (IL-10) and interleukin-12 (IL-12) in rat serum. The expressions of GATA3 and T-bet were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The expressions of GATA3 and T-bet in lung tissue were detected by immunohistochemistry (IHC). Result: Compared with the control group, the serum levels of IL-10 in the model group was significantly decreased, while the IL-12 level was significantly increased (PPPPConclusion: Modified Erchentang may reduce the inflammation of lung tissue and improve lung function in COPD rats by reducing IL-12, increasing the content of IL-10, inhibiting the protein and gene expressions of T-bet, and stimulating the protein and gene expressions of GATA3.

Oral Allergy Syndrome in Birch Pollen-Sensitized Patients from a Korean University Hospital

BACKGROUND: Oral allergy syndrome (OAS) is a type of allergic reaction that mainly occurs on oral contact with raw fruit, vegetables, or nuts. The most common type of OAS is birch pollen-related food allergy. Although OAS is a common food allergy in adults, only few epidemiologic studies have been reported in Korea. Here we investigate the prevalence and triggers of birch pollen-related food allergy. METHODS: We conducted a retrospective chart review of 1,427 patients who underwent a skin prick test for inhalant allergens at the Asthma and Allergy Clinic in Seoul National University Bundang Hospital from January 2011 to December 2016. RESULTS: Of 1,427 patients, 125 (8.7%) were sensitized to birch pollen. Among them, 20.0% developed OAS, which was the most common food allergy (96.2%). The prevalence of OAS was higher in females, and was 18.2% in birch pollen-sensitized allergic rhinoconjunctivitis patients. Further, 72.0% OAS patients had rhinoconjunctivitis, 20.0% had asthma, and 12.0% had chronic urticaria. Apple (68.0%), peach (56.0%), nuts (36.0%), kiwi (20.0%), persimmon (20.0%), plum (16.0%), and cherry (16.0%) were frequent triggers; however, Chinese yam, kudzu vine, bellflower root, codonopsis, and ginseng were also revealed as triggers. Patients (60.0%) showed OAS with ≥ 3 foods at the same time. Only 3 patients showed mono-sensitivity to birch pollen, while others were multi-sensitized to trees, grasses, weed, or house dust mite allergens. CONCLUSION: OAS was the most common food allergy in birch pollen-sensitized patients. This study revealed the unique triggers of OAS in Korea in addition to well-known triggers.

Study on the expression and significance of Th1/Th2 cells in the blood of patients with primary gouty arthritis / 中华风湿病学杂志

Objective This study is aimed to investigate the possible role of Th1/Th2 cell in the pathogenesis of primary gout arthritis. Methods The peripheral blood of 21 acute gout patients (AG), 20 intermittent gout patients (IG) and 20 healthy controls (HC) were collected. The clinical data and laboratory indicators of them were enrolled. The percentages of Th1 and Th2 cells were detected by flow cytometry (FCM). The expression of GATA-3, T-bet, IL-4 and interferon (IFN)-γ mRNA in Peripheral blood mononuclear cells (PBMCs) were measured using Real time quantitative polymerase chain reaction (PCR). The protein expression levels of IL-4 and IFN-γ in serum were detected by enzyme-linked immuno sorbent assay (ELISA). The measurement data were compared by one factor analysis of variance test. The correlation between variables was used by Spearman correlation analysis. Results The percentage of Th1 cells in peripheral blood of the AG group was [(23.2 ±8.3)％], and the IG group was [(20.5 ±9.3)％], which were significantly higher (F=6.520, P<0.05) than the percentage of HC group [(14.8±3.8)％]. There was no significant difference between AG group and IG group. The percentage of Th2 cells in peripheral blood of AG group were [(1.9 ±0.7)％], which was significantly lower (F=8.267, P<0.05) than and the percentage of HC group [(3.4±1.8)％] and IG group [(3.3± 1.2)％]. There was no significant difference between the IG group and the HC group. And the proportion of TH1/Th2 cells in the AG group was higher than that of the IG group and the HC group (F=10.406, P<0.01). The expression of T-bet mRNA and IFN-γ mRNA in the AG group and IG group were higher than that in the HC group (F=4.942, P=0.010)、(F=4.458, P=0.016). However, there was no significant difference between IG group and AG group. The expression of GATA-3 mRNA and IL-4 mRNA was significantly lower (F=3.564, P=0.035) (F=5.385, P=0.007) in the AG group and IG group when compared to the HC group. And there was no significant difference between the AG and the IG group. The IFN-γ level increased in the AG and IG group compared to the HC group (F=7.659, P=0.001). The IL-4 levels in the AG group was lower (F=7.099, P=0.002) than those of the IG and HC group. Conclusion Th1 cells in the peripheral blood of patients with GA are increased and accompanied with the decrease of Th2 cells. The results of this study suggest that the imbalance of Th1/Th2 cells in the inflammatory and immune response plays a critical role in the pathogenesis of GA.

