温胆汤对精神分裂症模型大鼠海马组织PI3K,Akt和GSK3β的影响
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摘要
目的:研究温胆汤对精神分裂症模型鼠海马组织磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/糖原合成酶激酶3β(GSK3β)信号通路的影响。方法:将60只大鼠随机分为6组,分别为正常组、模型组、氯氮平组、温胆汤高、中、低剂量组,每组10只。正常组和模型组灌胃生理盐水,氯氮平组灌胃氯氮平原药20 mg·kg~(-1);温胆汤高、中、低剂量组分别灌胃温胆汤生药40,20,10 g·kg~(-1); 1次/d,共21 d。在最后1次给药2 h后,除正常组外,其余各组采用一次性左侧腹腔注射MK8010. 6 mg·kg~(-1),以诱发精神分裂症。观察并记录大鼠的刻板行为,3 d后,处死并取其海马组织待测。依照Sams Dodd和Hoffman标准对大鼠的刻板行为评分;蛋白免疫印迹法(Western blot)测定海马组织PI3K,Akt和GSK3β蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测海马组织PI3K,Akt,GSK3βmRNA表达。结果:与正常组比较,模型组大鼠刻板行为评分显著升高(P <0. 01),海马组织PI3K,Akt,GSK3β蛋白和mRNA表达水平显著下降(P <0. 01)。与模型组比较,温胆汤给药组能明显降低大鼠刻板行为评分(P <0. 05),升高海马组织PI3K,Akt,GSK3β蛋白和mRNA表达水平(P <0. 05,P <0. 01)。结论:温胆汤通过改善精神分裂症模型鼠刻板行为、升高其海马组织PI3K,Akt,GSK3β的表达,调控PI3K/Akt/GSK3信号通路,从而达到治疗精神分裂症的目的。
Objective: To research the effect of Wendantang on phosphatidylinositol-3-kinases(PI3 K)/protein kinase B(Akt)/glycogen synthase kinase 3β(GSK3β) signaling pathway of hippocampus in rats with schizophrenia. Method: Sixty SD rats were randomly divided six groups: normal group(A),model group(B),Clozapine group(C),high-dosage group Wendantang(D),medium-dosage Wendantang group(E) and low-dosage Wendantang group(F),with 10 rats in each group. The rats of normal group and model group were given normal saline. The rats of Clozapine group were given 20 mg·kg~(-1) Clozapine. The rats of high-dosage,medium-dosage,low-dosage Wendantang groups were respectively given 40,20,10 g·kg~(-1) Wendantang,once a day,for 21 days. Two hours later after the last administration with Wendantang or saline,except for group A,groups B,C,D,E,F were intraperitoneally given MK-801 0. 6 mg·kg~(-1) to establish the rat schizophrenia model. After modeling,the changes in stereotyped behavior of each group were observed and recorded. After three days,the rats were put to death,and the hippocampus tissue were tested. According to Sams Dodd and Hoffman's standard,the stereotyped behavior of rats was scored. The protein expressions of PI3 K,Akt and GSK3β of hippocampus were determined by Western blot. The mRNA expressions of PI3 K,Akt and GSK3β of hippocampus were tested by Real-time PCR. Result: Compared with A,the stereotyped behavior score of the B was significantly increased(P < 0. 01). The expressions of PI3 K, Akt, GSK3β protein and mRNA of hippocampus were significantly decreased(P < 0. 01) in rats of the B. Compared with the B,Wendantang administration groups could reduce the stereotyped behavior score(P < 0. 05),increased significantly the expressions of PI3 K,Akt,GSK3βprotein and PI3 K,Akt and GSK3β mRNA of hippocampus(P < 0. 05,P < 0. 01). Conclusion: Wendantang could regulate the PI3 K/Akt/GSK3 signaling pathway by improving the stereotyped behavior and increasing the expressions of PI3 K,Akt,GSK3β of hippocampus in rats with schizophrenia,so as to achieve the purpose of treating schizophrenia.
