七叶皂苷通过抑制TXNIP/MLXIPL信号通路改善脑出血大鼠神经性损伤  

Aescin Ameliorates Neurological Injury in Rats with Cerebral Hemorrhage by Inhibiting TXNIP/MLXIPL Signaling Pathway

作　　者：姬令山[1] 秦合伟[1] 程率芳[1] 赵晶[1] JI Lingshan;QIN Hewei;CHENG Shuaifang;ZHAO Jing(The Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,Henan,China)

机构地区：[1]河南中医药大学第二附属医院,河南郑州450002

出　　处：《辽宁中医杂志》2023年第4期194-198,F0003,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81704030);河南省中医药拔尖人才培养项目(2021-24)。

摘　　要：目的七叶皂苷具有抗炎、消肿和改善血液循环的作用,但其在脑出血治疗中的作用及其机制尚未阐明。探究七叶皂苷对脑出血大鼠神经性功能的影响及潜在的作用机制。方法选取250~280 g雄性Wistar大鼠,采用胶原酶注射法构建大鼠脑出血(Intracerebral haemorrhage,ICH)模型,将其随机分为假手术组(sham)、ICH组、5或10 mg/kg七叶皂苷治疗组。体外,大鼠小胶质细胞HAPI经血红素(Hemin)处理,分为Control组、Hemin组、5或10μM七叶皂苷处理组。脑组织图评估大鼠脑水肿程度,HE染色观察大鼠脑组织的病理学变化,ELISA测定用来评估大鼠氧化应激水平。RT-qPCR和Western blot法分别检测miR-128-3p表达量、硫氧还蛋白相互作用蛋白(Thioredoxin-Interacting Protein,TXNIP)和MLX相互作用蛋白(MLX Interacting Protein-Like,MLXIPL)水平。荧光素报告基因实验验证miR-128-3p和TXNIP的靶定关系。Co-IP验证TXNIP和MLXIPL的相互作用。结果与Sham组大鼠相比,ICH大鼠脑水肿程度增加,HE染色可见神经细胞胞体皱缩且细胞核固缩,血清中炎性因子分泌量和氧化应激产物增加,经七叶皂苷治疗后损伤程度减缓,且10 mg/kg七叶皂苷效果明显优于5 mg/kg七叶皂苷(P<0.05)。TXNIP是miR-128-3p的一个靶基因,且TXNIP可正向调节MLXIPL。ICH大鼠脑组织以及Hemin处理的HAPI细胞中miR-128-3p表达量增加,TXNIP和MLXIPL表达量均降低(P<0.05),且大剂量的七叶皂苷效果优于小剂量七叶皂苷(P<0.05)。结论七叶皂苷通过抑制TXNIP/MLXIPL信号通路,减轻脑水肿,降低氧化应激水平来改善ICH大鼠神经性损伤。Aescin has anti-inflammatory,swelling and improving blood circulation effects,but its role in the treatment of cerebral hemorrhage and its mechanism have not yet been elucidated.Objective To explore the effect of aescin on the neurological function of rats with cerebral hemorrhage and its potential mechanism.Methods Male Wistar rats(weighting 250-280 g)were selected,and an intracerebral haemorrhage(ICH)model was constructed by collagenase injection.The rats were randomly divided into sham operation group(sham),ICH group,5 or 10 mg/kg aescin treatment groups.In vitro,rat microglia HAPI were treated with Hemin and divided into control group,Hemin group,and 5 or 10μM aescin treatment groups.Brain tissue detection was used to assess the degree of brain edema in rats.HE staining was used to observe the pathological changes of the brain tissue and ELISA was used to assess the oxidative stress level in rats.RT-qPCR and Western blot assays were used to detect the expressions of miR-128-3p,the levels of thioredoxin-interacting protein(TXNIP)and MLX interacting protein-like(MLXIPL),respectively.Fluorescein reporter gene experiment verified the targeting relationship between miR-128-3p and TXNIP.Co-IP assay verified the interaction between TXNIP and MLXIPL.Results Compared with rats in the sham group,the degree of brain edema in ICH rats was increased,the cell bodies of nerve cells shrank and the nucleus was shrinkage,and the secretion of inflammatory factors and oxidative stress products in the serum were increased.And the degree of injury was reduced after aescin treatment,and the effect of 10 mg/kg aescin was significantly better than that of 5 mg/kg aescin(P<0.05).TXNIP was a target gene of miR-128-3p,and TXNIP could positively regulate MLXIPL.The expression of miR-128-3p was increased in the brain tissue of ICH rats and HAPI cells treated with Hemin,and the levels of TXNIP and MLXIPL were decreased(P<0.05),and the effect of high-dose aescin was significantly better than that of low-dose aescin(P<0.05).ConclusionAescin

关 键 词：七叶皂苷 脑出血 脑水肿 miR-128-3p TXNIP MLXIPL 

分 类 号：R282.71[医药卫生—中药学]