Hypidone hydrochloride(YL-0919)ameliorates functional deficits after traumatic brain injury in mice by activating the sigma-1 receptor for antioxidation

Traumatic brain injury involves complex pathophysiological mechanisms,among which oxidative stress significantly contributes to the occurrence of secondary injury.In this study,we evaluated hypidone hydrochloride(YL-0919),a self-developed antidepressant with selective sigma-1 receptor agonist properties,and its associated mechanisms and targets in traumatic brain injury.Behavioral experiments to assess functional deficits were followed by assessment of neuronal damage through histological analyses and examination of blood-brain barrier permeability and brain edema.Next,we investigated the antioxidative effects of YL-0919 by assessing the levels of traditional markers of oxidative stress in vivo in mice and in vitro in HT22 cells.Finally,the targeted action of YL-0919 was verified by employing a sigma-1 receptor antagonist(BD-1047).Our findings demonstrated that YL-0919 markedly improved deficits in motor function and spatial cognition on day 3 post traumatic brain injury,while also decreasing neuronal mortality and reversing blood-brain barrier disruption and brain edema.Furthermore,YL-0919 effectively combated oxidative stress both in vivo and in vitro.The protective effects of YL-0919 were partially inhibited by BD-1047.These results indicated that YL-0919 relieved impairments in motor and spatial cognition by restraining oxidative stress,a neuroprotective effect that was partially reversed by the sigma-1 receptor antagonist BD-1047.YL-0919 may have potential as a new treatment for traumatic brain injury.

S1-2 The 5-HT6 Receptor-Related Mechanism for the Cognition-Enhancing Properties of Hypidone Hydrochloride(YL-0919),A Novel Protential Antidepressant

Hypidone hydrochloride(YL-0919),the 5-HT1A/6 agonists and 5-HT reuptake inhibitor,is a novel potent antidepressant with original chemical structure.Previous studies confirmed that YL-0919 has significant antidepressant-and anxiolytic-like effects.Compared with first-line antidepressants,YL-0919 possesses rapid-onset and cognition-enhancing advantages without causing sexual disorders.Recently,it has been found that it has high affinity with 5-HT6 receptor.Objective:To study the target characteristics of YL-0919 to 5-HT6 receptors,and to explore the relationship between the 5-HT6 receptor and the cognition-enhancing,antidepressant/anxiolytic-like effects of YL-0919 and targeting mechanisms.Methods:The radioligand binding inhibition test and[35S]-GTPγS binding assay were used to evaluate the binding affinity of YL-0919 to 5-HT6 receptor in rat striatum,transient CHO cell line and stable Hela cell lines.Novel object recognition(NOR),Morris water maze(MWM)and step-down test(SD)were used to evaluate the cognition-enhancing activity of YL-0919,and the selective 5-HT6 receptor antagonist SB271046 was used to evaluate the relationship between behavioral improvement caused by YL-0919 and 5-HT6 receptor activation.To study the 5-HT6 receptor related mechanisms of YL-0919,the competitive immunofluorescence assay were used to examine the cAMP level in h5-HT6 receptor-expressed in the Hela cells Results:①Radioligand competitive binding experiments showed that YL-0919 had high binding affinity with 5-HT6 receptors in the rat striatum,the CHO cells transiently expressed the h5-HT6 receptor and the Hela cells stably expressed the h5-HT6 receptor,with Ki of 10.72,14.76 and 28.12 nM respectively;[35S]-GTPγS showed full agonist characteristics of YL-0919 in striatum and cells,with EC50 of 71.23,64.73 and 52.92 nM respectively,and the maximum efficiency(Emax)reached 100%which is the same to the 5-HT6 receptor agonist WAY208466,suggesting that YL-0919 is a full 5-HT6 receptor agonist.②Cognitive-related behavioral tests showed that subchronic oral administration of YL-0919(1.25~2.5 mg·kg-1)could significantly increase the recognition index in NOR,the entries and duration in the target quadrant,the entries crossing the platform in WMW,shortened the first time crossing the platform in MWM and the step-down latency in SD,suggesting the cognitionenhancing effects of YL-0919;compared with Vilazodone,the partial agonist of 5-HT1A receptor and 5-HT reuptake inhibitor,which of no such functions;Further study showed that 5-HT6 receptor antagonist SB271046(10 mg·kg-1)completely blocked the cognition-enhancing effects of YL-0919 without affecting the cognitive activity itself,suggesting that 5-HT6 receptor activation might be its underlying mechanisms;③Mechanism study found that YL-0919 could significantly increase cAMP levels in the Hela cells stably-expressed the h5-HT6 receptor,which could be dose-dependent blocked by SB271046.Conclusion:YL-0919 is a full agonist of 5-HT6 receptor.YL-0919 showed significant cognition-enhancing effects in various kinds of animal models,and its underlying important mechanism might be activating 5-HT6 receptor.In addition,enhancing downstream cAMP-CREB signaling pathway of 5-HT6 receptor might at least partially mediate the above process.Moreover,5-HT6 receptor activation might also be one of the mechanisms of antidepressant-and anxiolytic-like effects of YL-0919.In conclusion,this study confirmed the 5-HT6 receptor-related mechanisms of YL-0919,the 1.1 types of antidepressants,laying the experimental foundation for developing novel antidepressants with cognition-enhancing effects.

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

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。