[1]郭鹏 任凯夕 沈金金.木犀草素对实验性自身免疫性脑脊髓炎大鼠SPH/SPHK1/S1P1通路及神经元凋亡的影响[J].卒中与神经疾病杂志,2021,28(02):137-143.[doi:10.3969/j.issn.1007-0478.2021.02.001]
　Guo Peng,Ren Kaixi,Shen Jinjin..The effects of Luteolin on SPH/SPHK1/S1P1 pathway and neuron apoptosis in rats with experimental autoimmune encephalomyelitis[J].Stroke and Nervous Diseases,2021,28(02):137-143.[doi:10.3969/j.issn.1007-0478.2021.02.001]
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木犀草素对实验性自身免疫性脑脊髓炎大鼠SPH/SPHK1/S1P1通路及神经元凋亡的影响()
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《卒中与神经疾病》杂志[ISSN:1007-0478/CN:42-1402/R]

卷:
第28卷
期数:
2021年02期
页码:
137-143
栏目:
论 著
出版日期:
2021-04-20

文章信息/Info

Title:
The effects of Luteolin on SPH/SPHK1/S1P1 pathway and neuron apoptosis in rats with experimental autoimmune encephalomyelitis
文章编号:
1007-0478(2021)02-0137-07
作者:
郭鹏 任凯夕 沈金金
710038 西安,空军军医大学唐都医院神经内科[郭鹏 任凯夕 沈金金(通信作者)]
Author(s):
Guo Peng;  Ren Kaixi;  Shen Jinjin.
Department of Neurology, Tangdu Hospital, Air Force Military Medical University, Xi’an 710038
关键词:
木犀草素 自身免疫性脑脊髓炎 神经酰胺-鞘氨醇/鞘氨醇激酶/1-磷酸鞘氨醇受体1 神经元凋亡
Keywords:
Luteolin Experimental autoimmune encephalomyelitis Ceramide-sphingosine/sphingosine kinase/sphingosine-1-phosphate receptor 1 Neurons Apoptosis
分类号:
R742
DOI:
10.3969/j.issn.1007-0478.2021.02.001
文献标志码:
A
摘要:
目的 探讨木犀草素(Luteolin)对实验性自身免疫性脑脊髓炎(Experimental autoimmune encephalomyelitis,EAE)大鼠神经酰胺-鞘氨醇(Sphingosine,SPH)/鞘氨醇激酶1(Sphingosine kinase 1,SPHK1)/1-磷酸鞘氨醇受体1(Sphingosine-1-phosphate 1,S1P1)通路蛋白表达水平及神经元凋亡的影响。方法 取雌性Wistar大鼠,采用注射豚鼠脊髓免疫抗原法建立EAE模型,将造模成功的50只大鼠随机分为模型(EAE)组,Luteolin低(5mg/kg)、中(25 mg/kg)、高(50 mg/kg)剂量组,芬戈莫德(FTY720)阳性对照组(61.7 mg/kg),每组各10只; 另取10只雌性Wistar大鼠,注射等量生理盐水,作为空白对照(Control)组; 各组于造模15 d后开始给药,Luteolin低、中、高剂量组经腹腔注射相应剂量Luteolin药物,芬戈莫德(FTY720)阳性对照组灌胃给予相应剂量的FTY720,EAE组和Control组灌胃和腹腔注射等量生理盐水,各组连续给药14 d,2次/d; 各组大鼠于末次给药12 h后对大鼠EAE临床症状进行评分; 取脊髓组织,用苏木精-伊红染色(Hematoxylin eosin,HE)及固蓝(Luxol fast blue,LFB)染色法检测大鼠髓鞘组织病理表现及脱髓鞘面积; 原位缺口末端转移酶标记法(TdT-mediated dUTP nick and labeling,TUNEL)法检测髓鞘组织中神经元的凋亡情况并计算凋亡率; 以蛋白免疫印迹法(Western Blot)检测髓鞘组织中通路蛋白SPHK,SPH,S1P1及凋亡蛋白半胱氨酸天冬氨酸蛋白酶-12(Cystein-asparate protease-12,Caspase-12)蛋白相对表达水平。结果 与Control组比较,EAE组大鼠EAE临床症状评分、脊髓组织炎性细胞浸润和髓鞘脱失程度、神经元凋亡率、髓鞘组织中SPH,SPHK1,S1P1及Caspase-12蛋白表达水平均升高(P<0.05); 与EAE组比较,Luteolin低、中、高剂量组及芬戈莫德(FTY720)阳性对照组大鼠EAE临床症状评分、脊髓组织炎性细胞浸润和髓鞘脱失程度、神经元凋亡率、髓鞘组织中SPH,SPHK1,S1P1及Caspase-12蛋白表达水平均降低(P<0.05),且Luteolin各剂量组上述指标水平呈剂量依赖性; Luteolin高剂量组与芬戈莫德(FTY720)阳性对照组的上述指标水平均无明显差异(P>0.05)。结论 Luteolin可能通过抑制EAE模型大鼠髓鞘组织SPHK,SPH,S1P1蛋白表达来抑制EAE大鼠脱髓鞘病变及神经元凋亡。
Abstract:
ObjectiveTo investigate the effects of Luteolin on the ceramide-sphingosine(SPH)/sphingosine kinase(SPHK1)/sphingosine-1-phosphate receptor 1(S1P1)pathway protein expression levels and neuron apoptosis in rats with experimental autoimmune encephalomyelitis(EAE).Methods The female Wistar rats were selected, EAE model was established by injecting spinal cord immune antigen of guinea pigs, and 50 rats were randomly divided into model(EAE)group, Luteolin low(5 mg/kg), medium(25 mg/kg)and high(50 mg/kg)dose groups, and fingorod(FTY720)positive control group(61.7 mg/kg), with 10 rats in each group. In addition, another 10 female Wistar rats were injected with the same amount of normal saline as the control group. They were given drugs after 15 days of model in each group, corresponding dose of Luteolin was injected intraperitoneally in low, middle and high dose groups in the positive control group, FTY720 was given by gavage, and EAE group and control group were given the same amount of saline by gavage and intraperitoneal injection, continuous administration for 14 days, twice a day. The clinical symptoms of EAE were evaluated after 12 hours of the last administration, the spinal cord was taken, and the pathological damage and demyelination area of myelin tissue in rats were measured by hematoxylin eosin(HE)and solid blue(LFB)staining, terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)was used to detect the apoptosis of neurons in myelin tissues and calculate the apoptosis rate. In addition, Western blotting was used to detect relative expression levels of SPHK, SPH, S1P1 and caspase-12 in myelin tissues.Results Compared with those in control group, the EAE clinical symptom score, inflammatory cell infiltration and demyelination, neuron apoptosis rate, protein expression levels of SPH, SPHK1, S1P1 and caspase-12 in myelin tissue of rats in EAE group were higher(P<0.05). Compared with those in EAE group, the EAE clinical symptom score, inflammatory cell infiltration and demyelination, neuron apoptosis rate, protein expression levels of SPH, SPHK1, S1P1 and caspase-12 in myelin tissue of rats in Luteolin low, medium, high dose groups and positive control group were lower(P<0.05), and the above parameters of Luteolin groups were dose-dependent, while there was no significant difference between the Luteolin high dose group and the positive control group(P>0.05)Conclusion Luteolin might inhibit demyelination and neuron apoptosis of EAE rats by inhibiting the SPHK, SPH and S1P1 protein expressions.

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更新日期/Last Update: 2021-04-20