论文部分内容阅读
目的探讨类固醇受体辅活化子(SRC)-3蛋白在脂多糖诱导小鼠腹腔巨噬细胞趋化和吞噬功能中的作用。方法健康SPF级雌性野生型(SRC-3+/+)小鼠、SRC-3基因敲除(SRC-3-/-)小鼠各5只,分别分离和原代培养腹腔巨噬细胞,并相应分为SRC-3+/+组和SRC-3-/-组。收集并调整细胞浓度为1×106/ml,接种于6孔板,1 ml/孔,给予10μg/ml LPS刺激,分别在刺激前(0 h)、刺激后4 h和12 h收集腹腔巨噬细胞。通过Western blot测定腹腔巨噬细胞Toll样受体4(TLR4)的表达,并通过transwell趋化实验和中性红吞噬实验,分别测定腹腔巨噬细胞的趋化指数(CI)和吞噬功能。结果 LPS诱导前两组腹腔巨噬细胞TLR4的蛋白表达和CI差别无统计学意义,但SRC-3-/-组的吞噬功能显著低于SRC-3+/+组(P<0.01);无论是SRC-3+/+组,还是SRC-3-/-组,LPS诱导4 h和12 h后,两组腹腔巨噬细胞TLR4的表达和CI均显著增高(P<0.01),但在相应时间点,SRC-3-/-组与SRC-3+/+组相比差别无统计学意义。LPS刺激4 h后,两组吞噬功能均显著增加(P<0.01),但SRC-3-/-组增加的程度显著低于SRC-3+/+组(P<0.01);相反,LPS刺激12 h后,两组吞噬功能均受到不同程度的抑制(P<0.05或P<0.01),且SRC-3-/-组较SRC-3+/+组抑制的程度更低(P<0.01)。结论 SRC-3调节腹腔巨噬细胞的先天性免疫功能可能与吞噬功能有关,而与其趋化功能无关。
Objective To investigate the role of steroid receptor co-activator (SRC) -3 in chemotaxis and phagocytosis of murine peritoneal macrophages induced by lipopolysaccharide. Methods Peritoneal macrophages were isolated and primary cultures of healthy SPF female wild type (SRC-3 + / +) mice and SRC-3 knockout (SRC-3 - / - It was divided into SRC-3 + / + group and SRC-3 - / - group. The cells were collected and adjusted to 1 × 106 / ml. The cells were inoculated into a 6-well plate, 1 ml / well and stimulated with 10 μg / ml LPS. Abdominal macrophages were collected before stimulation (0 h), 4 h and 12 h cell. The expression of Toll-like receptor 4 (TLR4) in peritoneal macrophages was detected by Western blot. The chemotactic index (CI) and phagocytic function of peritoneal macrophages were determined by transwell chemotaxis assay and neutral red-phagocytosis assay respectively. Results There was no significant difference in TLR4 protein expression and CI between two groups before LPS induction, but the phagocytic function of SRC-3 - / - group was significantly lower than that of SRC-3 + / + group (P <0.01) (P <0.01). However, in the SRC-3 + / + and SRC-3 - / - groups, the expression of TLR4 and the CI of the two groups of macrophages were significantly increased at 4 h and 12 h after LPS induction At the time point, the difference between SRC-3 - / - group and SRC-3 + / + group was not statistically significant. The phagocytic function of both groups increased significantly after 4 h of LPS stimulation (P <0.01), but the increase of SRC-3 - / - group was significantly lower than that of SRC-3 + / + group After 12 hours, the phagocytic function of both groups was inhibited to some extent (P <0.05 or P <0.01), and the degree of inhibition was lower in SRC-3 - / - group than that in SRC-3 + / + . CONCLUSIONS: SRC-3 regulates innate immune function of peritoneal macrophages, which may be related to phagocytosis but not chemotaxis.