趋化因子CCL2对小鼠血小板活化影响研究

Effect of chemokine CCL2 on platelet activation in mice

目的探讨趋化因子CCL2对小鼠血小板聚集、颗粒分泌及信号通路分子的影响。方法选取8～10周龄野生型C57雄性小鼠和CCL2^(-/-)雄性小鼠各10只为研究对象。经不同浓度二磷酸腺苷(ADP)及胶原刺激后,采用光比浊法检测小鼠血小板聚集率的差异;酶联免疫吸附法检测小鼠血小板颗粒分泌的白细胞分化抗原40配体(CD40L)及血小板第四因子(PF4)的差异;采用Western Blot检测ADP刺激后小鼠血小板内P38丝裂原活化蛋白激酶(P38MAPK)和热休克蛋白27(HSP27)表达。结果不同浓度ADP及胶原体外刺激后,CCL2^(-/-)组小鼠血小板聚集率、CD40L及PF4均显著低于野生型C57组小鼠,两组比较,差异均有统计学意义(P<0.01)。经ADP刺激的野生型C57组小鼠P38MAPK(T180/Y182)、HSP27(S78/S82)的磷酸化表达水平较刺激前明显升高,差异有统计学意义(P<0.05)。经ADP体外刺激的CCL2^(-/-)组小鼠血小板内P38MAPK(T180/Y182)、HSP27(S78/S82)的磷酸化表达明显低于野生型C57组小鼠,两组比较,差异均有统计学意义(P<0.05)。结论趋化因子CCL2可以作为一种崭新的调控因子参与小鼠血小板聚集、颗粒分泌及信号通路分子P38MAPK(T180/Y182)、HSP27(S78/S82)的活化。

Objective To investigate the effect of chemokine CCL2 on platelet aggregation rate,granule secretion and signal pathway molecules in mice.Methods Ten wild type C57 male mice and 10 CCL2-/-male mice with 8-10 weeks age were selected.Stimulated by different concentration of adenosine diphosphate(ADP)and collagen,using light turbidimetric method to detect the difference of platelet aggregation rate in mice and enzyme-linked immunosorbent method to detect the mice platelet granular secretion of leukocyte differentiation antigen 40 ligand(CD40 L)and platelet factor 4(PF4).Mitogen-activated protein kinase(P38 MAPK)and heat shock protein 27(HSP27)in platelets were tested by Western Blot after stimulation with ADP.Results After in vitro stimulation of ADP and collagen at different concentrations,the platelet aggregation rate,CD40 L and PF4 in CCL2-/-group were significantly lower than those in wild-type C57 group(P<0.01).The phosphorylation levels of P38 MAPK(T180/Y182)and HSP27(S78/S82)in wild-type C57 mice stimulated by ADP were significantly higher than those before the stimulation(P<0.01).The phosphorylation expressions of P38 MAPK(T180/Y182)and HSP27(S78/S82)in the platelets of CCL2-/-group stimulated by ADP in vitro were significantly lower than those of the wild-type C57 group,and the differences were statistically significant(P<0.05).Conclusion As a novel regulatory factor,chemokine CCL2 can be involved in the activation of mouse platelet aggregation rate,particle secretion and signal pathway molecules P38 MAPK(T180/Y182)and HSP27(S78/S82).