摘要
目的:探讨抗血小板溶栓素(anti-platelet thrombolysin,APT)对大鼠脑缺血再灌注损伤保护作用的机制。方法:采用TLR4-siRNA在体转染技术,下调大鼠脑组织中Toll样受体4(TLR4)蛋白表达。分别取野生型和TLR4下调的SD大鼠,随机分为假手术组,模型组,TLR4阻断剂(TAK-242 2 mg/kg)组,APT高、中、低组(0. 02、0. 01、0. 005 mg/kg),每组8只。线拴法建立大鼠局灶性脑缺血/再灌注损伤模型,G-LISA法测定大鼠脑组织中RhoA蛋白活性,Western blot法测定脑组织中TLR4、RhoA相关卷曲螺旋形成蛋白激酶(ROCK1/2)、磷酸化的c-Jun氨基末端激酶(pJNK)蛋白表达。结果:与对照组比较,TLR4-siRNA在体转染大鼠脑组织中TLR4蛋白表达明显下降;在野生型大鼠中,与模型组比较,抗血小板溶栓素可明显降低脑组织中TLR4蛋白表达,抑制RhoA活性,减少ROCK1/2、p-JNK蛋白表达;在TLR4下调大鼠中,与模型组比较,APT对脑组织中RhoA活性,ROCK1/2、p-JNK蛋白表达无明显影响。结论:抗血小板溶栓素对大鼠脑缺血再灌注损伤保护作用机制主要与其抑制TLR4/RhoA/ROCK信号通路有关。
AIM: To investigate the protective mechanism of anti-platelet thrombolysin( APT) on cerebral ischemia reperfusion injury in rats.METHODS: The TLR4 siRNA in vivo transfection technique was used to reduce the TLR4 expression in the brain of rat. Wild-type and TLR4 down-regulated rats were randomly divided into six groups:sham group,model group,TAK-242( Toll-like receprot 4 blocker) 2 mg/kg group,APT 0. 02 mg/kg,0. 01 mg/kg,0. 005 mg/kg group respectively with eight rats in each group. The rat focal ischemia-reperfusion injury model was established by ligation of middle cerebral artery occlusion( MCAO).RhoA activity was measured using absorbance based G-LISA RhoA activation assay and the expression of TLR4,ROCK1/2 and p-JNK were determined by Western blot. RESULTS: The expression of TLR4 protein in TLR4 siRNA transfected rats brain tissue decreased significantly compared with that in control group rats. Compared with model group,APT could obviously decrease the TLR4,ROCK1/2 and p-JNK protein expression,inhibit the RhoA activity in wild rat brain; however,APT had no significant effect on RhoA activity,ROCK1/2 and p-JNK protein expression in brain compared with model group in TLR4 siRNA transfected rats. CONCLUSION: The protective mechanism of APT on cerebral ischemia reperfusion injury in rats is mainly related to the inhibition of TLR4/RhoA/ROCK signaling pathway.
作者
罗胜勇
贾德武
李小羿
戴向荣
LUO Shengyong;JIA Dewu;LI Xiaoyi;DAI Xiangrong(Anhui Provincial Institute of Medical Science,Hefei 230061,Anhui,China;Affiliated High-tech Hospital of Anhui Medical University,Hefei 230032,Anhui,China;Zhaoke Pharmaceutical (Hefei)Company Limited,Hefei 230088,Anhui,China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2018年第10期1109-1115,共7页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
安徽省自然科学基金资助项目(1608085MH161)
