补阳还五汤通过调控miR-210减轻大鼠脑缺血再灌注损伤的实验研究

Experimental study on reducing cerebral ischemia-reperfusion injury in rats by regulating miR-210 with Buyang Huanwu decoction

目的:探究补阳还五汤是否可通过抑制微小RNA-210(miR-210)靶向调控大脑皮质区B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、胱天蛋白酶3(Caspase-3),减轻大鼠脑缺血再灌注损伤(CIRI)。方法:随机将100只SD大鼠分为五组。模型组和补阳还五汤各组均建立局灶性脑缺血再灌注(MCAO)模型,假手术组做假手术处理。补阳还五汤低、中和高剂量组分别以200、300、400 mg/kg的补阳还五汤汤剂进行灌胃,假手术组和模型组以同体积0.9%氯化钠溶液灌胃,持续灌注7 d。比较各组神经功能缺损评分、脑梗死率、脑含水量和脑皮质区miR-210、Bax、Bcl-2、Caspase-3表达量。结果:灌胃1 d后,补阳还五汤高剂量组与模型组比较,神经功能缺损评分明显下降(P<0.05)。灌胃3 d后,中、高剂量组与模型组和低剂量组,高剂量组与中剂量组比较,其神经功能缺损评分明显下降(P<0.05)。灌胃7 d后,模型组与补阳还五汤三组间比较,补阳还五汤三组之间比较,其神经功能缺损评分、脑梗死率、脑含水量、miR-210、Bax、Bcl-2、Caspase-3间均存在统计学差异(P<0.01)。结论:补阳还五汤可通过抑制miR-210,调节脑皮质区促细胞凋亡/抑细胞凋亡基因表达,从而减轻CIRI大鼠神经功能损伤,且以最高剂量的药效最优。

Objective:To investigate whether Buyang Huanwu decoction can regulate B lymphoblastoma-2-associated X protein(Bax),B lymphoblastoma-2(Bcl-2)and Caspase-3 in cerebral cortex by inhibiting micro RNA-210(miR-210),thereby alleviating ischemia reperfusion injury(CIRI)in rats.Methods:100 SD rats were randomly divided into 5 groups,and focal cerebral ischemia reperfusion(MCAO)model was established in model group and Buyang Huanwu decoration group,sham operation was performed in sham operation group.The low,middle and high dose groups of Buyang Huanwu decoction were given 200 mg/kg,300 mg/kg and 400 mg/kg of Buyang Huanwu decoction,respectively,and the sham operation group and model group were given the same volume of normal saline for continuous perfusion for 7 days.Neurological deficit score,cerebral infarction rate,cerebral water content and expression of miR-210,Bax,Bcl-2 and Caspases-3 in cerebral cortex were compared among all groups.Results:Compared with the model group,the neurological deficit score of high-dose Buyang Huanwu decoration group was decreased after 1 day of intragastration(P<0.05).After 3 days of intragastric administration,compared with model group and low dose group,the neurological deficit score of medium and high dose groups decreased,and compared with model group,high dose group’s neurological deficit scores also decreased(P<0.05).After 7 days of inadministration,there were statistical significance in neural function deficit score,cerebral infarction rate,cerebral water content,miR-210,Bax,Bcl-2 and Caspase-3 between the model group and Buyang Huanwu decoration groups,and between Buyang Huanwu decoration group and Buyang Huanwu Tang group(P<0.01).Conclusion:Buyang Huanwu decoction can regulate the expression of apoptosis-promoting/apoptosis-suppressing genes in the cerebral cortex by inhibiting miR-210,thus alleviating the neurological damage in CIRI rats,and the highest dose has the best efficacy.