Molecular biological studies on the roles of NFkappaBETA and NF-IL6 in the experimental glomerulonephritis and diabetic nephropathy.

NFkappaBETA和NF-IL6在实验性肾小球肾炎和糖尿病肾病中作用的分子生物学研究。

基本信息

批准号：
08457289
负责人：
HAYASHI Matsuhiko
金额：
$ 2.18万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457289/
关键词：
NFkappaBETA IkappaBETA cytokine mesangial cells RAGE NF-IL6

项目摘要

To reveal the effcnts of cytokines on the cultured mesangial cells, we examined the interactions between endothelin, IL-1beta, TNF-alpha, and lipopolysaccharide. It was shown that endothelin suppress the induction of NO synthase by these cytokines, which are known to exert their action via NFkappaBETA. We have also made a vector, which has 6 NFkappaBETA binding cite in the upstream of luciferase, and transfected it to mesangial cells. In these transfected cells, angiotensin II induced 6-fold increase in luciferase activity, suggesting that angiotensin II activates NFkappaBETA. From these results, it was suggested that NFkappaBETA plays an important role in the effects of cytokines on mesangial cells, next we made mutated IkappaBETA. This IkappaBETA lacks phosphorilation cite, which is necessary for the atctivation of NFkappaBETA. Therefore, the overexpression of mutated IkappaBETA should abolish the effects of cytokines. In the mesangial cells with the adenovirus-mediated transfection of mutated IkappaBETA, TNF-alpha showed enhanced induction of apoptosis, compared with wild type cells. It was also shown that induction of apoptosis was enhanced with the transfection of mutated IkappaBETA by IL-1 and lipopolysaccharides. From these results, it was shown that NFkappaBETA was a suppressor of apoptosis. Next, we tried to infect the adenovirus to the kidneys for the transfection of mutated IkappaBETA, although the injected adenovirus to the renal artery infected to only proximal tubules. Currently, we are developing a technique for the in vivo transfer of the mutated IkappaBETA to the kidney.

为了揭示细胞因子在培养的细胞上的EFFCNT，我们检查了内皮素，IL-1Beta，TNF-Alpha和脂多糖之间的相互作用。结果表明，内皮素抑制了这些细胞因子对NO合酶的诱导，已知通过NFKappabeta发挥作用。我们还制作了一个载体，该载体在荧光素酶上游中具有6个Nfkappabeta结合，并将其转染至膜细胞。在这些转染的细胞中，血管紧张素II诱导荧光素酶活性增加6倍，表明血管紧张素II激活NFKappabeta。从这些结果中可以提出，NFKappabeta在细胞因子对肾小球细胞的作用中起重要作用，接下来我们使Ikappabeta突变。这种iKappabeta缺乏磷的引用，这对于Nfkappabeta的消毒是必不可少的。因此，突变的Ikappabeta的过表达应废除细胞因子的作用。与野生型细胞相比，在具有腺病毒介导的突变ikappabeta转染的腺病毒介导的转染的腺病毒介导的诱导增强。还表明，通过IL-1和脂多糖转染突变的Ikappabeta，凋亡的诱导增强了。从这些结果中可以看出，nfkappabeta是凋亡的抑制剂。接下来，我们试图将腺病毒感染到肾脏转染突变的Ikappabeta，尽管注射腺病毒将腺病毒注射到仅感染的肾动脉，仅感染了近端小管。目前，我们正在开发一种将突变的Ikappabeta转移到肾脏的技术。