盐酸羟哌吡酮(YL-0919)通过抑制铁死亡改善大鼠缺血性脑卒中

Hypidone hydrochloride(YL-0919)improves ischemic stroke in rats by inhibiting ferroptosis

目的 研究盐酸羟哌吡酮(代号:YL-0919)对缺血性脑卒中(IS)模型大鼠运动功能的改善作用及铁死亡相关机制。方法 选用成年雄性SD大鼠,通过大脑中动脉闭塞(MCAO)手术构建IS模型。实验随机分为假手术组、MCAO组、MCAO+YL-0919(5 mg·kg^(-1))组和MCAO+YL-0919(5 mg·kg^(-1))+铁死亡诱导剂埃拉斯汀(Era,15 mg·kg^(-1))组。给药组造模6 h后首次ip给药,此后每天给药1次。连续给药7~10 d后,通过神经功能评分、粘附物移除实验、平衡木实验、旋转棒实验、旷场实验评价YL-0919对IS后运动功能的影响;连续给药7 d后,通过TTC染色检测脑梗死面积,比色法检测大脑皮质半暗带组织中谷胱甘肽(GSH)、丙二醛(MDA)和亚铁离子(Fe^(2+))的含量,Western印迹法检测大脑皮质半暗带中谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(xCT)、酰基辅酶A合成酶长链家族成员4(ACSL4)和转铁蛋白受体1(TFR1)蛋白表达水平。结果 与假手术组相比,MCAO组大鼠神经功能评分显著升高(P<0.01),移除时间和首次接触右前爪时间明显延长(P<0.01),通过平衡木时间明显延长(P<0.01),在旋转棒上停留时间显著缩短(P<0.01),在旷场中运动距离明显缩短(P<0.01);脑梗死面积显著增大(P<0.01);皮质半暗带组织中GSH含量显著降低(P<0.01),而MDA和Fe^(2+)含量显著上升(P<0.01),GPX4和xCT蛋白表达水平显著降低(P<0.05),ACSL4和TFR1蛋白表达水平明显升高(P<0.05)。与MCAO组相比,给予YL-0919后上述变化显著逆转,而同时给予Era和YL-0919时,YL-0919的逆转作用被明显削弱。结论 YL-0919能改善大鼠IS后的运动功能损伤、减少脑梗死面积,其机制可能与抑制大脑皮质细胞铁死亡有关。

OBJECTIVE To study the way in which hypidone hydrochloride(code:YL-0919)improves motor function after ischemic stroke(IS)and explore the related mechanism.METHODS Adult male SD rats were used to establish a middle cerebral artery occlusion(MCAO)model that simulated acute IS.All animals were randomly divided into four groups:sham group,MCAO group,MCAO+YL-0919 group,and MCAO+YL-0919+erastin(Era,ferroptosis inducer)group.The drug administration groups received the first ip injection 6 h after operation,followed by continuous ip injection once per day.After 7-10 d of drug administration,the effect of YL-0919 on motor function after IS were evaluated via neu-rological function test,adhesive-removal test,rotarod test,balance beam test and open field test.After 7 d of drug administration,TTC staining was used to detect the cerebral infarction area while the colo-rimetry method was used to measure the contents of glutathione(GSH),malondialdehyde(MDA),and ferrous ions(Fe^(2+))in the penumbra of the cerebral cortex.Western blotting was used to detect the expression levels of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(xCT),acyl-CoA synthetase long-chain family member 4(ACSL4),and transferrin receptor 1(TFR1)in the cortical penumbra.RESULTS Compared with the sham group,the MCAO group showed higher neurological function scores(P<0.01),with notably prolonged time for tape removal and first contact with the right forepaw(P<0.01),spent significantly more time crossing the balance beam(P<0.01)but endured a notably shorter duration on the rotarod(P<0.01),reduced the movement distance in the open field(P<0.01),had a remarkably increased infarct area(P<0.01)but significantly level of GSH in the cortical penumbra region decreased(P<0.01),while MDA and Fe^(2+)levels were markedly increased(P<0.01).Protein expression levels of GPX4 and xCT were reduced(P<0.05),while those of ACSL4 and TFR1 were elevated(P<0.05).Compared with the MCAO group,these changes were significantly reversed after YL-0919 administration.However,when Era and YL-0919 were administered simultaneously,the reversal effect of YL-0919 was significantly weakened.CONCLUSION YL-0919 can improve motor function impairment and reduce cerebral infarction areas in rats after IS,and the mechanism may be related to the inhibition of ferroptosis.