Attenuation of Mouse Mesangial Cell Contractility by High Glucose and Mannitol: Involvement of Protein Kinase C and Focal Adhesion Kinase

Jin Shuen Chen, Herng Sheng Lee, Jong Shiaw Jin, Ann Chen, Shih Hua Lin, Shuk Man Ka, Yuh Feng Lin

研究成果: 雜誌貢獻文章同行評審

20 引文 斯高帕斯(Scopus)

摘要

Hyperglycemia and mannitol activate protein kinase C (PKC) and induce mesangial cell hypocontractility that subsequently may modulate renal function. Since focal adhesion kinase (FAK) activation is known to be linked with PKC activity, FAK may also be involved in mesangial cell contraction. To facilitate our understanding of the PKC- and FAK-modulating mechanism, we developed an in vitro model of mouse mesangial cell hypocontractility induced by hyperglycemia or mannitol. Mouse mesangial cells (CRL-1927) were exposed to: normal D-glucose (group N), high D-glucose (group H), and control groups at the same osmolality as H plus L-glucose (group L) and mannitol (group M). Changes in the planar surface area of cells in response to 1 IμM phorbol 12-myristate 13-acetate (PMA) were determined. Western blot analyses for PKC, phosphorylated (p)-PKC, tyrosine phosphorylation, FAK, and p-FAK were done on each of these four groups. The effects of mannitol in various doses on cell contraction and activation of PKC and FAK were also assayed. The planar surface areas of groups H and M both showed an attenuated change in response to PMA stimulation. Before PMA stimulation, the baseline PKC expression of groups H and M showed a higher expression of p-PKCα and δ than that seen in group N (p <0.05). Results of tyrosine phosphorylation and immunoprecipitation showed that FAK may be involved in this contraction process. The total amount of FAK showed no significant difference among the four experimental groups; however, p-FAK was found to have significantly increased in group M (p <0.05). The use of PKC and tyrosine kinase inhibitors reduced PMA-induced mesangial cell contraction in all four groups. Activation of PKCα, δ, and FAK with the resultant inhibition of mesangial cell contraction by mannitol was found to be dose-dependent. These results may provide a correlation between increased expression of several PKC isoforms and, in particular, increased phosphorylation levels of PKCα and δ and hypocontractility induced by high glucose and mannitol treatment. Furthermore, the mannitol-induced hypocontractility involving PKC and FAK occurred in a dose-dependent manner.
原文英語
頁(從 - 到)142-151
頁數10
期刊Journal of Biomedical Science
11
發行號2
DOIs
出版狀態已發佈 - 2004
對外發佈

ASJC Scopus subject areas

  • 生物化學、遺傳與分子生物學 (全部)

指紋

深入研究「Attenuation of Mouse Mesangial Cell Contractility by High Glucose and Mannitol: Involvement of Protein Kinase C and Focal Adhesion Kinase」主題。共同形成了獨特的指紋。

引用此