基于“血水同源”理论探讨清通化瘀汤对脑缺血再灌注损伤模型大鼠脑水肿的影响及相关机制

Effect of Qingtong Huayu Decoction on Cerebral Edema in Rats with Cerebral Ischemia-Reperfusion Injury and Its Related Mechanism Based on the Theory of"Blood and Water Homologation"

目的基于“血水同源”理论探讨清通化瘀汤对脑缺血再灌注损伤模型大鼠脑水肿的影响及相关机制。方法本实验时间为2021年5月。选取180只成年健康雄性SD大鼠,采用随机数字表法将其分为空白组、假手术组、CIRI组、清通化瘀汤组、通心络组,每组36只。空白组大鼠正常饲养,不做任何处理。造模前7 d,假手术组、CIRI组大鼠给予等体积蒸馏水预灌胃,清通化瘀汤组大鼠给予清通化瘀汤2.01 g/kg预灌胃,通心络组大鼠给予通心络0.12 g/kg预灌胃;之后,假手术组、CIRI组、清通化瘀汤组、通心络组大鼠制备高脂血症模型;高脂血症模型制备成功后,CIRI组、清通化瘀汤组、通心络组大鼠制备CIRI模型。造模后次日至取材当天,各组大鼠灌胃方法同前。检测五组大鼠造模后7 d血脂指标(血清TC、TG、LDL-C、HDL-C水平)、神经功能缺损程度(Zea-longa评分),造模后3 h、1 d、3 d、7 d脑含水量,造模后1、3、7 d脑梗死面积,造模后3 h、1 d、3 d、7 d脑组织胶质纤维酸性蛋白(GFAP)、水通道蛋白4(AQP4)、瞬时受体电位香草素受体4型通道(TRPV4)蛋白表达水平。结果造模后7 d,假手术组、CIRI组、清通化瘀汤组、通心络组大鼠血清TC、TG、LDL-C水平高于空白组,血清HDL-C水平低于空白组(P<0.05)。造模后7 d,清通化瘀汤组、通心络组血清TC、TG、LDL-C水平及Zea-longa评分低于CIRI组,血清HDL-C水平高于CIRI组(P<0.05)。造模后3 h、1 d,CIRI组大鼠脑含水量高于空白组(P<0.05);造模后1 d,清通化瘀汤组大鼠脑含水量低于CIRI组(P<0.05)。造模后1、3、7 d,清通化瘀汤组和通心络组大鼠脑梗死面积小于CIRI组(P<0.05)。造模后3 h、1 d、3 d、7 d,CIRI组、清通化瘀汤组、通心络组大鼠脑组织GFAP、AQP4、TRPV4蛋白表达水平高于空白组、假手术组,清通化瘀汤组、通心络组大鼠脑组织GFAP、AQP4、TRPV4蛋白表达水平低于CIRI组,清通化瘀汤组大鼠脑组织GFAP、AQP4、TRPV4蛋白表达水平低于通心络组(P<0.05)。结论清通化瘀汤能有效改善CIRI模型大鼠血脂水平,减轻神经功能缺损及急性期脑水肿程度,缩小脑梗死面积;其减轻脑水肿的机制可能与抑制GFAP、AQP4、TRPV4蛋白表达有关。

Objective To discuss the effect of Qingtong Huayu decoction on cerebral edema in rats with cerebral ischemia-reperfusion injury(CIRI)and its related mechanism based on the theory of"blood and water homologation".Methods The experiment was conducted in May 2021.A total of 180 adult healthy male SD rats were selected and divided into blank group,sham operation group,CIRI group,Qingtong Huayu decoction group and Tongxinluo group by random number table method,with 36 rats in each group.The rats in the blank group were fed normally without any treatment.At 7 days before modeling,the rats in sham operation group and CIRI group were given equal volume of distilled water by pre-gavage,the rats in Qingtong Huayu decoction group were given Qingtong Huayu decoction 2.01 g/kg by pre-gavage,and the rats in Tongxinluo group were given Tongxinluo 0.12 g/kg by pre-gavage.After that,the hyperlipidemia model was prepared in the sham operation group,CIRI group,Qingtong Huayu decoction group and Tongxinluo group.After successful preparation of hyperlipidemia model,CIRI group,Qingtong Huayu decoction group and Tongxinluo group were used to prepare CIRI model.From the next day after modeling to the day of sampling,the gavage method of rats in each group was the same as before.Blood lipid indexes(serum TC,TG,LDL-C,HDL-C levels)and degree of neurological deficit(Zea-longa score)at 7 days after modeling,brain water content at 3 h,1 d,3 d and 7 d after modeling,cerebral infarction area at 1 d,3 d and 7 d after modeling,the expression levels of glial fibrillary acidic protein(GFAP),aquaporin-4(AQP4)and transient receptor potential vanilloid receptor 4(TRPV4)protein of brain tissue at 3 h,1 d,3 d and 7 d after modeling of the five groups were detected.Results At 7 days after modeling,the levels of serum TC,TG and LDL-C in the sham operation group,CIRI group,Qingtong Huayu decoction group and Tongxinluo group were higher than those in the blank group,and the level of serum HDL-C was lower than that in the blank group(P<0.05).At 7 days after modeling,the levels of serum TC,TG and LDL-C and Zea-longa score in Qingtong Huayu decoction group and Tongxinluo group were lower than those in CIRI group,and the level of serum HDL-C was higher than that in CIRI group(P<0.05).At 3 h and 1 d after modeling,the brain water content in CIRI group was higher than that in blank group(P<0.05);at 1 d after modeling,the brain water content in Qingtong Huayu decoction group was lower than that in CIRI group(P<0.05).At 1 d,3 d and 7 d after modeling,the cerebral infarction area in Qingtong Huayu decoction group and Tongxinluo group was smaller than that in CIRI group(P<0.05).At 3 h,1 d,3 d and 7 d after modeling,the expression levels of GFAP,AQP4 and TRPV4 protein of brain tissue in CIRI group,Qingtong Huayu decoction group and Tongxinluo group were higher than those in blank group and sham operation group,and those in Qingtong Huayu decoction group and Tongxinluo group were lower than those in CIRI group,and those in Qingtong Huayu decoction group were lower than those in Tongxinluo group(P<0.05).Conclusion Qingtong Huayu decoction can effectively improve the blood lipid level of CIRI model rats,reduce the degree of neurological deficit,acute cerebral edema and area of cerebral infarction.The mechanism of Qingtong Huayu decoction in reducing brain edema may be related to the inhibition of GFAP,AQP4 and TRPV4 protein expression.