Tiaobu Feishen therapy inhibits inflammation induced by cigarette smoke extracts in a human monocyte/macrophage cell line.

OBJECTIVE: To study the mechanistic effects of Tiaobu Feishen therapy (TBFS) on inflammation induced by cigarette smoke extract (CSE) in a human monocyte/macrophage cell line. METHODS: The human monocyte/macrophage cell line THP-1 was stimulated with 10 % CSE in the presence or absence of Bufei Yishen formula (BYF), Bufei Jianpi formula (BJF) and Yiqi Zishen formula (YZF). All formulations contained serum. Pro-inflammatory cytokines were measured in the supernatants using enzyme-linked immunosorbent assay. The activity of STAT3 DNA binding was detected using electrophoretic mobility shift assay and janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway activation was assessed using Western blotting. RESULTS: The results showed that BYF, BJF and YZF treatment strongly decreased the CSE-induced secretion of interleukin (IL)-6, IL-8, tumor necrosis factor-α and matrix metalloproteinase-9 by THP-1 cells. Furthermore, BYF, BJF and YZF treatment attenuated STAT3 DNA binding capacity and JAK2 and STAT3 were shown to be phosphorylated. CONCLUSION: The data revealed that BYF, BJF and YZF effectively inhibited a CSE-induced inflammatory response in THP-1 cells by limiting activation of the JAK2/STAT3 pathway.

N-acetylcysteine decreases malignant characteristics of glioblastoma cells by inhibiting Notch2 signaling.

BACKGROUND: Glioblastomas multiforme (GBM) is the most devastating primary intracranial malignancy lacking effective clinical treatments. Notch2 has been established to be a prognostic marker and probably involved in GBM malignant progression. N-acetylcysteine (NAC), a precursor of intracellular glutathione (GSH), has been widely implicated in prevention and therapy of several cancers. However, the role of NAC in GBM remains unclear and the property of NAC independent of its antioxidation is largely unknown. METHODS: The mRNA and protein levels of Notch family and other related factors were detected by RT-PCR and western blot, respectively. In addition, intracellular reactive oxygen species (ROS) was measured by flow cytometry-based DCFH-DA. Moreover, cell viability was assessed by CCK8 and cell cycle was analyzed by flow cytometry-based PI staining. The level of apoptosis was checked by flow cytometry-based Annexin V/PI. Cell migration and invasion were evaluated by wound healing and transwell invasion assays. At last, U87 Xenograft model was established to confirm whether NAC could restrain the growth of tumor. RESULTS: Our data showed that NAC could decrease the protein level of Notch2. Meanwhile, NAC had a decreasing effect on the mRNA and protein levels of its downstream targets Hes1 and Hey1. These effects caused by NAC were independent of cellular GSH and ROS levels. The mechanism of NAC-mediated Notch2 reduction was elucidated by promoting Notch2 degradation through Itch-dependent lysosome pathway. Furthermore, NAC could prevent proliferation, migration, and invasion and might induce apoptosis in GBM cells via targeting Notch2. Significantly, NAC could suppress the growth of tumor in vivo. CONCLUSIONS: NAC could facilitate Notch2 degradation through lysosomal pathway in an antioxidant-independent manner, thus attenuating Notch2 malignant signaling in GBM cells. The remarkable ability of NAC to inhibit cancer cell proliferation and tumor growth may implicate a novel application of NAC on GBM therapy.

[Effect of Qige Powder on Angiogenesis Induced by Esophageal Cancer Cell Line Eca-9706].

OBJECTIVE: To study the effect Qige powder on esophageal cancer angiogenesis. METHODS: Inhibitive effect of Qige powder ethanol extract on proliferation of Esophageal cancer cell line Eca-9706 was detected by MTT assay. The Eca-9706 cells conditioned medium in Qige powder IC50 concentration were collected. Angiogenesis, as well as proliferation, migration and tube formation of human umbilical vein endothelial cells were observed by Chick embryo chorioallantoic membrane model(CAM), MTT assay, the migration and tube formation assay. VEGF and IL-6 contents in culture medium supernatant of Eca-9706 cells and human umbilical vein endothelial cells were detected by ELISA. RESULTS: Qige powder ethanol extract could inhibit the proliferation of Eca-9706 cells, with a certain dose-effect relationship, IC50 value was 96 µg/mL. Eca-9706 cells conditioned medium could significantly increase the CAM generating total vessel area, human umbilical vein endothelial cells proliferation,migration and tube formation, while the Eca-9706 cell conditioned medium of Qige powder ethanol extract could reduce CAM angiogenesis, human umbilical vein endothelial cells proliferation and tube formation, but increase endothelial cells migration. Qige powder ethanol extract could reduce endothelial cells secreting VEGF and IL-6 while increase Eca-9706 cells secreting. CONCLUSION: Qige powder ethanol extract can inhibit angiogenesis, endothelial cell proliferation, and tube formation induced by Eca-9706 cells, while reduce VEGF and IL-6 secretion of endothelial cells.

[Discussion of Chinese medical research ideas from miRNAs features].

