摘要
目的观察抗β2GPⅠ-β2GPⅠ复合物刺激小鼠腹腔巨噬细胞表达TNF-α、IL-1β、IL-6的效果,探讨Toll样受体4(TLR4)在其中的作用。方法对C3H/He N(TLR4正常)小鼠、C3H/He J(TLR4缺陷)小鼠腹腔注射非特异性兔Ig G(R-Ig G)、抗β2GPⅠ抗体,72 h后用细胞免疫荧光法检测小鼠腹腔巨噬细胞TNF-α、IL-1β、IL-6的表达水平;抗β2GPⅠ-β2GPⅠ复合物对小鼠腹腔巨噬细胞进行体外刺激,荧光定量PCR法检测细胞表达上述因子的mRNA水平,western blot法检测其蛋白质分泌水平。结果抗β2GPⅠ抗体刺激的C3H/He N小鼠腹腔巨噬细胞产生TNF-α、IL-1β、IL-6的水平明显高于C3H/He J小鼠,并经细胞免疫荧光法验证;经体外刺激后,荧光定量PCR法结果显示抗β2GPⅠ-β2GPⅠ复合物诱导C3H/He N小鼠腹腔巨噬细胞TNF-α、IL-1β、IL-6 mRNA表达水平高于空白组或C3H/He J刺激组(P均<0.01);western blot结果显示C3H/He N来源细胞TNF-α、IL-1β、IL-6分泌均显著高于C3H/He J来源细胞(P均<0.01)。结论抗β2GPⅠ-β2GPⅠ复合物能够促进小鼠腹腔巨噬细胞炎症因子TNF-α、IL-1β、IL-6的表达,TLR4为介导此过程的受体之一。
Objective To explore the effects of anti-β2-glycoprotein Ⅰ / β2-glycoprotein Ⅰ（ anti-β2GP Ⅰ / β2GP Ⅰ） complex on the expression of TNF-α,IL-1β and IL-6 in mouse peritoneal macrophages and the roles of Toll-like receptor 4（ TLR4） in the process of expression of inflammatory factors. Methods The C3 H / He N mice（ TLR4 intact） and C3 H / He J mice（ TLR4 deficient） were pretreated by intraperitoneal injection with nonspecific rabbit-Ig G（ R-Ig G） and anti-β2GPⅠ antibody respectively. After 72 hours,the expressions of TNF-α,IL-1β and IL-6 in mice peritoneal macrophages were detected by immunofluorescence. The peritoneal macrophages of mouse were stimulated simultaneously with anti-β2GPⅠ / β2GPⅠ complex in vitro. The mRNA levels and the protein expression of above-mentioned factors were tested by real-time PCR and western blot respectively. Results Immunofluorescence analysis showed the expressions of TNF-α,IL-1β and IL-6 in the peritoneal macrophage of C3 H / He N mice stimulated by anti-β2GPⅠ were significantly higher than those of C3 H / He J mice group. In vitro,the mRNA expression levels of TNF-α,IL-1β and IL-6 in C3 H / He N group were also higher than those of C3 H / He J group or blank group（ P〈 0. 01）. Western blot analysis demonstrated that the protein expressions of TNF-α,IL-1β and IL-6 in the peritoneal macrophage of C3 H / He N mice was significantly higher than those of C3 H / He J group.Conclusion Anti-β2GPⅠ / β2GPⅠ complex can enhance TNF-α,IL-1β and IL-6 expression in mouse peritoneal macrophage,which might be mediated by TLR4 signaling pathways.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2014年第12期934-938,共5页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金项目(81370614)
江苏大学学生科研立项(Y13A100)
