摘要
目的探讨氯胺酮联合乙醇长期滥用对中老年小鼠海马β-淀粉样蛋白(Aβ)和Tau蛋白的影响。方法小鼠随机分为4组(n=16):空白对照组(C组)、乙醇对照组(E组)、氯胺酮注射组(K组)、氯胺酮联合乙醇灌胃组(KE组),每组分为两个亚组:3月组和6月组。小鼠饲养至12周龄开始给药,氯胺酮(30 mg/kg)和20%乙醇(2 ml/kg)分别采用腹腔注射和灌胃给药的方式,按体重给药,每天1次,连续给药3个月、6个月,空白对照组给予等量生理盐水。免疫组化法检测小鼠海马Aβ和Tau蛋白的表达,Western blotting法检测Tau蛋白在Thr231、S396和Ser404位点磷酸化程度。结果免疫组化结果:3月组小鼠海马区,K组Aβ和Tau蛋白表达量最高,随着给药时间的延长K组和KE组的Aβ及Tau蛋白表达量逐渐降低。Western blotting实验结果:6月KE组Tau蛋白与C组相比在Thr231、S396和Ser404位点磷酸化程度均有升高,其中S396位点磷酸化程度最高(增加463.1%)。结论在小鼠成年时期,氯胺酮长期滥用会导致海马区Aβ蛋白和Tau蛋白表达量增加;但在小鼠老年时期,随着氯胺酮滥用时间的延长海马Aβ和Tau蛋白表达量逐渐降低。氯胺酮乙醇合用促进了小鼠老年时期神经纤维缠结的生成。乙醇在氯胺酮导致Aβ和Tau蛋白表达过程中起到了抑制作用,可能是因为二者存在竞争性拮抗作用,也可能是乙醇摄入量的原因,其中涉及的确切机制需要进一步去研究。
Objective To investigate the effects of long-term abuse of ketamine combined with ethanol on Aβprotein and Tau protein in the hippocampus of rats.Methods The rats were randomly divided into 4 groups(n=16):control group(C),intragastric injection with ethanol(E),ketamine injection group(K),ketamine combined with ethanol intragastric injection group(KE).Each group was divied into two sub-groups:three-month group and six-month group.The rats were fed until they were 12 weeks old and given the drug.Ketamine(30 mg/kg)and 20%ethanol(2 ml/kg)were administered by intraperitoneal injection and gavage respectively every day during the study period.The expression of Aβprotein and Tau protein in the hippocampal CA3 field were explored using immunohistochemistry staining.Tau phosphorylation at the site of Thr231,S396,and Ser404 was detected using western blotting.Results Immunohistochemical results:in the threemonth group,the highest expression levels of Aβprotein and Tau protein in the hippocampus were observed in the only ketamine administered mice model.the decreased levels of Aβprotein and Tau protein in the hippocampus of the K and the KE groups weere detected with the prolongation of the administration time.Western Blot results:compared with the control group,significant increases in the phosphorylation level of Tau protein in the KE group at the site of Thr231,S396,and Ser404 were detected after drug administration for six months,with the highest phosphorylation at the site of S396(an increase of 463.1%).Conclusion Longterm abuse of ketamine increase expressions of Aβprotein and Tau protein in the hippocampus of middle-aged mice,while the expressions of Aβprotein and Tau protein in the hippocampus of elderly mice gradually decreases with the prolonged abuse of ketamine.Co-administration of ketamine and ethanol promoted the accumulation of neurofibrillary tangles(NFTs).Ethanol plays an inhibitory role in the collection of Aβprotein and Tau protein caused by ketamine,which might be due to the competitive antagonism
作者
钟磊磊
王一航
朱婷娜
蒋鸿雁
洪仕君
李慧方洁
Zhong Leilei;Wang Yihang;Zhu Tingna;Jiang Hongyan;Hong Shijun;Li Huifangjie(Kunming Medical University School of Forensic Medicine,Kunming 650500;Kunming Medical University School of College of pharmacy,Kunming 650500;Kunming Medical University Dept,of Human Anatomy and Histology&Embryology,Kunming 650500)
出处
《中国法医学杂志》
CSCD
2020年第5期490-494,共5页
Chinese Journal of Forensic Medicine
基金
国家自然科学基金资助项目(81460213)。
关键词
氯胺酮
乙醇
AΒ蛋白
TAU蛋白
中老年小鼠
Ketamine
Ethanol
Aβprotein
Tau protein
Middle-aged and elderly mice