In this paper, miRNAs features were briefly introduced and agreeable points were discussed from 4 aspects: organs relationship, syndrome research, Chinese medical pathogeneses, and actions of Chinese herbs. miRNAs, as information media for organs interrelation, was believed to explain Chinese medical pathogeneses and reveal partial molecular mechanisms of Chinese medicine. miRNAs in the body fluid could be taken as one of biological bases of syndromes.

[The relationship between inhibitory effect of xuanfuguilian prescription on the growth of esophageal cells ECA9706 and its components].

OBJECTIVE: To discuss the relationship between inhibitory effect of different concentration ethanol extracts from Xuanfuguilian prescription on the growth of esophageal cells ECA9706 and its components. METHODS: The drug was extracted with anhydrous ethanol, 95%, 85%, 75%, 50%, 25% ethanol or water, the cell proliferation inhibitory rate of different solvent extracts were detected with MTT assay, and IC50 was calculated. The components of the drug were determined by LC/MS. Based on the inhibitory rate and the components peak area, the samples were hierarchy clustered respectively. The correlation of components peak area and inhibition rate was analyzed with Pearson Correlation. The components peak area and inhibition rate were analyzed using PLS. RESULTS: The IC50 of the 7 extracts on esophageal carcinoma cell ECA9706 were respectively 46.361, 52.67, 58.11, 78.26, 93.10, 2579.43 and 3953.34 microg/ mL 22 stable peaks were determined by LC-MS. Based on the inhibition rate and the components peak area, the clustering results of the two samples were similar. The 10 peaks areaes were closely related to inhibition rate (P < 0.05) and the PLS between components peaks area and inhibition rate was constructed. CONCLUSION: Extracts with different concentration ethanol have different effects, and their curative effects are closely related to components.

[Effect of invigorating lung and kidney prescription and its components on secretion of cytokines induced by CSE and LPS].

OBJECTIVE: To observe effect of Invigorating Lung and Kidney Prescription (ILKP) on secretion of cytokines induced by Lipopolysaccharide (LPS) and cigarette smoke extract (CSE) in vitro and discuss prescription regularity of ILKP and reveal molecular mechanism of ILKP treatment on COPD. METHODS: Alveolar epithelial cells A549, airway epithelial cells H292 and monocyte macrophage THP-1 were stimulated with LPS and CSE, in which samples were divided into normal group (20% normal rat serum), model group (20% normal rat serum and 5% CSE with 75 pg/mL LPS), ILKP group and its Components [20% Replenishing qi group (1), Replenishing Lung and Kidney group (2), Activating blood group (3), Reduce phlegm group (4), Replenishing qi with Replenishing Lung and Kidney group (5), Replenishing qi with Activating blood group (6), Replenishing qi with Reduce phlegm group (7), Replenishing qi with Replenishing Lung and Kidney and Activating blood group (8), Replenishing qi with Replenishing Lung and Kidney and Reduce phlegm group (9), Replenishing qi with Activating blood and Reduce phlegm group (10), ILKP serum group (11)]. The cytokines were detected by ELISA and the transcription factors of cytokines were analyzed with Network tool. RESULTS: Compared with the normal group,the secretion of MMP-9, GM-CSF and TGF-beta by THP-1 cells,TIMP by A549 cells,TNF-alpha, MMP-9, GM-CSF, CD36 and TIMP-1 by H292 cells all were increased, while secretion of TNF-alpha, PPAP2B, IL-3 and SOD decreased by THP-1 cells, MMP-9, TNF-alpha, TGF-beta and IL-3 by A549 cells, IL-8 by H292 cells all were decreased. Compared with model group, for THP-1 cells, the secretion of GM-CSF in 3, 6, 10 and 11 prescription groups,TGF-beta in 10 and 11 prescription groups all were decreased. MMP-9 decreased in 9 and 5 prescription groups while it increased in 1, 2, 3, 4, 7, 8, 10 and 11 prescription groups. TNF-alpha was increased in 1, 2, 3, 4, 6, 8, and 9 prescription groups, as it decreased in 11 prescription group. Except 2 prescription group, IL-3 was increased in all other drugs groups. SOD in 2, 4, 5, 8, 9, 11 prescription groups, and PPAP2B in 1, 2, 3, 4, 5, 6, 7 prescription groups were increased while PPAP2B was decreased in 11 prescription group. For A549 cells, the secretion of TIMP-1 was increased in 1, 2, 3, 4, and 5 prescription groups, while it was decreased in 11 prescription groups; Except 1 and 4 prescription group, MMP-9 was increased in other groups; Except TGF-beta was increased in 11 prescription groups, it was increased in other groups; IL-3 was increased in 1, 2, 4, 5, 11 and 3 prescription groups. For H292 cells, the secretion of GM-CSF in 5,6,7 and 2 prescription groups,TNF-alpha in 1, 2, 4, 6, 7, 8, 9 and 10 prescription groups, MMP-9 in 4, 8, 7 and 9 prescription groups all were decreased. CONCLUSION: The effect of ILKP and its components on secretion of cytokines induced by LPS and CSE and its target cells are different, in which Activating blood prescription focuses on inflammation cytokines, Replenishing Lung and Kidney prescription on Lipid metabolism and redox regulation. The effect of ILKPA is the most widely, followed by Activating blood Replenishing Lung and Kidney, and Reduce phlegm prescription on Protease regulation.