Objective: To research the effect of Wendantang on phosphatidylinositol-3-kinases(PI3 K)/protein kinase B(Akt)/glycogen synthase kinase 3β(GSK3β) signaling pathway of hippocampus in rats with schizophrenia. Method: Sixty SD rats were randomly divided six groups: normal group(A),model group(B),Clozapine group(C),high-dosage group Wendantang(D),medium-dosage Wendantang group(E) and low-dosage Wendantang group(F),with 10 rats in each group. The rats of normal group and model group were given normal saline. The rats of Clozapine group were given 20 mg·kg~(-1) Clozapine. The rats of high-dosage,medium-dosage,low-dosage Wendantang groups were respectively given 40,20,10 g·kg~(-1) Wendantang,once a day,for 21 days. Two hours later after the last administration with Wendantang or saline,except for group A,groups B,C,D,E,F were intraperitoneally given MK-801 0. 6 mg·kg~(-1) to establish the rat schizophrenia model. After modeling,the changes in stereotyped behavior of each group were observed and recorded. After three days,the rats were put to death,and the hippocampus tissue were tested. According to Sams Dodd and Hoffman's standard,the stereotyped behavior of rats was scored. The protein expressions of PI3 K,Akt and GSK3β of hippocampus were determined by Western blot. The mRNA expressions of PI3 K,Akt and GSK3β of hippocampus were tested by Real-time PCR. Result: Compared with A,the stereotyped behavior score of the B was significantly increased(P < 0. 01). The expressions of PI3 K, Akt, GSK3β protein and mRNA of hippocampus were significantly decreased(P < 0. 01) in rats of the B. Compared with the B,Wendantang administration groups could reduce the stereotyped behavior score(P < 0. 05),increased significantly the expressions of PI3 K,Akt,GSK3βprotein and PI3 K,Akt and GSK3β mRNA of hippocampus(P < 0. 05,P < 0. 01). Conclusion: Wendantang could regulate the PI3 K/Akt/GSK3 signaling pathway by improving the stereotyped behavior and increasing the expressions of PI3 K,Akt,GSK3β of hippocampus in rats with schizophrenia,so as to achieve the purpose of treating schizophrenia.
引文
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[19] Emamian E S. AKT/GSK3 signaling pathway and schizophrenia[J]. Front Mol Neurosci, 2012,5(33):33.
[20] Beurel E,Grieco S F,Jope R S. Glycogen synthase kinase-3(GSK3):regulation,actions,and diseases[J]. Pharmacol Ther,2015,148:114-131.
[21]田龙夫,张琦,王波涛,等.血根碱通过调控PI3K/Akt信号通路诱导胰腺癌细胞凋亡的机制[J].中国实验方剂学杂志,2018,24(19):166-171.
[22]徐厚谦,颜春鲁,张永花,等.当归补血汤通过PI3K/Akt通路对AngⅡ诱导肥大心肌细胞的保护作用[J].中国实验方剂学杂志,2018,24(2):135-139.
[23] Saijilafu,Hur E M,LIU C M,et al. PI3K-GSK3signaling regulates mammalian axon regeneration by inducing the expression of Smad1[J]. Nat Commun,2013,doi:10. 1038/ncomms3690.
[24]徐少群,徐乐,李向宇,等.电针对异氟醚诱导阿尔茨海默样病小鼠海马区PI3K/Akt信号通路的影响[J].吉林中医药,2016,36(3):294-297.
[25] ZHENG W,WANG H,ZENG Z,et al. The possible role of the Akt signaling pathway in schizophrenia[J]. Brain Research,2012,1470(35):145-158.
[26] Thiselton D L,Vladimirov V I,Kuo P H,et al. AKT1is associated with schizophrenia across multiple symptom dimensions in the Irish study of high density schizophrenia families[J]. Biol Psychiatry,2008,63(5):449-457.
[27] Amar S, Shaltiel G, Mann L, et al. Possible involvement of post-dopamine D2 receptor signalling components in the pathophysiology of schizophrenia[J].Int J Neuropsychopharmacol,2008,11(2):197-205.
[28] Emamian E S,Hall D,Birnbaum M J,et al. Convergent evidence for impaired AKTI-GSK3βsignaling in schizophrenia[J]. Nat Genet,2004,36(2):131-137.
[29] Benedetti F,Poletti S,Radaelli D,et al. Temporal lobe grey matter volume in schizophrenia is associated with a genetic polymorphism influencing glycogen synthase kinase 3-βactivity[J]. Genes Brain Behav,2010,9(4):365-371.
[2] Howes O,McCutcheon R,Stone J. Glutamate and dopamine in schizophrenia:An update for the 21st century[J]. J Psychopharmacol,2015,29(2):97-115.
[3] Karam C S,Ballon J S,Bivens N M,et al. Signaling pathways in schizophrenia:emerging targets and therapeutic strategies[J]. Trends Pharmacol Sci,2010,31(8):381-390.
[4] Singh K K. An emerging role for Wnt and GSK3signaling pathways in schizophrenia[J]. Clin Genet,2013,83(6):511-517.
[5]唐倩倩,郭小云,刘迪,等. GSK-3β参与神经精神疾病的研究进展[J].中国药理学通报,2014,30(9):1193-1196.
[6]李冀,连建伟.方剂学[M]. 4版.北京:中国中医药出版社,2016:270.
[7]万红娇,杨翠萍,丁舸.中医药治疗精神分裂症的临床研究概况[J].江西中医学院学报,2009,21(5):92-95.
