摘要
目的探究五味子木脂素(Schisandra chinensis lignans,SCL)改善睡眠剥夺(sleep deprivation,SD)模型大鼠神经细胞凋亡和线粒体损伤的作用及机制。方法将48只大鼠分为对照(Control)组、睡眠剥夺模型(SD)组、莫达非尼(modafinil,MOD)组(阳性对照)和不同浓度SCL治疗组。Morris水迷宫和Y迷宫实验分别检测各组大鼠的逃避潜伏期和行为正确率,HE染色检测大鼠海马组织病理损伤,TUNEL染色检测细胞凋亡。JC-1染色和ELISA法分别检测各组大鼠脑组织或细胞线粒体膜电位(mitochondrial membrane potential,MMP)水平和ROS水平。ROS诱导剂2,3-二甲氧基-1,4-奈萘(2,3-dimethoxy-1,4-naphthoquinone,DMNQ)刺激HT-22细胞建立体外神经细胞损伤模型。CCK-8检测细胞活力。蛋白质印迹检测细胞中TLR4,MyD88和p-NF-κB P65蛋白表达。结果与Control组相比较,SD组大鼠的逃避潜伏期增加,行为正确率降低;接受Mod或SCL治疗的大鼠逃避潜伏期降低且行为正确率较SD组增加(P_均<0.05)。此外,与Control组比较,SD组大鼠海马组织病理损伤明显,细胞凋亡增加,MMP水平降低且ROS水平增加。Mod或SCL干预则改善了SD组大鼠的海马组织损伤和细胞凋亡,减少了神经细胞线粒体损伤(P_均<0.05)。细胞实验结果显示,与Control组比较,DMNQ组HT-22细胞活力降低、凋亡增加、线粒体损伤和ROS水平增加。而Mod或SCL处理则显著改善了DMNQ导致的HT-22细胞损伤。此外,与Control组比较,DMNQ组细胞中TLR4、MyD88和p-NF-κB P65蛋白质表达升高,Mod或SCL干预显著抑制DMNQ处理的HT-22细胞中TLR4、MyD88和p-NF-κB P65水平。结论SCL通过抑制TLR4-MyD88-NF-κB通路激活改善睡眠剥夺大鼠神经细胞凋亡和线粒体损伤。
Objective To explore the molecular mechanism by which Schisandra chinensis lignans(SCL)improve neuronal apoptosis and mitochondrial damage in the sleep deprivation(SD)model rats.Methods Forty-eight rats were divided into 6 groups:Control group,SD group,modafinil(MOD)group(positive control),and SCL treatment groups at different concentrations.The Morris water maze and Y-maze tests were used to assess the escape latency and behavioral accuracy of rats in each group,respectively.HE staining was used to detect hippocampal tissue pathological damage in rats,and TUNEL staining was used to detect cell apoptosis.JC-1 staining and ELISA were used to detect the levels of mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)in rats’brain tissues and cells.An in vitro neuronal damage model was established using the ROS inducer 2,3-dimethoxy-1,4-naphthoquinone(DMNQ)to stimulate HT-22 cells.CCK-8 was used to measure cell viability.Western blotting was used to detect the expression levels of TLR4,MyD88,and p-NF-κB P65 in cells.Results Compared with those in the Control group,the escape latency of SD rats increased,and behavioral accuracy decreased.In contrast,rats treated with Mod or SCL showed reduced escape latency and increased behavioral accuracy(all P<0.05).Additionally,compared with rats in the Control group,rats in the SD group exhibited significant hippocampal tissue pathological damage,increased cell apoptosis,decreased MMP levels,and increased ROS levels.Mod or SCL intervention improved hippocampal tissue damage and cell apoptosis and reduced neuronal mitochondrial damage in SD rats(all P<0.05).Cellular experimental results showed that HT-22 cells in the DMNQ group had reduced viability,increased apoptosis,and increased mitochondrial damage and ROS levels when compared with those in the Control group.Mod or SCL treatment significantly improved the DMNQ-induced HT-22 cell damage.Moreover,compared with those in the Control group,the protein expression levels of TLR4,MyD88,and p-NF-κB P65 in the DMNQ group cells were elevated,and Mod or SCL intervention significantly suppressed the levels of TLR4,MyD88,and p-NF-κB P65 in DMNQ-treated HT-22 cells.Conclusion SCL improves neuronal apoptosis and mitochondrial damage in SD rats by inhibiting the activation of the TLR4-MyD88-NF-κB pathway.
作者
赵艺
李华
胡霞
常海霞
ZHAO Yi;LI Hua;HU Xia;CHANG Haixia(Department of Nephropathy,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi,830011,China;Department of Cardiovascular Medicine,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi,830011,China;College of Nursing,Xinjiang Medical University,Urumqi,830011,China;Department of Nursing,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi,830011,China)
出处
《医学分子生物学杂志》
2025年第5期423-429,共7页
Journal of Medical Molecular Biology
基金
新疆维吾尔自治区自然科学基金(No.2022D01C566)。
关键词
睡眠剥夺
五味子
木脂素
Toll样受体4
线粒体损伤
sleep deprivation
Schisandra chinensis
lignans
toll-like receptor 4
mitochondrial damage
