JN:下丘脑中的白介素-6可以防止肥胖,并参与神经发生的调节。

2021-09-30 从医路漫漫 MedSci原创

在运动条件下产生的白介素-6 (IL-6)在下丘脑中起到减少肥胖相关炎症的作用,并起到对食物摄入和能量消耗的控制。在下丘脑中,IL-6的一些有益作用归因于它的神经诱导特性。

背景:在运动条件下产生的白介素-6 (IL-6)在下丘脑中起到减少肥胖相关炎症的作用,并起到对食物摄入和能量消耗的控制。在下丘脑中,IL-6的一些有益作用归因于它的神经诱导特性。然而,在下丘脑中,假定的IL-6神经源性作用尚未被探索,其平衡能量摄入的潜力可能是预防或减轻肥胖的一种方法。

方法:采用野生型(WT)和IL-6敲除型(KO)小鼠,研究IL-6诱导神经发生的能力。我们采用溴脱氧尿苷(BrdU)细胞标记、免疫荧光和实时荧光定量PCR检测神经发生标志物和神经递质的表达。我们从KO中制备了IL-6诱导的下丘脑神经前体细胞,为进一步研究IL-6的神经分化作用提供了体外模型。此外,我们分析了单细胞RNA测序数据,并测定了IL-6和IL-6受体在小鼠下丘脑特定细胞类型中的表达。

结果:IL-6在下丘脑低表达,局限于小胶质细胞和伸缩细胞,而IL-6受体表达于小胶质细胞、室管膜细胞、内皮细胞和星形胶质细胞。外源性IL6可减少饮食诱导的肥胖。在非亲缘小鼠中,肥胖抵抗伴随着下丘脑中IL6表达的增加。IL6诱导下丘脑和神经祖细胞中神经发生相关基因的表达,无论是野生型小鼠还是KO小鼠。

图1 外源性IL-6对热量摄入和体质量的影响。野生型小鼠喂饲饲料或高脂饲料(HFD),腹腔注射外源性IL6或生理盐水(如A所示),测定喂食(B,C)和高脂饮食(D,E)小鼠体质量(B,D)和热量摄入量(C,E)。在所有实验中,n=5只小鼠/组。数据分析采用重复测量的双因素方差分析和Bonferroni后验。**p<0.01;*p<0.001

图2 下丘脑IL-6的转录表达。为了鉴别肥胖倾向(H)和肥胖抵抗(L)小鼠,我们执行了A中描述的程序。用实时定量PCR(B)检测下丘脑IL6的表达。单次侧脑室注射棕榈酸酯,收集下丘脑(C)测定IL 6(D)、肿瘤坏死因子α(E)和IL 1β(F),n=7~8m i c e/g r o u p;D F,n=5 m i c e/g r o u p,*p<0.0 5,**p<0.0 0 1。0 1,*p<0。0 0 1。在b a r图中,结果以平均值±标准差表示。

图3 IL6对下丘脑细胞增殖和存活的影响。小鼠按A和D中描述的方案给予IL6和BrdU治疗。取下丘脑内侧基底,用免疫荧光和共聚焦显微镜进行分析,用20倍和40倍倍镜进行细胞计数。有代表性的图像有增殖实验(INB)和存活实验(De)。给出了细胞增殖实验(INC)和存活实验(F)的总BrdU计数。两种检测方法均为6只小鼠/组。在条形图中,结果以平均值±标准差表示

图4 IL6在下丘脑诱导神经发生。根据INA所描述的方案,小鼠接受单次腹腔注射IL6。实时荧光定量PCR检测下丘脑Sox6(B)和Sox2(C)的表达。测定肥胖倾向(H)和肥胖抵抗(L)小鼠下丘脑中Sox6(D)和Doublecortin(E)的转录表达,并用单剂量IL6和BrdU治疗,如图4D所示。取下丘脑基底内侧,用免疫荧光和共聚焦显微镜进行分析;细胞计数采用20倍和40倍放大。总BrdU/NeuN计数用G(神经发生实验)表示,B,CN=5只/组,D,EN=8-10只/组,F,G,n=5只/组,B组,CN=5只/组,D,EN=8-10只/组,F,G,n=5只/组。*p<0.05,**p<0.01。在条形图中,结果以平均值±标准差表示

图5 IL6缺乏易致肥胖。用1.0 ng外源性IL-6(A)对野生型(WT)和IL-6基因敲除(KO)小鼠血清进行IL-6检测。体重测定是在8周龄(B)时进行的。用实时定量PCR方法检测7天龄(C,E)和8周龄(D,F)小鼠下丘脑组织中神经递质(C,D)和神经发生相关基因(E,F)的表达,An=3~4m i c e/g r o u p;n=2 5只小鼠/组;C,en=5~7只小鼠/组;D,Fn=9m i c e/g r o u p。*p<0.001,*p<0.05。在条形图中,结果以平均值±标准差表示

结论:IL6可诱导WT小鼠下丘脑神经发生相关基因的表达。在KO小鼠中,IL6的神经源性作用被保留,而新的完全分化的前促黑素 (POMC)和神经肽Y (NPY)神经元出现或延迟或紊乱。

原文出处:

Bobbo VC,  Engel DF,  Jara CP,et al, Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis.J Neuroinflammation 2021 Aug 31;18(1)

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    2021-10-02 xlwang2696

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