[8]郭艳梅,史彦彦,李玉欣,等.中药治疗精神分裂症的研究进展[J].现代中西医结合杂志,2010,19(27):3543-3545.
[9]徐义勇,田真真,易惺钱,等.温胆汤联合抗精神病药物治疗精神分裂症疗效与安全性的Meta分析[J].中华中医药学刊,2017,35(10):2536-2540.
[10]周叔平.论温胆汤之功效与主治[J].中国中药杂志,2003,28(3):97-98.
[11]付艳丽,万红娇,朱金华,等.温胆汤对地卓西平马来酸盐诱发精神分裂症模型大鼠脑组织中Cx43 mRNA表达的影响[J].中国实验方剂学杂志,2012,18(20):173-175.
[12]朱金华,田真真,万红娇,等.温胆汤对精神分裂症模型鼠血清SOD,MDA,NO,PKC的影响[J].中国实验方剂学杂志,2013,19(13):254-257.
[13]朱金华,孙昊鑫,魏妍妍,等.温胆汤对精神分裂症模型鼠海马组织Cx43、谷氨酸及神经胶质细胞超微结构的影响[J].中药药理与临床,2014,30(5):1-5.
[14]朱金华,孙昊鑫,熊秋迎,等.温胆汤对精神分裂症模型鼠血清TNF-α,IL-6及海马组织Glu活性表达的影响[J].中国实验方剂学杂志,2014,20(14):160-164.
[15]熊怀亮,朱金华,戎文娟,等.温胆汤含药血清对谷氨酸条件下星形胶质细Cx43和GJIC功能的影响[J].中药药理与临床,2016,32(1):1-5.
[16]朱金华,田真真,戎文娟,等.温胆汤对精神分裂症大鼠海马组织NRG1、Erb B4 mRNA表达及其行为学的影响[J].中药药理与临床,2017,33(3):2-5.
[17]魏妍妍,刘丹丹,戎文娟,等.温胆汤对精神分裂症模型鼠海马PKC,p38MAPK及P-Cx43的影响[J].中国实验方剂学杂志,2015,21(11):103-106.
[18] Koros E,Rosenbrock H,Birk G,et al. The selective m Glu5 receptor antagonist MTEP, similar to NMDA receptor antagonists,induces social isolation in rats[J].Neuropsychopharmacology,2007,32(3):562-576.
[19] Emamian E S. AKT/GSK3 signaling pathway and schizophrenia[J]. Front Mol Neurosci, 2012,5(33):33.
[20] Beurel E,Grieco S F,Jope R S. Glycogen synthase kinase-3(GSK3):regulation,actions,and diseases[J]. Pharmacol Ther,2015,148:114-131.
[21]田龙夫,张琦,王波涛,等.血根碱通过调控PI3K/Akt信号通路诱导胰腺癌细胞凋亡的机制[J].中国实验方剂学杂志,2018,24(19):166-171.
[22]徐厚谦,颜春鲁,张永花,等.当归补血汤通过PI3K/Akt通路对AngⅡ诱导肥大心肌细胞的保护作用[J].中国实验方剂学杂志,2018,24(2):135-139.
[23] Saijilafu,Hur E M,LIU C M,et al. PI3K-GSK3signaling regulates mammalian axon regeneration by inducing the expression of Smad1[J]. Nat Commun,2013,doi:10. 1038/ncomms3690.
[24]徐少群,徐乐,李向宇,等.电针对异氟醚诱导阿尔茨海默样病小鼠海马区PI3K/Akt信号通路的影响[J].吉林中医药,2016,36(3):294-297.
[25] ZHENG W,WANG H,ZENG Z,et al. The possible role of the Akt signaling pathway in schizophrenia[J]. Brain Research,2012,1470(35):145-158.
[26] Thiselton D L,Vladimirov V I,Kuo P H,et al. AKT1is associated with schizophrenia across multiple symptom dimensions in the Irish study of high density schizophrenia families[J]. Biol Psychiatry,2008,63(5):449-457.
[27] Amar S, Shaltiel G, Mann L, et al. Possible involvement of post-dopamine D2 receptor signalling components in the pathophysiology of schizophrenia[J].Int J Neuropsychopharmacol,2008,11(2):197-205.
[28] Emamian E S,Hall D,Birnbaum M J,et al. Convergent evidence for impaired AKTI-GSK3βsignaling in schizophrenia[J]. Nat Genet,2004,36(2):131-137.
[29] Benedetti F,Poletti S,Radaelli D,et al. Temporal lobe grey matter volume in schizophrenia is associated with a genetic polymorphism influencing glycogen synthase kinase 3-βactivity[J]. Genes Brain Behav,2010,9(4):365-371.