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范俊逸,陈永斌,刘启华,罗燕萍,李南方,黄显雯.龙蛭汤对大鼠脑缺血再灌注损伤后Caspase-1和IL-18蛋白表达的影响[J].湖南中医药大学学报,2021,41(4):518-522[点击复制] |
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龙蛭汤对大鼠脑缺血再灌注损伤后Caspase-1和IL-18蛋白表达的影响 |
范俊逸,陈永斌,刘启华,罗燕萍,李南方,黄显雯 |
(广西医科大学, 广西 南宁 530021;广西医科大学第一附属医院, 广西 南宁 530021) |
摘要: |
目的 通过使用龙蛭汤对大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型大鼠进行干预,观察脑组织中焦亡相关蛋白Caspase-1和IL-18表达情况,探讨其对脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)的保护作用。方法 将48只SD大鼠随机分为假手术组、模型组、龙蛭汤组、丁苯酞组,共4组,每组12只。除假手术组外,其余3组均采用Longa线栓法建立MCAO大鼠模型。造模成功后丁苯酞组、龙蛭汤组分别予丁苯酞[40 mg/(kg·d)]、龙蛭汤[13.6 g/(kg·d)]灌胃,假手术组、模型组均予等量生理盐水灌胃。术后第3天对各组大鼠行神经功能缺损评分;使用TTC染色法对新鲜脑片进行染色,同时行脑梗死体积测定;HE染色法观察脑皮质区病理形态变化;免疫组化法检测脑组织Caspase-1和IL-18蛋白表达情况。结果 神经功能缺损评分结果显示,假手术组为0分;各造模组评分较假手术组升高,差异有统计学意义(P<0.05)。脑梗死体积百分比结果显示,假手术组无梗死灶;模型组最高,龙蛭汤组、丁苯酞组次之,均较模型组明显降低,差异有统计学意义(P<0.05)。脑组织HE染色结果显示,假手术组几乎未见受损细胞;模型组脑组织皮层神经细胞损伤严重,龙蛭汤组、丁苯酞组均较之有明显改善。免疫组化结果显示:假手术组仅见极少阳性细胞表达;与模型组比较,龙蛭汤组和丁苯酞组的Caspase-1和IL-18蛋白明显降低,差异有统计学意义(P<0.05)。结论 龙蛭汤在改善大鼠CIRI后症状和保护大鼠损伤后神经细胞方面有显著的效果,其机制可能与下调Caspase-1和IL-18蛋白表达有关。 |
关键词: 脑缺血再灌注损伤 龙蛭汤 细胞焦亡 Caspase-1 IL-18 |
DOI:10.3969/j.issn.1674-070X.2021.04.005 |
投稿时间:2021-01-01 |
基金项目:国家自然科学基金项目(81560790);广西中医药重点学科建设项目(GZXK-Z-20-52);陈永斌广西名老中医工作室建设项目。 |
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Effects of Longzhi Decoction on the Expression of Caspase-1 and IL-18 Proteins After Cerebral Ischemia Reperfusion Injury Rats |
FAN Junyi,CHEN Yongbin,LIU Qihua,LUO Yanping,LI Nanfang,HUANG Xianwen |
(Guangxi Medical University, Nanning, Guangxi 530021, China;The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, China) |
Abstract: |
Objective To investigate the expression of Caspase-1 and IL-18 in cerebral artery occlusion (MCAO) model of rats with Longzhi Decoction, and to investigate the protective effect of Longzhi Decoction on cerebral ischemia reperfusion injury (CIRI). Methods 48 SD rats were randomly divided into sham operation group, model group, Longzhi Decoction group and butylphthalide group, with 12 rats in each group. Except for sham operation group, MCAO rat model was established by Longa thread bolting method in the other 3 groups. After successful modeling, butylphthalide group and Longzhi Decoction group were given intragastric gavage of butylphthalide[40 mg/(kg·d)] and Longzhi Decoction[13.6 g/(kg·d)], respectively. Sham operation group and model group were given intragastric gavage of the same amount of normal saline. On the 3rd day after operation, nerve function deficit was scored in each group. TTC staining method was used to stain fresh brain slices and cerebral infarct volume was measured simultaneously. The pathological changes of cerebral cortex were observed by HE staining. The expression of Caspase-1 and IL-18 in brain tissues was detected by immunohistochemistry. Results The neurological deficit score in the sham operation group was 0. The scores of each model group were higher than those of the sham operation group, and the differences were statistically significant (P<0.05). The percentage of cerebral infarction volume showed that there was no infarction in the sham operation group. The highest value was found in model group, followed by Longzhi Decoction group and butylphthalide group, which were significantly lower than model group, with statistical significance (P<0.05). HE staining of brain tissues showed that there were almost no damaged cells in the sham operation group. The damage of cortical nerve cells in brain tissue was serious in model group, but was significantly improved in Longzhi Decoction group and butylphthalide group. Immunohistochemical results showed that only a few positive cells were found in the sham operation group. Compared with model group, the protein levels of Caspase-1 and IL-18 in Longzhi Decoction group and butylphthalide group were significantly decreased, with statistical significances (P<0.05). Conclusion Longzhi Decoction has significant effects on improving symptoms after CIRI and protecting nerve cells after injury in rats, and the mechanism may be related to the down-regulation of Caspase-1 and IL-18 protein expression. |
Key words: cerebral ischemia reperfusion injury Longzhi Decoction pyroptosis Caspase-1 IL-18 |
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