Endotoxin Induces Interleukin-1β and Nitric Oxide Synthase mRNA in Rat Hypothalamus and Pituitary
Endotoxin Induces Interleukin-1β and Nitric Oxide Synthase mRNA in Rat Hypothalamus and Pituitary
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内毒素诱导大鼠下丘脑和垂体中的白介素-1β 和一氧化氮合酶 mRNA
DOI：
10.1159/000054305
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发表时间：
1998
影响因子：
4.1
通讯作者：
A. Grossman
中科院分区：
文献类型：
--
作者：
M. Satta;R. Jacobs;G. Kaltsas;A. Grossman
一氧化氮(NO)和一氧化碳(CO)气体可能参与了下丘脑-垂体-肾上腺轴(HPA)的调节。在大脑中，NO由两种形式的一氧化氮合酶(NOS)合成，一种是构成神经元形式(NNOS)，另一种是诱导形式(INOS)。还有一种结构性的血红素加氧酶(HO2)和一种诱导型(HO1)，它们产生一氧化碳。因此，我们研究了外周注射脂多糖对下丘脑、垂体和肝脏中这些酶的基因表达以及白细胞介素1β(IL-1β)基因表达的影响。雄性Wistar大鼠(体重200~250g)，将内毒素(大肠杆菌，055B5)溶于无菌生理盐水[250µg/kg(第一组)、2.5 mg/kg(第二组)、6.25 mg/kg(第三组)]的无菌生理盐水中，按0.5ml的最终体积给予注射，对照组仅注射生理盐水。第一组和第二组分别于内毒素后1、3、8和24 h(每组4只)，第三组在3 h后提取下丘脑、垂体和肝脏的总RNA，用标准的逆转录酶方法扩增c DNA。双重聚合酶链式反应是标准化的，目的是量化与看家基因β-肌动蛋白相关的特定基因的表达。研究的特异性基因有诱导型一氧化氮合酶、一氧化氮合酶、HO1、HO2和IL-1β。聚合酶链式反应产物在琼脂糖凝胶上分离，并对条带进行密度分析，从而进行半定量。第二组大鼠下丘脑、垂体和肝脏均有诱导型一氧化氮合酶和IL-1β的表达，3h达高峰(p<；0.001)，24 h恢复至基础水平，基础状态和内毒素诱导后肝脏均未见神经元型一氧化氮合酶的表达，下丘脑和脑垂体神经元型一氧化氮合酶在基础状态下和内毒素诱导后无明显变化。第一组大鼠肝、脑组织中均有诱导型一氧化氮合酶和IL-1β的表达，但与第二组和第三组相比均有延迟，IL-1β的表达高峰出现在8h(p<；0.05)，24 h恢复到基线水平，基础状态下三种组织均有HO1和HO2的表达，第二组肝组织中HO1在1h表达增加，第三组在3h表达。下丘脑HO1、HO2在各剂量、各时间点均无明显变化。结论：不同剂量的内毒素均可诱导大鼠下丘脑和垂体中IL-1、β和iNOS的表达。我们推测，虽然IL-1β可能介导了内毒素对HPA的刺激，但这种反应的反向调节可能不涉及任何产物。
The gases nitric oxide (NO) and carbon monoxide (CO) may be involved in hypothalamo-pituitary-adrenal axis (HPA) modulation. In the brain, NO is synthesized by two forms of NO synthase (NOS), a constitutive neuronal form (nNOS) and an inducible form (iNOS). There are also a constitutive heme oxygenase (HO2) and an inducible form (HO1) which generate CO. We have therefore investigated the effect of peripheral lipopolysaccharide (LPS) administration on the gene expression of these enzymes along with interleukin-1β (IL-1β) gene expression in the hypothalamus, pituitary and liver. Male Wistar rats (200–250 g body weight) were injected intraperitoneally with endotoxin (Escherichia coli, 055 B5) dissolved in sterile normal saline [250 µg/kg first group, 2.5 mg/kg (second group) and 6.25 mg/kg (third group)] in a final volume of 0.5 ml, or saline alone in the control group. The first and the second groups were studied 1, 3, 8 and 24 h after LPS (n = 4 per group); the third group was studied at 3 h. Total RNA was extracted from the hypothalamus, pituitary and liver, and cDNA was made using standard reverse transcriptase methods. Duplex polymerase chain reaction (PCR) was standardised in order to quantify the expression of a specific gene in relation to the ‘house-keeping’ gene β-actin. The specific genes studied were iNOS, nNOS, HO1, HO2 and IL-1β. The PCR products were separated on agarose gel and densitometric analysis of the bands allowed semi-quantification. In the second group, iNOS and IL-1β were induced in hypothalamus, pituitary and liver, showing a peak at 3 h (p < 0.001), returning to baseline levels at 24 h. Neuronal NOS was not expressed in the liver under basal conditions or after LPS; in the hypothalamus and pituitary, nNOS was expressed basally but there was no change after LPS. In the first group, iNOS and IL-1β were again induced in all three tissues studied, but with a delayed time course compared to the second and third groups; the peak change for IL-1β occurred at 8 h (p < 0.05), again returning to baseline levels at 24 h. The peak for iNOS occurred at 24 h. HO1 and HO2 were expressed in all three tissues under basal conditions; HO1 was increased at 1 h in the liver in the second group, and at 3 h in the pituitary in the third group. There was no change in either HO1 or HO2 in the hypothalamus at any dose at any time point. We conclude that IL-1β and iNOS are induced in rat hypothalamus and pituitary following various doses of endotoxin. We speculate that while IL-1β may mediate stimulation of the HPA by endotoxin, NO generation may be involved in the counter-regulation of this response.
一氧化氮可能介导卵巢类固醇诱导的黄体生成激素激增的证据：兴奋性氨基酸的参与。
影响因子：
4.8
作者：
Bonavera,JJ;Sahu,A;Kalra,PS;Kalra,SP
通讯作者：
Kalra,SP
合成生长激素的垂体前叶细胞上 p80 I 型和 p68 II 型白细胞介素-I 受体的双重表达。
DOI：
10.1210/endo.137.9.8756580
发表时间：
1996
期刊：
Endocrinology.
影响因子：
--
作者：
French,RA;Zachary,JF;Dantzer,R;Frawley,LS;Chizzonite,R;Parnet,P;Kelley,KW
通讯作者：
Kelley,KW