Effect of koumine on the inflammation of RAW264.7 cells induced by LPS / 中国兽医学报

To reveal the anti-inflammation mechanism of koumine,we determined effects of different concentrations of koumine(100,200,400 mg/L) on the secretion of NO,IL-1β,IL-6 and TNF-a by nitrate reductase and ELISA as well as the mRNA of iNOS,IL-1β,IL-6 and TNF-α.Western blot was used to detect iNOS protein expression.The results showed that 100,200,400 mg/L of koumine can significantly inhibit the secretion of NO,IL-1β,IL-6 and TNF-α of RAW264.7 cell (P＜0.01).Koumine can dose-dependently down-regulate the mRNA of iNOS,IL-1β,IL-6 and TNF-α,meanwhile koumine can also significantly inhibit the protein expression of iNOS.The results indicated that koumine may play an anti-inflammation activity by mean of the reduction of NO and mediator of inflammation,down-regulating the mRNA expression of iNOS,IL-1β,IL-6 and TNF-a,decreasing protein expressions of iNOS.

Effects of Jiedu Quyu Ziyin Decoction on TLR4 Signal Pathway in Lung, Spleen and Peritoneal Macrophagocyte of MRL/lpr Lupus Mice Treated by Prednisone / 浙江中医药大学学报

[Objective] To observe the effect of Jiedu Quyu Ziyin decoction on TLR4 signaling pathway in macrophages of MRL/lpr lupus mice. [Method] MRL/lpr lupus mice were divided into four groups:model group, prednisone group, Jiedu Quyu Ziyin decoction group(hereinafter referred to as:Chinese medicine group) and prednisone plus Chinese medicine group (hereinafter referred to as:combination of Chinese and western medicine group). The mice were gavaged with saline, prednisone, Jiedu Quyu Ziyin decoction and prednisone added Jiedu Quyu Ziyin decoction for 4 weeks. Macrophages of lung, peritoneal and spleen were collected and the expression of related genes was detected by RT-PCR. [Result] TLR4 mRNA in lung macrophages, and TLR4 protein in splenic macrophages increased significantly(P<0.05) after the treatment of prednisone. The increased TLR4 protein in splenic macrophages was significantly decreased by combining with Jiedu Quyu Ziyin decoction(P<0.05). Prednisone can significantly reduce the TLR4 downstream molecules such as MyD88, IFN-α, iNOS mRNA expression in lung and peritoneal macrophages(P<0.05). The decreased MyD88 mRNA in lung macrophages was increased significantly by combining with Jiedu Quyu Ziyin decoction(P<0.05). [Conclusion] TLR4 signaling pathway is changed in macrophages after glucocorticoid administration in MRL/lpr lupus mice. Jiedu Quyu Ziyin decoction can reduce the abnormal glucocorticoid-induced TLR4 protein expression.

Crystalline Silica Promotes Rat Fibrocyte Differentiation in Vitro, and Fibrocytes Participate in Silicosis in Vivo / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>The aim of this study was to investigate the effects of SiO2 on fibrocytes and whether fibrocytes participate in silicosis in vivo.</p><p><b>METHODS</b>A macrophagocyte (AM)/fibrocyte coculture system was established, and AMs were treated with 100 μg/mL SiO2. Flow cytometry was used to detect the number of fibrocytes. Real-time PCR was performed to measure the expression of collagen I, collagen III, and α-SMA mRNA. The levels of collagen I, collagen III, and TGF-β1 protein were determined by ELISA. Immunohistochemical staining was performed to measure α-SMA protein expression. A rat silicosis model was induced by intratracheal instillation of SiO2. Lung histopathological evaluation was conducted using HE and Masson's trichrome staining after 1 and 9 weeks. The number of fibrocytes in peripheral blood or lung tissue of rat was detected by flow cytometry. Double-color immunofluorescence was applied to identify fibrocytes in the lung tissue.</p><p><b>RESULTS</b>Peripheral blood monocytes were found to differentiate into fibrocytes in vitro in a time-dependent manner, and exposure to crystalline silica might potentiate fibrocyte differentiation. In addition, fibrocytes were able to migrate from peripheral blood to the lung tissue, and the number of fibrocytes was increased after SiO2 exposure.</p><p><b>CONCLUSION</b>Silica exposure potentiates fibrocyte differentiation, and fibrocytes may participate in silicosis in vivo.</p>

