脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

Objective To compare the morphologic change and cytokines expression in RAW264.7 and Ana-1 stimulated by lipopolysaccharide(LPS). Methods RAW264.7 and Ana-1 were cultivated with various concentrations of LPS(0.1 mg/L, 1 mg/L, 10 mg/L, 100 mg/L). MTT was performed to evaluate the proliferation ability of cells. Two kinds of cells were cultivated with 1 mg/L. Then,the concentrations of TNF-α,IL-1β,IL-6,IL-10 in culture medium were detected by ELISA in different times(0 h,4 h,8 h,12 h,24 h). Results The survival rates of RAW264.7 and Ana-1 (162.05±28.14)% and (159.92±20.43)% were significantly higher in 1 mg/L group than those of other groups. When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 expressed cytokines in time-dependent manner (increased first and decreased finally). The concentrations of TNF-α and IL-10 in RAW264.7 group were higher than those Ana-1 group in 4 h only. IL-1β and IL-6,however,were in higher concentrations in RAW264.7 group than Ana-1 group all the time. Conclusions When stimulated with 1 mg/L LPS, both RAW264.7 and Ana-1 have higher survival rate. RAW264.7 and Ana-1 have different ability in expressing cytokines after being stimulated by LPS. Therefore,RAW264.7 and Ana-1 have variant response to LPS slightly.     

β-arrestin 2 attenuates lipopolysaccharide-induced liver injury via inhibition of TLR4/NF-κB signaling pathwaymediated inflammation in mice

AIM To study the role and the possible mechanism of β-arrestin 2 in lipopolysaccharide(LPS)-induced liver injury in vivo and in vitro.METHODS Male β-arrestin 2~(+/+) and β-arrestin 2~(-/-)C57 BL/6 J mice were used for in vivo experiments, and the mouse macrophage cell line RAW264.7 was used for in vitro experiments. The animal model was established via intraperitoneal injection of LPS or physiological sodium chloride solution. Blood samples and liver tissues were collected to analyze liver injury and levels of pro-inflammatory cytokines. Cultured cell extracts were collected to analyze the production of pro-inflammatory cytokines and expression of key molecules involved in the TLR4/NF-κB signaling pathway.RESULTS Compared with wild-type mice, the β-arrestin 2 knockout mice displayed more severe LPS-induced liver injury and significantly higher levels of proinflammatory cytokines, including interleukin(IL)-1β, IL-6, tumor necrosis factor(TNF)-α, and IL-10. Compared with the control group, pro-inflammatory cytokines(including IL-1β, IL-6, TNF-α, and IL-10) produced by RAW264.7 cells in the β-arrestin 2 si RNA group were significantly increased at 6 h after treatment with LPS. Further, key molecules involved in the TLR4/NF-κB signaling pathway, including phosphoIκBα and phosho-p65, were upregulated.CONCLUSION β-arrestin 2 can protect liver tissue from LPS-induced injury via inhibition of TLR4/NF-κB signaling pathwaymediated inflammation.     

脂多糖刺激RAW264.7和Ana-1形态改变及细胞因子表达的差异

目的 比较两株小鼠来源的巨噬细胞株RAW264.7和Ana-1经脂多糖(lipopolysaccharide,LPS)诱导后在形态及细胞因子表达等方面的差异.方法 MTT法检测小同终浓度(0.1 mg/L、1 mg/L、10 mg/L、100 mg/L)LPS作用后两株细胞增殖活力,确定最佳作用浓度.选择1 mg/L LPS刺激RAW264.7和Ana-1细胞,ELISA法分别于0 h、4 h、8 h、12 h、24 h检测两株细胞肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、IL-6和IL-10的表达量.结果 LPS终浓度为1 mg/L时两株细胞存活率分别为(162.05±28.14)%、(159.92±20.43)%,显著大于同细胞其他浓度组别(P<0.01);1 mg/L LPS刺激后,两株细胞各细胞因子表达均为随时间呈先增多后减少趋势,峰值出现时间RAW264.7细胞早于Ana-1细胞.TNF-α的表达4 h时RAW264.7细胞高于Ana-1(P<0.01),8 h、12 h时低于后者(P<0.05);IL-1β的表达各时间点比较RAW264.7均高于Ana-1(P<0.05);IL-6的表达各时间点比较RAW264.7均低于Ana-1(P<0.05);IL-10的表达于刺激后4 h,RAW264.7细胞高于Ana-1细胞(P<0.05).结论 当LPS浓度为1 mg/L时,Ana-1与RAW264.7细胞存活率均最强;经LPS刺激后,RAW264.7细胞TNF-α、IL-6、IL-1β、IL-10表达高峰早于Ana-1细胞,各细胞因子表达量各有侧重.研究者进行炎症相关实验时对Ana-1和RAW264.7的选择需考虑两者之间的差异.