破血化瘀、填精补髓法调节实验性Nrf2^(-/-)小鼠脑出血细胞凋亡的研究

Study on the regulation of apoptosis in experimental Nrf2^(-/-) intracerebral hemorrhage mice by activating blood and removing stasis and supplementing essence and marrow

目的观察破血化瘀、填精补髓法通过调节Nrf2对实验性脑出血小鼠细胞凋亡的影响。方法取野生型C57BL/6小鼠和Nrf2^(-/-)小鼠各30只,随机分成6组,每组10只,分别为野生型假手术组,野生型模型组,野生型方剂组,Nrf2^(-/-)假手术组,Nrf2^(-/-)模型组,Nrf2^(-/-)方剂组。采用自体血注射法制作脑出血小鼠模型。方剂组给予破血化瘀、填精补髓中药汤剂灌胃,模型组和假手术组分别以等量生理盐水灌胃。应用Longa评分检测小鼠神经功能,免疫组化法检测Caspase-3、Bax、Bcl-2表达,Western blot法检测Caspase-3、Bax、Bcl-2的蛋白表达。结果与野生型小鼠比较,Nrf2^(-/-)小鼠神经功能减弱(P<0.05),与Nrf2^(-/-)小鼠比较,方剂组小鼠在第三天神经功能有所改善(P<0.05)。免疫组化法和Western blot法结果显示,Nrf2^(-/-)小鼠脑出血凋亡基因Caspase-3和Bax表达增多(P<0.05)并且抑制了抗细胞凋亡因子Bcl-2(P<0.05)的表达,加重了细胞凋亡。结论破血化瘀、填精补髓中药汤剂可能是通过调节Nrf2改善实验性脑出血小鼠细胞凋亡程度。

Objective To observe the effect of Chinese herbal decoction for activating blood and removing stasis and supplementing essence and marrow by regulating Nrf2 on cell apoptosis in mice with experimental intracerebral hemorrhage.Methods Randomly divide 30 C57BL/6 wild-type mice and 30 Nrf2 knockout mice into six groups,they are wild type sham operation group,wild type model group,wild type prescription group,Nrf2^(-/-)sham operation group,Nrf2^(-/-)model group,Nrf2^(-/-)prescription group.Autologous blood injection was used to make a mouse model of intracerebral hemorrhage.The prescription group was given traditional Chinese medicine decoction for activating blood and removing stasis and supplementing essence and marrow by gavage,and the model group and sham operation group were gavage with the same amount of normal saline respectively.Longa score was used to detect the neurological function of mice,The expression of Caspase-3,Bax,and Bcl-2 was detected by immunohistochemistry,and the protein expression of Caspase-3,Bax,and Bcl-2 was detected by Western blot.Results Compared with wild-type mice,Nrf2^(-/-)have poorer neurological function.Compared with Nrf2^(-/-)mice,the neurological function of the formula group was improved on the third day(P<0.05).The results of immunohistochemistry and Western blot showed that the expression of apoptosis genes Caspase-3 and Bax in Nrf2^(-/-)with intracerebral hemorrhage increased(P<0.05)and inhibited the anti-apoptotic factor Bcl-2(P<0.05)the expression of aggravated cell apoptosis.Conclusion The Chinese medicine decoction for activating blood and removing stasis and supplementing essence and marrow may be adjusted by Nrf2 to improve the degree of apoptosis in mice with experimental intracerebral hemorrhage.