Fibronectin-integrin signaling is required for L-glutamine's protection against gut injury.
Fibronectin-integrin signaling is required for L-glutamine's protection against gut injury.
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DOI：
10.1371/journal.pone.0050185
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wischmeyer PE
中科院分区：
文献类型：
--
作者：
Niederlechner S;Klawitter J;Baird C;Kallweit AR;Christians U;Wischmeyer PE
细胞外基质(ECM)稳定和纤维连接蛋白(FN)-整合素信号转导可介导细胞保护。L-谷氨酰胺(Gln)被认为可以防止损伤后的细胞凋亡。然而，目前尚不清楚细胞外基质稳定和FN-整合素渗透通路是否与Gln在肠道中的细胞保护机制有关。在基础和应激条件下，用谷氨酰胺加或不加FN siRNA、整合素抑制剂GRGDSP、对照多肽GRGESP或ERK1/2抑制剂PD98059和UO126处理IEC-6细胞。MTS法测定细胞存活率。Western blotting检测caspase-3、PARP、Bax、Bcl2、热休克蛋白(HSPs)、ERK1/2和转录因子HSF-1的磷酸化和/或总水平。共聚焦荧光显微镜检测细胞大小和F-肌动蛋白形态，LC-MS/MS检测细胞内谷氨酸氨基转移酶浓度。谷氨酸氨基转移酶抑制高温诱导细胞凋亡后纤维连接蛋白降解。此外，GRGDSP抑制FN-整合素的相互作用和PD98059抑制ERK1/2激酶抑制Gln的保护作用。GRGDSP可减弱Gln介导的ERK1/2磷酸化和HSF-1水平的升高。PD98059和GRGDSP在Gln治疗后也能降低HSP水平。最后，GRGDSP减弱了谷氨酰胺介导的细胞面积的增加和F-肌动蛋白组装的破坏，但对细胞内的谷氨酰胺浓度没有影响。综上所述，这些数据表明，阻止FN降解和FN整合素信号在谷氨酰胺介导的细胞保护中起着关键作用。谷氨酰胺通过FN-整合素途径传递的信号与ERK1/2和HSF-1激活诱导的HSP有关，从而减少肠损伤后的细胞凋亡。
Extracellular matrix (ECM) stabilization and fibronectin (FN)-Integrin signaling can mediate cellular protection. L-glutamine (GLN) is known to prevent apoptosis after injury. However, it is currently unknown if ECM stabilization and FN-Integrin osmosensing pathways are related to GLN’s cell protective mechanism in the intestine. IEC-6 cells were treated with GLN with or without FN siRNA, integrin inhibitor GRGDSP, control peptide GRGESP or ERK1/2 inhibitors PD98059 and UO126 under basal and stressed conditions. Cell survival measured via MTS assay. Phosphorylated and/or total levels of cleaved caspase-3, cleaved PARP, Bax, Bcl-2, heat shock proteins (HSPs), ERK1/2 and transcription factor HSF-1 assessed via Western blotting. Cell size and F-actin morphology quantified by confocal fluorescence microscopy and intracellular GLN concentration by LC-MS/MS. GLN’s prevention of FN degradation after hyperthermia attenuated apoptosis. Additionally, inhibition of FN-Integrin interaction by GRGDSP and ERK1/2 kinase inhibition by PD98059 inhibited GLN’s protective effect. GRGDSP attenuated GLN-mediated increases in ERK1/2 phosphorylation and HSF-1 levels. PD98059 and GRGDSP also decreased HSP levels after GLN treatment. Finally, GRGDSP attenuated GLN-mediated increases in cell area size and disrupted F-actin assembly, but had no effect on intracellular GLN concentrations. Taken together, this data suggests that prevention of FN degradation and the FN-Integrin signaling play a key role in GLN-mediated cellular protection. GLN’s signaling via the FN-Integrin pathway is associated with HSP induction via ERK1/2 and HSF-1 activation leading to reduced apoptosis after gut injury.
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DOI：
10.1152/ajpgi.1996.271.5.g729
发表时间：
1996-11-01
影响因子：
4.5
作者：
Goke, M;Zuk, A;Podolsky, DK
通讯作者：
Podolsky, DK
影响因子：
5.5
作者：
Hamiel, Christine R.;Pinto, Shanti;Wischmeyer, Paul E.
通讯作者：
Wischmeyer, Paul E.
影响因子：
10.6
作者：
Haussinger, Dieter;Kubitz, Ralf;Schliess, Freimut
通讯作者：
Schliess, Freimut
影响因子：
3.3
作者：
Prosser, C;Stelwagen, K;Milne, C
通讯作者：
Milne, C
影响因子：
5.5
作者：
Morrison, AL;Dinges, M;Wischmeyer, PE
通讯作者：
Wischmeyer, PE