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Attenuation of mouse mesangial cell contractility by high glucose and mannitol: Involvement of protein kinase C and focal adhesion kinase

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Journal of Biomedical Science

Abstract

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: normalD-glucose (group N), highD-glucose (group H), and control groups at the same osmolality as H plusL-glucose (group L) and mannitol (group M). Changes in the planar surface area of cells in response to 1 µM 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.

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Authors and Affiliations

  1. Graduate Institute of Medical Sciences, National Defense Medical Center, Taiwan, ROC

    Jin-Shuen Chen

  2. Graduate Institute of Life Sciences, National Defense Medical Center, Taiwan, ROC

    Shuk-Man Ka

  3. Department of Pathology, Tri-Service General Hospital, Taipei, Taiwan, ROC

    Herng-Sheng Lee, Jong-Shiaw Jin & Ann Chen

  4. Division of Nephrology, Department of Internal Medicine, Tri-Service General Hospital, No. 325, Cheng-Kung Rd., Sec. 2, Neihu 114, Taipei, Taiwan (ROC)

    Jin-Shuen Chen, Shih-Hua Lin MD & Yuh-Feng Lin MD

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Chen, JS., Lee, HS., Jin, JS. et al. Attenuation of mouse mesangial cell contractility by high glucose and mannitol: Involvement of protein kinase C and focal adhesion kinase. J Biomed Sci 11, 142–151 (2004). https://doi.org/10.1007/BF02256557

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  • DOI: https://doi.org/10.1007/BF02256557

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