Influence of T-bet/GATA-3 to Th1/Th2 of athma rats / 中国免疫学杂志

Objective:To discuss the influence of dexamethasone(DXM) to T-bet/GATA-3 of athma rats.Methods: 30 SD rats were randomly divided into normal group,model group and DXM group,10 rats in each group.Ovalbumin was intraperitoneal injected on the 1th and 8th day and aerosol inhaled from the 15th day,once a day,14 days altogether.Dexamethasone was intraperitoneal injected in 0.5 mg/kg from the 15th day,once per day,a total of 14 times.Flow cytometry tested the content of Th1 and Th2 in spleen and peripheral blood,immunohistochemistry and Western blot measured T-bet and GATA-3 protein expression and RT-PCR tested T-bet and GATA-3 mRNA expression.Results: Compared model group with normal group,the contents of Th1,Th2 in spleen and peripheral blood had a very significant increase(P<0.01);expression of T-bet,GATA-3 protein and mRNA in the lung tissue also had very significant risen(P<0.01).While compared DXM group with model group,T-bet/GATA-3 rose significantly(P<0.05)or very significantly(P<0.01).Conclusion: T-bet/GATA-3 is the key transcription factors that influence the balance of Th1/Th2,while DXM can raise the ratio of T-bet/GATA-3,which reduce the severity of asthma.

BET Bromodomain inhibitors and degraders based on polypharmacology:research advances / 国际药学研究杂志

Bromodomain and extra-terminal domain(BET)Bromodomain has become a new target for the treatment of cancers and other human disorders. Nowadays,several classes of its potent and selective small-molecule inhibitors have been identified,many of which are in clinical trials. Preclinical and clinical data have shown that BET Bromodomain inhibitors have good prospects. Howev-er,there are potential therapeutic deficiencies,such as drug resistance. At present,attempts are being made to develop BET Bromodo-main inhibitors and degraders based on polypharmacology,combining BET Bromodomain with other targets of different mechanisms. In this paper,small-molecule kinase/BET inhibitors,small-molecule histone deacetylases(HDAC)/BET inhibitors and BET protein degraders are reviewed,which may provide guidance for further research on BET protein.

BET Bromodomain Involves in Inflammatory Genes Transcription via Regulation of NF-κB Signaling Pathway / 现代生物医学进展

Objective:To explore bromodomain and extra-terminal (BET) inhibition in the regulation of vascular endothelial cells activation and early atherosclerosis formation and its potential molecular mechanisms.Methods:1.Human umbilical vein endothelial cells (HUVEC) and mouse heart endothelial cells (MHEC) were isolated,and tumor necrosis factor α (TNFα) was used to activate in flammatory genes transcription in the presence or absence of JQ1,a specific BET inhibitor.The groups are as follows:(1)Normal control group;(2) TNFα(25 ng/mL)group;(3) TNFα+JQ 1 group.The gene mRNA and protein expression of inflammatory cytokines were measured by both real-time PCR and flow cytometry (FCM).2.LDL receptor-deficient (LDLR-/-) mice were randomly divided into 2 groups:JQ1 group (n=8,JQlintraperitoneal,50 mg/kg,daily) and control group (n=8,DMSO,daily).After 8 weeks feeding with high cholesterol diet,vascular cell adhesion molecule-1 (VCAM-1) expression in aortic arch was measured by immunohistochemistry.The activity of nuclear factor kappa B (NF-κB) signaling was monitored by 5XκB luciferase reporter assay in HEK293.Results:TNFα dramatically induced the mRNA and protein expression of inflammatory genes and JQ 1 significantly downregulated the induction of them (E-selectin,P-selectin,VCAM-1,IL-8)(P＜0.01).Immunohistochemistry detection indicated that JQ1 significantly downregulated the expression of VCAM-1 in aortic arch induced by 8 weeks high cholesterol diet feeding comparing to control group.In addition,BET bromodomain inhibition downregulated TNFα upregulated NF-κB transcriptional activity (P＜0.01).Conclusions:Our study demonstrated that BET bromodomain was involved in NF-κB mediated inflammatory genes expression;inhibition of BET bromodomain suppressed vascular endothelial activation in vitro,and attenuated early atherogenesis in vivo.

Intervenient study of Shengjiang San on Th1/Th2 imbalances and related regulatory factors in sepsis pa- tients / 实用医学杂志

Objective To investigate the intervention of Chinese herb Shengjiang San(SJS)for the imbal-ances of serum Th1/Th2 cells and related regulatory factors in sepsis patients. Methods Fifty-five sepsis patients were randomly divided into two groups:conventional treatment group of 27 cases and SJS group of 28 cases. Nine cases of healthy volunteers were enrolled as control group in the study. Cases of conventional group were treated with western medicine only and cases of SJS group were treated with both western medicine and Chinese herb SJS (100 mL twice one day). The therapy course of both groups was 3 days. Score of Chinese medical syndromes ,se-rum level of leukocyte count,C reactive protein(CRP)and procalcitonin(PCT),serum T-bet,GATA-3,Th1&Th2 cells in the proportion of whole CD4+Th cells and Th1/Th2 ratio were compared respectively in each group be-fore and after treatment. Results There were significant differences between SJS group and conventional group in score of Chinese medical syndromes,serum level of leukocyte count,T-bet,GATA-3,Th1&Th2 cells in the pro-portion of whole CD4+Th cells and Th1/Th2 ratio(P<0.05 or P<0.01). There were no significant differences be-tween the two groups in serum level of CRP,PCT and GATA3. Compared with control group,there were signifi-cant differences in all indicators except GATA3 of the two groups(P<0.01). Conclusion Chinese herb Shengji-ang San has an effective benefits to Chinese medical syndromes,inflammatory reaction,the imbalance of serum Th1/T2 and related regulatory factors(T-bet&GATA-3)in sepsis patients.

