汉黄芩素通过调控JNK/AP-1信号通路对缺血性脑卒中大鼠神经功能障碍、脑组织损伤的影响

Effects of wogonin on neurological dysfunction and brain tissue injury in a rat model of ischemic stroke via regulation of JNK/AP-1 signaling pathway

目的探讨汉黄芩素通过调控c-Jun氨基末端激酶(JNK)/激活蛋白-1(AP-1)信号通路对缺血性脑卒中(CIS)大鼠神经功能障碍及脑组织损伤的影响。方法大鼠随机分为对照组、模型组、汉黄芩素低剂量组(0.07 mg/kg)、汉黄芩素高剂量组(0.14 mg/kg)、丁苯酞组(20 mg/kg)、汉黄芩素高剂量+JNK激活剂(Anisomycin)组(0.14 mg/kg+5 mg/kg),每组20只。对照组大鼠仅切开皮肤、分离血管,不进行拴线处理,其余各组均按照改良的线栓法构建CIS模型。建模成功后,各组给予相应剂量药物,每天1次,连续14 d。Zea-Longa评分法评价大鼠神经功能,TTC染色测定大鼠脑梗死体积,干湿重法测定大鼠脑组织含水量,HE染色观察大鼠脑组织海马CA1区神经细胞病理变化,试剂盒检测大鼠脑组织超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平,TUNEL染色检测大鼠脑组织神经细胞凋亡,Western blot法检测大鼠脑组织p-JNK1、p-c-Jun、p-c-Fos蛋白表达。结果与对照组比较,模型组大鼠海马CA1区神经细胞病理损伤严重,神经功能评分、脑梗死体积百分比、脑组织含水量、MDA水平、神经细胞凋亡率、p-JNK1、p-c-Jun、p-c-Fos蛋白表达升高(P<0.05),SOD活性降低(P<0.05);与模型组比较,汉黄芩素各剂量组和丁苯酞组以上指标改善(P<0.05);Anisomycin减弱了高剂量汉黄芩素对CIS大鼠神经功能障碍及脑组织损伤的改善作用。结论汉黄芩素可能通过抑制JNK/AP-1信号通路来改善CIS大鼠神经功能障碍及脑组织损伤。

AIM To investigate the impacts of wogonin on neurological dysfunction and brain tissue damage in a rat model of ischemic stroke(CIS)via c-Jun N-terminal kinase(JNK)/activator protein-1(AP-1)signaling pathway.METHODS The rats were randomly divided into the control group,the model group,the low-dose wogonin group(0.07 mg/kg),the high-dose wogonin group(0.14 mg/kg),the butylphthalide group(20 mg/kg),the high-dose wogonin(0.14 mg/kg)+JNK activator(Anisomycin)group(5 mg/kg),with 20 rats in each group.In contrast to the rats of the control group only underwent skin incision and blood vessel separation without tethering,the rats of the other groups were induced into CIS models according to the modified suture method,followed by the corresponding drugs administration on a once daily basis for 14 days.The rats had their neurological function assessed using Zea-Longa scoring system;their volume of cerebral infarction determined by TTC staining;their water content in brain tissue detected by wet-dry weight method;their pathological changes of neurons in the hippocampal CA1 region observed by HE staining;their activity of superoxide dismutase(SOD),level of malondialdehyde(MDA)in brain tissue detected using the kits;their neuronal apoptosis in brain tissue investigated by TUNEL staining;their expressions of p-JNK1,p-c-Jun,and p-c-Fos proteins in brain tissue measured by Western blot.RESULTS Compared with the control group,the model group displayed severely damaged neuronal cells in the hippocampal CA1 area;increased neurological function score,cerebral infarction volume percentage,brain tissue water content,MDA level,neuronal apoptosis rate,and expressions of p-JNK1,p-c-Jun,and p-c-Fos proteins(P<0.05);and decreased SOD activity(P<0.05).Compared with the model group,the each dose wogonin group and butylphthalide group performed better in terms of the improvement in all of the aforementioned indexes levels(P<0.05).Anisomycin offset the ameliorating effects of high-dose wogonin on neurological dysfunction and brain tissue damage in CIS rats.CONCLUSION Wogonin may improve the neurological dysfunction and brain tissue damage in CIS rats via JNK/AP-1 pathway.