摘要
目的探讨犀角地黄汤对缺血性脑卒中(AIS)再灌注大鼠模型自噬水平的调节机制。方法无特定病原级雄性大鼠60只,随机分为假手术组(A组,只分离颈总动脉不插入线栓)、AIS再灌注组(B组,构建AIS再灌注模型)、AIS再灌注+犀角地黄汤组(C组,于构建AIS再灌注模型前30 min经腹腔注射10 mg/kg犀角地黄汤)、AIS再灌注+犀角地黄汤+JNK抑制剂组(D组,于构建AIS再灌注模型前30 min经腹腔注射10 mg/kg犀角地黄汤和JNK抑制剂),每组各15只。于建模后24 h麻醉大鼠取脑组织做切片,分别进行神经功能评分、自噬超微结构观察、Western Blot检测自噬蛋白Beclin 1和微管相关蛋白轻链3(LC3)表达、TTC测定大鼠脑梗死体积、TUNEL染色法检测细胞凋亡率。结果构建AIS再灌注模型后,与A组大鼠比较,其余组大鼠神经功能均出现异常,自噬泡增加,Beclin 1、LC3蛋白表达增加。与B组比较,C组大鼠神经功能评分、自噬水平、脑梗死体积、Beclin 1、LC3蛋白表达、神经细胞凋亡率降低(P <0. 05);与C组比较,D组大鼠自噬水平显著降低,神经功能进一步改善(P <0. 05)。结论 AIS再灌注后神经细胞损伤严重、自噬水平升高,犀角地黄汤可通过激活JNK通路显著降低自噬水平而降低神经功能损伤,这可能为AIS再灌注后神经损伤提供新的治疗靶点。
Objective To explore the mechanism of Xijiao Dihuang decoction on autophagy in rats with ischemic stroke and reperfusion.Methods Sixty non-specific pathogenic male rats purchased from the Beijing Medical Equipment Inspection Institute( Animal Experimental Center) were randomly divided into a sham operation group( group A,n = 15) and an AIS reperfusion group( group B,to construct AIS reperfusion model,n = 15),B + rhino dihuang decoction group( group C,intraperitoneal injection of Xijiao dihuang decoction on the basis of group B,n = 15),C + JNK inhibitor group( group D,based on group C,an equal volume of JNK inhibitor was injected,n = 15). The brain tissues were taken from the anesthetized rats 24 h after modeling for sectioning,and neurological scores,ultrastructural observation of autophagy,Western Blot were used to detect the expression of autophagy protein Beclin 1 and microtubule-related protein light chain 3( LC3),the volume of cerebral infarction in rats was measured by TTC,and the apoptosis rate was detected by TUNEL staining. Results After constructing the AIS reperfusion model,compared with rats in group A,the neurological function of the other groups was abnormal,autophagic vesicles increased,and Beclin 1,LC3 protein expression increased. Compared with group B,the neurological score,autophagy level,cerebral infarction volume,Beclin 1,LC3 protein expression,and neuronal apoptosis rate in group C rats were reduced( P < 0. 05). Compared with group C,rats in group D Phagocytic levels were significantly reduced and neural function was further improved( P < 0. 05). Conclusion After AIS reperfusion,the nerve cells are severely damaged and the level of autophagy is increased. Xijiao Dihuang Decoction can significantly reduce autophagy and reduce neurological damage by activating the JNK pathway,which may be a new target for nerve injury after AIS reperfusion.
作者
潘思敏
林兴栋
沈志勇
PAN Si-min;LIN Xing-dong;SHEN Zhi-yong(The First Clinical Medical College of Guangzhou University of Traditional Chinese Medicine,Guangzhou Guangdong 510405,China;The Third Affiliated Hospital of Guangzhou University of traditional Chinese medicine,Guangzhou Guangdong 510405,China;Shenzhen Hospital of Shanghai University of Traditional Chinese Medicine,Shenzhen Guangdong 518001,China)
出处
《临床和实验医学杂志》
2020年第4期348-352,共5页
Journal of Clinical and Experimental Medicine
基金
广东省自然科学基金项目(编号:9152402301000006)
关键词
大鼠
缺血性脑卒中再灌注
犀角地黄汤
自噬水平
Rats
Ischemic stroke reperfusion
Xijiao Dihuang Decoction
Autophagy level
