探索CUMS诱导小鼠抑郁模型中促炎因子IL-17A的参与机制以及海参磷脂的干预作用
摘要
抑郁症的假说众说纷纭，近年来，巨噬细胞-T淋巴细胞假说认为过度激活的巨噬细胞与淋巴细胞所分泌的过多白细胞因子可以刺激下丘脑-垂体-肾上腺（HPA）轴，导致糖皮质激素（GC）过度分泌，通过促肾上腺皮质激素释放因子受体2（CRFR2）激活小胶质细胞，引起中枢炎症和活性氧的释放，造成神经元的损伤以及神经递质缺失，最终引发抑郁。已知在外周免疫炎症激活时，高浓度的白介素-6（IL-6）与低浓度的转化生长因子-β（TGF-β）共同诱导T辅助细胞17（Th17）分化，产生促炎因子白介素-17A（IL-17A），加剧外周炎症。研究证明，IL-17A可以改变血脑屏障通透性，使得在炎症发生时IL-17A和Th17浸润中枢。最新发现脑中也存在淋巴管道和淋巴细胞，且小胶质细胞表面存在IL-17A的受体，提示Th17细胞分泌的IL-17A与中枢炎症的发生相关。已知在中枢炎症发生时，小胶质细胞过度激活可以产生过量IL-6，且中枢炎症可抑制星形胶质细胞分泌TGF-β，这为Th17在中枢的分化创造了有利条件。在实验性脑脊髓炎（EAE）的发病过程中，Th17与IL-17A参与了中枢炎症，且IL-17A通过激活胶质细胞表面受体导致EAE的中枢炎症加剧。说另外一项关于银屑病（一种由IL-17引起的自身免疫病）的临床试验证明IL-17抑制剂，能够显著改善银屑病患者的抑郁症状。进一步研究表明，在抑郁症发生时，外周的Th17细胞分化程度及IL-17浓度均与患者抑郁程度呈线性相关。但是在抑郁症的中枢炎症中，Th17细胞与IL-17A的相关变化，及其与小胶质细胞，神经递质，行为变化的关系尚未研究。
由于抑郁症的发病机制较为复杂，目前的药物治疗效果差，副作用显著，因此探究抑郁症的发病机制和寻找高效低副作用的天然药物显得十分重要。海参作为天然的保健食品，具有抗肿瘤、调节免疫等功效。而海参磷脂作为海参体壁的主要成分，具有抗氧化应激、保护神经和改善记忆等功效。但是海参磷脂对抑郁症的治疗作用尚未报道。
根据以上介绍，本研究提出关键问题：在抑郁症模型的脑内，IL-6与TGF-β能否诱导Th17细胞在中枢的分化和分泌IL-17A，IL-17A在抑郁症的中枢炎症以及抑郁相关的行为和病理变化中扮演的角色是什么？海参磷脂能否通过改善Th17-Treg细胞的免疫功能，而达到抗抑郁作用？因此本实验选择慢性不可预知性应激（CUMS）模型，探究IL-17A在抑郁症的中枢炎症中的相关变化，以及海参磷脂对抑郁症的干预作用。孕妇便秘怎么办
方法：
1.本实验选择慢性不可预知性应激（CUMS）诱导模型作为研究抑郁症的对象，以海参磷脂（50、100mg/kg/day）灌胃治疗；
2.糖水偏好度、高架十字迷宫、旷场、悬尾检测慢性应激模型诱导的抑郁和焦虑样行为；
3.高效液相色谱（HPLC）检测大脑皮层和海马区神经递质和代谢物的含量；
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4.RT-PCR及ELISA检测外周血中GC的浓度、中枢炎症因子的浓度、Th17与Treg细胞转录因子以及IL-17A受体的表达、海马脑区小胶质细胞的激活；
结果：
1.应激造成小鼠糖水偏好度、高架开放臂探索次数和时间、旷场探索能力与活跃度均显著下降，高架封闭臂潜伏期、悬尾不动时间显著增加。海参磷脂（100mg/kg）有显著的改善作用；
2.应激造成小鼠大脑皮层及海马区的五羟色胺（5-HT）、多巴胺（DA）含量均显著减少，代谢物五羟吲哚乙酸（5-HIAA）、二羟基苯乙酸（DOPAC）均显著增加，且5-HT/5-HIAA比值与DA/DOPAC比值均显著降低。海参磷脂（100mg/kg）有显著改善作用；
3.应激引起小鼠外周GC过度释放，增加中枢炎症因子IL-6、IL-23、IL-17A及转录因子STAT3、RoRa和受体IL-17RA、IL-17RC的表达，海马区Iba-1表达增加，TGF-β显著降低，海参磷脂（100mg/kg）有一定的改善效果；
信息技术培训心得体会结论：
1.海参磷脂抑制了促炎因子IL-6、IL-17A、IL-23的释放，抑制了Th17细胞转录因子STAT3、RoRa的表达，降低了IL-17RC的表达，抑制了小胶质细胞Iba-1的表达，增加了TGF-β表达，改善了慢性应激
诱导的抑郁样行为和神经递质含量下降和代谢增加；
2.IL-17参与中枢炎症的原因可能是应激诱导了Th17分化，产生的过量IL-17A 激活胶质细胞表面受体IL-17RA和IL-17RC，引起小胶质细胞促炎表型的增加，加剧抑郁症的中枢炎症。
关键词：慢性应激；抑郁症；中枢炎症；IL-17A；海参磷脂
Explore the mechanism of proinflammatory factor道奇摩托
IL-17A participation in CUMS-induced depression model of mice and intervention effect of
a cucumber phospholipids
Abstract
There are many different hypothes about depression.In recent years,the Monocyte-T-Lymfocyte hypothesis believes that excessively activated macrophages and lymphocytes crete too much leukocyte factor and the immune respon can stimulate the hypothalamic-pituitary-adrenal(HPA)axis,Leading to excessive cretion of glucocorticoids(GC),activation of microglia through corticotropin-releasing factor receptor2(CRFR2),causing the relea of neuroinflammation a
nd reactive oxygen species, causing neuronal damage and loss of neurotransmitters,And eventually led to depression. It is known that when peripheral inflammation is activated,high concentrations of interleukin-6(IL-6)and low concentrations of transforming growth factor-β(TGF-β) together induce T helper cell17(Th17)differentiation to produce the proinflammatory factor interleukin-17A(IL-17A),exacerbating peripheral inflammation.Studies show that IL-17A can change the permeability of the blood-brain barrier,making IL-17A and Th17 infiltrate the central nervous sysytem(CNS)when inflammation occurs.It is newly found that there are also lymphatic vesls in the brain,and there are IL-17A receptors on the surface of microglia,suggesting that Th17cells crete IL-17A and cau central inflammation.It is known that when neuroinflammation occurs,excessive activation of microglia can produce excessive IL-6,and neuroinflammation can cau the decrea of TGF-βcreted by astrocytes,which creates favorable conditions for the differentiation of Th17in the central.In the pathogenesis of experimental autoimmune encephalomyelitis (EAE),Th17and IL-17A participated in the neuroinflammation of the dia,and IL-17A incread the neuroinflammation of EAE by activating glial cell surface receptors.It shows that Th17and IL-17A can participate in neuroinflammation of brain dia.Another clinical trial on psoriasis(an autoimmune dia caud by IL-17)proved that after treatment with IL-17inhibitors,the depressive symptoms of patients with psoriasis complicated by depression also improved,Indicating that IL-17may be involved in the o