Expression level of miR-155 in peripheral blood

OBJECTIVE@#To investigate the relationship between the expression level of miR-155 and the severity of coronary lesion, and explore the action mechanism.@*METHODS@#Peripheral blood mononuclear cells (PBMC) were isolated form blood simple from patients with acute myocardial infarction (AMI), unstable angina (UAP), stable angina (SAP) and chest pain syndrome (CPS). RT-PCR was performed to analysis the expression level of miR-155 in peripheral blood mononuclear cells, plasma and RAW264.7 macrophagocyte. MTT was used to analyze the cell viability of OxLDL treated RAW264.7 macrophagocyte.@*RESULTS@#The expression level of miR-155 in blood sample from coronary heart disease patients was much lower than in the blood sample of non-coronary heart disease (P<0.05). The level of miR-155 in PBMCs was much higher in the blood sample from CPS group than the other three group, and the level of miR-155 in plasma was higher in the CPS group than in the UAP and the AMI group, the difference was statistically significant (P<0.05). The expression level of miR-155 in PBMCs is positively associated with the level in the plasma (r=0.861, P=0.000). OxLDL can induce the expression of miR-155 in RAW264.7 macrophagocyte, decrease the cell viability of RAW264.7 macrophagocyte, and with the concentration and the treatment time of OxLDL increased, the effort become more obvious. The inhibition effort of OxLDL to RAW264.7 macrophagocyte with high miR-155 expression is much lower than the control group, and it is statistically significant after treated for 12, 24 and 48 h.@*CONCLUSIONS@#miR-155 plays a protective role in the progression of atherosclerosis, and it may be achieved by reducing the apoptosis effort of OxLDL to RAW264.7 macrophagocyte.

Expression level of miR-155 in peripheral blood

Objective:To investigate the relationship between the expression level of miR-155 and the severity of coronary lesion, and explore the action mechanism.Methods: Peripheral blood mononuclear cells (PBMC) were isolated form blood simple from patients with acute myocardial infarction (AMI), unstable angina (UAP), stable angina (SAP) and chest pain syndrome (CPS). RT-PCR was performed to analysis the expression level of miR-155 in peripheral blood mononuclear cells, plasma and RAW264.7 macrophagocyte. MTT was used to analyze the cell viability of OxLDL treated RAW264.7 macrophagocyte.Results: The expression level of miR-155 in blood sample from coronary heart disease patients was much lower than in the blood sample of non-coronary heart disease (P<0.05). The level of miR-155 in PBMCs was much higher in the blood sample from CPS group than the other three group, and the level of miR-155 in plasma was higher in the CPS group than in the UAP and the AMI group, the difference was statistically significant (P<0.05). The expression level of miR-155 in PBMCs is positively associated with the level in the plasma (r=0.861,P=0.000). OxLDL can induce the expression of miR-155 in RAW264.7 macrophagocyte, decrease the cell viability of RAW264.7 macrophagocyte, and with the concentration and the treatment time of OxLDL increased, the effort become more obvious. The inhibition effort of OxLDL to RAW264.7 macrophagocyte with high miR-155 expression is much lower than the control group, and it is statistically significant after treated for 12, 24 and 48 h.Conclusions: miR-155 plays a protective role in the progression of atherosclerosis, and it may be achieved by reducing the apoptosis effort of OxLDL to RAW264.7 macrophagocyte.

Expression level of miR-155 in peripheral blood

Objective: To investigate the relationship between the expression level of miR-155 and the severity of coronary lesion, and explore the action mechanism. Methods: Peripheral blood mononuclear cells (PBMC) were isolated form blood simple from patients with acute myocardial infarction (AMI), unstable angina (UAP), stable angina (SAP) and chest pain syndrome (CPS). RT-PCR was performed to analysis the expression level of miR-155 in peripheral blood mononuclear cells, plasma and RAW264.7 macrophagocyte. MTT was used to analyze the cell viability of OxLDL treated RAW264.7 macrophagocyte. Results: The expression level of miR-155 in blood sample from coronary heart disease patients was much lower than in the blood sample of non-coronary heart disease (. P<0.05). The level of miR-155 in PBMCs was much higher in the blood sample from CPS group than the other three group, and the level of miR-155 in plasma was higher in the CPS group than in the UAP and the AMI group, the difference was statistically significant (. P<0.05). The expression level of miR-155 in PBMCs is positively associated with the level in the plasma (. r=0.861, P=0.000). OxLDL can induce the expression of miR-155 in RAW264.7 macrophagocyte, decrease the cell viability of RAW264.7 macrophagocyte, and with the concentration and the treatment time of OxLDL increased, the effort become more obvious. The inhibition effort of OxLDL to RAW264.7 macrophagocyte with high miR-155 expression is much lower than the control group, and it is statistically significant after treated for 12, 24 and 48 h. Conclusions: miR-155 plays a protective role in the progression of atherosclerosis, and it may be achieved by reducing the apoptosis effort of OxLDL to RAW264.7 macrophagocyte.

THE EFFECTS OF PGMS ON IMMUNOLOGICAL FUNCTION IN MICE / 中国海洋药物

The effects of Proplylene glucol mannurate sulfate (PGMS) on mice's immunological function have been studied with immunological techniques. When given orally,PGMS could enhance the weight of the mice's immunological organs and the speed of carbonparticle elimination in mice,the phagocytic rate and index of the macrophagocytes in the mice's abdominal cavity were enhanced, the percentage of ANAE" lymphocytes were increased, the production of hemolysin in mice peripheral blood were promoted. The study suggests that PGMS improves both the specific and nonspecific immunological function in mice.