摘要
目的 探讨芦丁对慢性脑低灌注大鼠海马组织神经元损伤及Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋蛋白激酶(ROCK)信号通路的影响。方法 将SD大鼠随机分为假手术组、模型组、芦丁低剂量组(20 mg/kg)、芦丁中剂量组(40 mg/kg)、芦丁高剂量组(80 mg/kg)、RhoA抑制剂组(Rhosin hydrochloride,40 mg/kg),每组9只。除假手术组外,其余各组大鼠均通过结扎颈动脉构建慢性脑低灌注大鼠模型,按照各组给药剂量进行干预处理,每天给药1次,持续4周。采用Morris水迷宫实验测定大鼠的认知功能和记忆能力,苏木素-伊红染色观察大鼠海马组织神经元状态,TUNEL染色检测大鼠海马组织神经元凋亡情况,透射电镜观察大鼠海马组织神经元超微结构,蛋白免疫印迹法检测大鼠海马组织凋亡相关蛋白[胱天蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2相关X蛋白(Bax)]及RhoA/ROCK通路相关蛋白表达情况。结果 与假手术组相比,模型组大鼠海马组织神经元数量减少,神经元超微结构、细胞器结构严重受损,逃避潜伏期、海马组织神经元凋亡率、凋亡相关蛋白(Caspase-3、Bax)、RhoA、ROCK1、ROCK2蛋白表达显著升高(P<0.05),穿越平台次数显著减少(P<0.05);与模型组相比,芦丁各剂量组及RhoA抑制剂组大鼠神经元数量显著增多,神经元超微结构、细胞器受损等均得到一定的恢复,逃避潜伏期、海马组织神经元凋亡率、凋亡相关蛋白(Caspase-3、Bax)、RhoA、ROCK1、ROCK2蛋白表达显著降低(P<0.05),穿越平台次数显著增多(P<0.05),且芦丁各剂量组呈剂量依赖效应;芦丁高剂量组与RhoA抑制剂组上述指标比较差异均无统计学意义。结论芦丁可能通过抑制RhoA/ROCK信号通路缓解慢性脑低灌注引起的大鼠神经元损伤。
Objective To investigate the effects of rutin on hippocampal neuron injury and Ras homolog gene family member A/Rho-associated coiled-coil protein kinase(RhoA/ROCK) signaling pathway in chronic cerebral hypoperfusion rats.Methods SD rats were randomly divided into the sham operation group,the model group,the rutin low dose group(20 mg/kg),the rutin medium dose group(40 mg/kg),the rutin high dose group(80 mg/kg) and RhoA inhibitor group(Rhosin hydrochloride,40 mg/kg),with 9 rats in each group.Except the sham operation group,rats in the other groups were subjected to chronic cerebral hypoperfusion by ligating carotid artery.Intervention treatment was carried out according to the dosage of each group.The drug was administered once a day for 4 weeks.Morris water maze test was used to determine the cognitive function and memory ability of rats.Hematoxylin eosin staining was used to observe the status of hippocampal neurons.TUNEL staining was used to detect the apoptosis of hippocampal neurons.Transmission electron microscope was used to observe the ultrastructure of hippocampal neurons.Western blot assay was used to detect the expression levels of apoptosis related proteins [Caspase-3,B cell lymphoma-2 associated X protein(Bax)] and RhoA/ROCK pathway related proteins in hippocampus of rats in each group.Results Compared with the sham operation group,the number of neurons in hippocampus decreased in the model group,the ultrastructure and organelle structure of neurons were seriously damaged,the escape latency,apoptosis rate of hippocampal neurons,expression levels of apoptosis related proteins(Caspase-3,Bax),RhoA,ROCK1 and ROCK2 were significantly increased(P 0.05).Conclusion Rutin can alleviate the neuronal injury induced by chronic cerebral hypoperfusion in rats,which may be achieved by inhibiting RhoA/ROCK signaling pathway.
作者
李艳
刘琼
黄萱
刘丽江
LI Yan;LIU Qiong;HUANG Xuan;LIU Lijiang(Department of Pathology and Pathophysiology,Medical College of Jianghan University,Wuhan 430100,China;Department of Pathology,Affiliated Hospital of Jianghan University(Wuhan Sixth Hospital))
出处
《天津医药》
CAS
北大核心
2022年第12期1270-1276,共7页
Tianjin Medical Journal
基金
湖北省中医药中西医结合科研项目(鄂卫生计生通[2017]20号-11)。