ccurrence of depression.Studies have shown that when depression occurs,the differentiation of Th17cells and the concentration of IL-17in the periphery are positively correlated with the degree of depression.However,in the neuroinflammation of depression,
the changes of Th17cells and IL-17A,and their relationship with microglia, neurotransmitters,and behavioral changes have not been studied.二元一次方程定义
Becau the pathogenesis of depression is complicated and the side effects of drugs are significant,it is very important to explore the pathogenesis of depression and find natural drugs with high efficiency and low side effects.As a natural health food,a cucumber has the functions of anti-tumor and immune regulation,while a cucumber phospholipids as the main component of the body wall of a cucumber has the effects of anti-oxidative stress,neuroprotective effect and memory improvement.The therapeutic effect of a cucumber phospholipids on depression has not been reported.
According to the above introduction,this study rais the key question:In the brain of depression model,can IL-6and TGF-βinduce Th17cell differentiation and crete IL-17A,IL-17A in the neuroinflammation of depression,and what is the role of depression-related behaviors and pathologic
al changes?Can a cucumber phospholipids achieve antidepressant effects by improving the immune function of Th17-Treg cells? Therefore,this experiment lects the Chronic Unpredictable Mild Stress(CUMS)model to explore the related changes of IL-17A in the neuroinflammation of depression,and the intervention effect of a cucumber phospholipids on depression.大肠面线
Method:
1.In this experiment,the CUMS-induced model was lected as the object of depression rearch,which was treated by gavage with a cucumber phospholipid(50, 100mg/kg/day);
2.Depression-and anxiety-like behavior induced by chronic stress was tested by sucroce preference,elevated plus maze,“open field”,tail suspension;
3.High performance liquid chromatography(HPLC)was ud to detect the content of neurotransmitters and metabolites in the cerebral cortex and hippocampus;
4.RT-PCR and ELISA was ud to detect the concentration of GC in the peripheral blood,the concentration of neuroinflammation factors,the expression of Th17and Treg cells transcription factors and IL-17A receptors,and the activation of microglia in the hippocampus;
Result:
辞职报告范文1.Stress caud a significant decrea in mice's sucroce preference,the number and time of exploration of the open arms in elevated plus maze,the exploration ability and activity of the“open field”,and the latency of the clod arms in elevated plus maze and the totalimmobility incread significantly.Sea cucumber phospholipids(100mg/kg)has a significant improvement effect;
2.Stress caud a significant decrea in the content of rotonin(5-HT)and dopamine(DA)in the cerebral cortex and hippocampus of mice,and a significant abolites pentahydroxyindoleacetic acid(5-HIAA)and dihydroxyphenylacetic acid (DOPAC),the5-HT/5-HIAA ratio and DA/DOPAC ratio were significantly reduced.Sea cucumber phospholipids(100mg/kg)has a significant improvement effect;
3.Stress caud excessive relea of peripheral GC in mice,IL-6,IL-23,IL-17A, STAT3,RoRa,IL-17RA,IL-17RC in CNS all incread significantly,the expression of Iba-1in the hippocampus incread,TGF-βsignificantly reduced,a cucumber phospholipids(100mg/kg)has a certain improvement effect;
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Conclusion:
1.Sea cucumber phospholipids inhibit the relea of proinflammatory factors IL-6, IL-17A,IL-23,inhibit the expression of Th17cell transcription factors STAT3,RoRa, reduce the expression of IL-17RC,and inhibit the microglia Iba-1expression,increa TGF-βexpression and improve CUMS-induced depression-like behaviors and loss of neurotransmitters;
2.The cau of IL-17involvement in neuroinflammation may be that stress induces Th17differentiation,and excess IL-17A produced activates glial cell surface receptors IL-17RA and IL-17RC,causing an increa in the proinflammatory phenotype of microglia, neuroinflammation that exacerbates depression;
Keywords:CUMS;Depression;Neuroinflammation;IL-17A;Sea cucumber phospholipids
