川蛭通络胶囊通过抑制神经炎症和氧化应激治疗小鼠脑缺血再灌注损伤

Chuanzhi Tongluo Capsules antagonizes cerebral ischemia-reperfusion injury in mice by inhibiting neuroinflammation and oxidative stress

川蛭通络胶囊(CZTL)具有活血化瘀、益气通络的功效,用于中风病中经络(脑梗死)恢复期血瘀气虚证,并在临床中得到广泛应用,但其现代药理学作用机制仍不清晰。该研究通过线栓法建立小鼠大脑中动脉缺血再灌注(MCAO/R)模型,探讨CZTL治疗缺血性脑中风的药效和作用机制。小鼠按体质量随机分为假手术组、模型组和CZTL组,CZTL组连续3 d按0.3 g·kg^(-1)·d^(-1)灌胃CZTL,最后1次灌胃1 h后,模型组和CZTL组行MCAO/R手术。再灌注24 h后,检测CZTL对小鼠神经学评分、脑梗死面积、脑水肿及脑组织病理学,同时检测脑组织匀浆活性氧自由基(ROS)、丙二醛(MDA)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平和超氧化物歧化酶(SOD)活性;利用Western blot技术检测脑组织B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Toll样受体4(TLR 4)以及磷酸化核因子-κB亚基P65(p-NF-κB P65)的表达。结果显示,CZTL显著降低MCAO/R小鼠神经学评分、脑水肿以及脑梗死面积,改善脑组织病理学,抑制脑组织ROS、MDA、IL-6、IL-1β以及TNF-α的分泌,提高SOD的活性,抑制脑组织促凋亡蛋白Bax的表达,促进抑凋亡蛋白Bcl-2的表达,并同时抑制炎症相关蛋白TLR4的表达和NF-κB P65的磷酸化。以上结果表明,CZTL对MCAO/R小鼠具有显著的保护作用,且这种保护作用与CZTL抑制神经炎症、抵抗脑组织氧化应激密切相关。

Chuanzhi Tongluo Capsules(CZTL)is effective in activating blood,resolving stasis,replenishing Qi,and dredging collaterals,which has been widely used in clinical treatment of stroke(cerebral infarction)differentiated into the syndrome of wind striking meridian and collateral in the recovery stage characterized by blood stasis and Qi deficiency.However,its modern pharmacological mechanisms of action remain unclear.This study duplicated the middle cerebral artery occlusion and reperfusion(MCAO/R)model in mice using the suture-occluded method to explore the protective effect and mechanism of CZTL on ischemic stroke.The mice were divided into the sham-operation group,model group,and CZTL group.The ones in the CZTL group were gavaged with 0.3 g·kg^(-1)·d^(-1) CZTL for three successive days.One hour after the last intragastric administration,those in the model and CZTL groups were subjected to MCAO/R.After 24 h reperfusion,the effects of CZTL on neurological deficit score,cerebral infarction area,brain edema,and brain histopathology were evaluated.The levels of reactive oxygen species(ROS),malondialdehyde(MDA),interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)and the activity of superoxide dismutase(SOD)in brain tissue homogenate were detected using corresponding assay kits.The expression of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),Toll like receptor 4(TLR4),and phosphorylated nuclear factor-κB P65 subunit(p-NF-κB P65)were assayed by Western blot.The results indicated that CZTL significantly reduced the neurological deficit score,brain edema,and infarct volume,improved the brain histopathology,inhibited the expression of ROS,MDA,IL-6,IL-1β,and TNF-αin the brain tissue,and up-regulated the activity of SOD,down-regulated the expression of pro-apoptotic protein Bax,promoted the expression of anti-apoptotic protein Bcl-2,and suppressed the expression of TLR4 and p-NF-κB P65 phosphorylation of MCAO/R mice.All these have demonstrated that CZTL has a significant protective effect against MCAO/R injury in mice,which may be related to its resistance to neuroinflammation and oxidative stress.