Expression and meaning of T-bet and GATA3 mRNA in T lymphocyte of patients during the operation with cardiopulmonary bypass / 中华危重病急救医学

Objective To investigate the effects of cardiopulmonary bypass (CPB) on the differentiation of T lymphocyte subsets and the expression of specific transcription regulator T-bet/GATA binding protein 3 (GATA3). Methods A prospective double-blind study was conducted. Patients with CPB pulmonary repair of ventricular septal defect (observation group) or off-pump ligation of ductus arteriosus (control group) with 20 cases each in the 150th Military Hospital from February 2015 to February 2016 were enrolled. The blood sampled was collected on the time of before operation, at the end of CPB or operation, 4 hours after operation, and 24 hours after operation. T lymphocytes were isolated, the helper T cell 1 (Th1) specific transcription factor T-bet mRNA, helper T cell 2 (Th2) specific transcription factor GATA3 mRNA expression and cytokine γ-interferon (IFN-γ) mRNA, interleukin-4 (IL-4) mRNA expression were measured by Northern Blot. Results Compared with before operation, expression levels of T-bet mRNA [integral gray values: (1.39±0.52)×105vs. (2.92±0.88)×105], IFN-γ mRNA [integral gray values: (3.68±0.65)×105vs. (6.10±0.93)×105] were decreased transiently at the end of CPB in the observation group (both P < 0.05), returned to preoperative levels at 24 hours after operation [integral gray values: (2.77±0.74)×105, (6.22±1.25)×105, respectively, both P > 0.05]; expression levels of GATA3 mRNA [integral gray values:(4.96±0.88)×105vs. (3.21±0.68)×105], IL-4 mRNA [integral gray values: (3.52±1.13)×105vs. (1.85±0.63)×105] were increased (both P < 0.05), recovered to the preoperative levels at 24 hours after operation [integral gray values: (3.11±0.51)×105, (1.93±0.84)×105, respectively, both P > 0.05]. There were no significant differences in the expressions of T-bet, GATA3, IFN-γ and IL-4 mRNA in the control group at each time points (all P >0.05). Conclusions CPB causes the imbalance of Th1, Tc1/Th2, Tc2 and pro-inflammatory and anti-inflammatory reactions specially, which participate the complication occurrence after CPB. The changing of T-bet/GATA3 may be the internal mechanism for these changes.

Effects of arsenic trioxide on T-bet/GATA3 signal pathway in MRL/lpr mice / 中国病理生理杂志

AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.

Quercetin improves Pseudomonas aeruginosa-induced lung infection in rats / 中成药

AIM To study the improvement of quercetin on Pseudomonas aeruginosa-induced lung infection in rats.METHODS Forty SPF SD rats were randomly divided into five groups,eight rats in each group:normal group,P.aeruginosa infection group,quercetin group,levofloxacin group,levofloxacin combined with quercetin group (combined group),the rats were anesthetized and then injected with P.aeruginosa in bronchus.The pathological changes of lung tissue in rats were observed,the contents of IL-4 and IFN-γcytokines,changes of transcription factors T-bet and Gata-3 in lung tissue were detected,and then semi-quantitative RT-PCR was used for the detection of IL-4,IFN-γ,T-bet and Gata-3 mRNA expressions.RESULTS The content of IL-4,levels of IL-4 and Gata-3 mRNA in lung tissue in the quercetin group,the levofloxacin group and the combined group were lower than those in the P.aeruginosa infection group,but the opposite was true in the levels of IFN-γ and T-bet mRNA.CONCLUSION Quercetin and levofloxacin can induce the differentiation from type Th2 to type Th1 for rat organism,but there is no synergistic effect between them.

Effects of arsenic trioxide on T-bet/GATA3 signal pathway in MRL/lpr mice / 中国病理生理杂志

AIM:To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS:MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups,respectively.The mice in 2 sub-groups received ATO (0.4 mg · kg-1.d-1) and sodium chloride (NS,volume weight-determined) by intraperitoneal injection respectively for 2 months.Afterward,the spleens were isolated fron the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared.The mRNA level of T-bet,GATA3,IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR.The protein expression of T-bet and GATA3 was determined by Western blot.The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS:The mRNA and protein levels of T-bet,IFN-γand the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P ＜0.05).However,the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P ＜ 0.05).In MRL/lpr mice,the mRNA and protein levels of T-bet,IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P ＜ 0.05),no difference was found in GATA3 and IL-4.No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION:ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.