毒死蜱对大鼠脑组织RAGE介导Gq-PKC信号转导途径影响

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目的探讨48%毒死蜱(CPF)对大鼠胆碱酯酶活力及晚期糖基化终末产物受体(RAGE)介导的Gq-蛋白激酶C(PKC)信号转导途径影响。方法将大鼠随机分为CPF低(1/40LD50)、中(1/20LD50)、高(1/10LD50)3个剂量组和生理盐水对照组,经口染毒28 d后,用三氯化铁比色法检测大鼠全血胆碱酯酶活力;荧光免疫组化法检测脑组织RAGE、PKC的表达,蛋白质印迹法(western blot)检测脑组织中Gq蛋白的表达。结果全血胆碱酯酶活力随染毒剂量的增加而降低,对照组和CPF低、中、高剂量组三氯化铁比色法测定值分别为(33.75±5.03)、(29.70±4.01)、(10.40±1.95)、(5.25±0.05);RAGE、Gq蛋白、PKC的表达均随染毒剂量的增加而增高,对照组和CPF低、中、高剂量组RAGE免疫反应平均光密度值分别为(0.32±0.01)、(2.01±0.09)、(5.03±0.05)、(8.02±0.04),Gq蛋白的相对含量分别为(0.84±0.01)、(0.90±0.01)、(1.10±0.02)、(1.59±0.01),PKC免疫反应平均光密度值分别为(0.12±0.01)、(2.01±0.03)、(3.52±0.02)、(5.01±0.08)。结论 CPF主要通过抑制胆碱脂酶的活力对机体造成损伤,RAGE、Gq蛋白、PKC的高表达可能参与了CPF中毒神经系统病理性信号转导。 Objective To investigate the effect of 48% chlorpyrifos (CPF) on the activity of cholinesterase and the receptor for advanced glycation end products (RAGE) mediated Gq-protein kinase C (PKC) signal transduction pathway in rats. Methods The rats were randomly divided into 3 groups (1 / 40LD50, 1 / 20LD50, 1 / 10LD50) and saline control group. After oral administration for 28 days, The activity of cholinesterase in rat whole blood was detected by iron colorimetric method. The expression of RAGE and PKC in brain tissue was detected by fluorescence immunohistochemistry. The expression of Gq protein in brain tissue was detected by western blot. Results The whole blood cholinesterase activity decreased with the increase of the dose. The values ​​of the three ferric chloride methods in the control group and the low, medium and high dose CPF groups were (33.75 ± 5.03), (29.70 ± 4.01) , (10.40 ± 1.95) and (5.25 ± 0.05) respectively. The expression of RAGE, Gq and PKC increased with the increase of exposure dose. The mean optical density of RAGE immunoreactivity in control group and low, middle and high dose CPF group were (0.84 ± 0.01), (0.90 ± 0.01) and (1.10 ± 0.02), respectively, and the relative content of Gq protein were (0.32 ± 0.01), (2.01 ± 0.09), (5.03 ± 0.05) and (8.02 ± 0.04) (1.59 ± 0.01). The average optical density of PKC immunoreactivity were (0.12 ± 0.01), (2.01 ± 0.03), (3.52 ± 0.02) and (5.01 ± 0.08) respectively. Conclusion CPF mainly damages the body through inhibiting the activity of cholinesterase. The high expression of RAGE, Gq protein and PKC may be involved in the pathological signal transduction of the nervous system of CPF intoxication.
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