目的探讨蜕皮甾酮(EDS)对脂多糖(LPS)诱导的兔软骨细胞损伤的保护作用及其机制。方法体外分离、培养兔关节软骨细胞,随机分为对照组、LPS诱导损伤组(LPS组),蜕皮甾酮干预组(LPS+EDS组)。MTT法和流式细胞术分别检测各组细胞增殖率及细胞...目的探讨蜕皮甾酮(EDS)对脂多糖(LPS)诱导的兔软骨细胞损伤的保护作用及其机制。方法体外分离、培养兔关节软骨细胞,随机分为对照组、LPS诱导损伤组(LPS组),蜕皮甾酮干预组(LPS+EDS组)。MTT法和流式细胞术分别检测各组细胞增殖率及细胞凋亡率;RT-PCR和Western blot检测软骨细胞中诱导型一氧化氮合酶(i NOS)表达;硝酸还原酶法和ELISA法分别检测各组NO及白细胞介素(IL)-1β含量。结果与对照组相比,LPS组细胞增殖率降低,凋亡率升高,i NOS m RNA和蛋白表达量以及NO和IL-1β含量增高(均P<0.05)。LPS+EDS组较LPS组细胞增殖率升高,凋亡率降低,i NOS m RNA和蛋白表达量及NO和IL-1β的含量降低(均P<0.05)。结论蜕皮甾酮对LPS诱导的兔软骨细胞损伤具有保护作用,其保护作用可能与抑制i NOS表达有关。展开更多
目的：应用大鼠血清建立大鼠体外软骨细胞退变模型。方法：取出生24 h SD大鼠关节处软骨，Ⅱ型胶原酶多次消化后获得原代软骨细胞，取原代细胞进行实验。软骨细胞分别用含10％胎牛血清的DMEM （对照组）、含50 ng／mL 白细胞介素（ IL ...目的：应用大鼠血清建立大鼠体外软骨细胞退变模型。方法：取出生24 h SD大鼠关节处软骨，Ⅱ型胶原酶多次消化后获得原代软骨细胞，取原代细胞进行实验。软骨细胞分别用含10％胎牛血清的DMEM （对照组）、含50 ng／mL 白细胞介素（ IL ）－1β＋10％胎牛血清的 DMEM （ IL－1β组）、含2．5％大鼠血清的DMEM（2．5％血清组）及含5．0％大鼠血清的DMEM（5．0％血清组）中培养。培养24 h后，观察细胞形态变化，MTT法检测各组细胞的增殖情况，蛋白质印迹法检测增殖细胞核抗原的表达和Ⅱ型胶原及MMP－13的表达，实时定量PCR检测退变相关基因ADAMTS5、MMP－9、Aggrecan和SOX－9的表达。结果：两血清组和IL－1β组的软骨形态均由原来的多角形变成长梭形，且两血清组和IL－1β组均促进软骨细胞的增殖，下调转录因子SOX－9和上调基质降解酶MMP－13、MMP－9、ADAMTS5的表达。结论：大鼠血清具有促进软骨退变的作用，可用于建立体外软骨退变模型。展开更多
Chronic activation of microglial cells endangers neuronal survival through the release of various proinflammatory and neurotoxic factors. The root of Paeonia lactiflora Pall has been considered useful for the treatmen...Chronic activation of microglial cells endangers neuronal survival through the release of various proinflammatory and neurotoxic factors. The root of Paeonia lactiflora Pall has been considered useful for the treatment of various disorders in traditional oriental medicine. Paeonol, found in the root of Paeonia lactiflora Pall, has a wide range of pharmacological functions, including anti-oxidative, anti-inflammatory and neuroprotective activities. The objective of this study was to examine the efficacy of paeonol in the repression of inflammation-induced neurotoxicity and microglial cell activation. Organotypic hippocampal slice cultures and primary microglial cells from rat brain were stimulated with bacterial lipopolysaccharide. Paeonol pretreatment was performed for 30 minutes prior to lipopolysaccharide addition. Cell viability and nitrite （the production of nitric oxide）, tumor necrosis factor-alpha and interleukin-lbeta products were measured after lipopolysaccharide treatment. In organotypic hippocampal slice cultures, paeonol blocked lipopolysaccharide-related hippocampal cell death and inhibited the release of nitrite and interleukin-lbeta. Paeonol was effective in inhibiting nitric oxide release from primary microglial cells. It also reduced the lipopolysaccharide-stimulated release of tumor necrosis factor-alpha and intefleukin-1β from microglial cells. Paeonol possesses neuroprotective activity in a model of inflammation-induced neurotoxicity and reduces the release of neurotoxic and proinflammatory factors in activated microglial cells.展开更多
文摘目的探讨蜕皮甾酮(EDS)对脂多糖(LPS)诱导的兔软骨细胞损伤的保护作用及其机制。方法体外分离、培养兔关节软骨细胞,随机分为对照组、LPS诱导损伤组(LPS组),蜕皮甾酮干预组(LPS+EDS组)。MTT法和流式细胞术分别检测各组细胞增殖率及细胞凋亡率;RT-PCR和Western blot检测软骨细胞中诱导型一氧化氮合酶(i NOS)表达;硝酸还原酶法和ELISA法分别检测各组NO及白细胞介素(IL)-1β含量。结果与对照组相比,LPS组细胞增殖率降低,凋亡率升高,i NOS m RNA和蛋白表达量以及NO和IL-1β含量增高(均P<0.05)。LPS+EDS组较LPS组细胞增殖率升高,凋亡率降低,i NOS m RNA和蛋白表达量及NO和IL-1β的含量降低(均P<0.05)。结论蜕皮甾酮对LPS诱导的兔软骨细胞损伤具有保护作用,其保护作用可能与抑制i NOS表达有关。
文摘Chronic activation of microglial cells endangers neuronal survival through the release of various proinflammatory and neurotoxic factors. The root of Paeonia lactiflora Pall has been considered useful for the treatment of various disorders in traditional oriental medicine. Paeonol, found in the root of Paeonia lactiflora Pall, has a wide range of pharmacological functions, including anti-oxidative, anti-inflammatory and neuroprotective activities. The objective of this study was to examine the efficacy of paeonol in the repression of inflammation-induced neurotoxicity and microglial cell activation. Organotypic hippocampal slice cultures and primary microglial cells from rat brain were stimulated with bacterial lipopolysaccharide. Paeonol pretreatment was performed for 30 minutes prior to lipopolysaccharide addition. Cell viability and nitrite （the production of nitric oxide）, tumor necrosis factor-alpha and interleukin-lbeta products were measured after lipopolysaccharide treatment. In organotypic hippocampal slice cultures, paeonol blocked lipopolysaccharide-related hippocampal cell death and inhibited the release of nitrite and interleukin-lbeta. Paeonol was effective in inhibiting nitric oxide release from primary microglial cells. It also reduced the lipopolysaccharide-stimulated release of tumor necrosis factor-alpha and intefleukin-1β from microglial cells. Paeonol possesses neuroprotective activity in a model of inflammation-induced neurotoxicity and reduces the release of neurotoxic and proinflammatory factors in activated microglial cells.
