| Background and objective: Decreased renal function in aging is mainly due to the dysfunction of renal component cells, so investigation of the alternation of glumerular mesangial cell phynotypes and functions have great values in understanding the mechanism of renal senescence. Connexin 43 and gap junction intercellular communication (GJIC) have an important role in a variety of cellular processes including cell differentiation and growth control, but until now, there have been no reports regarding the alternation of cx43 expression and GJIC in aging mesangial cells and their contribution to mesangial cell senescence. In this study, we examined the phenotype characters and connexin 43 expression and GJIC of mesangial cells in aging rats. Then, we established mesangial cell senescence model accelerated by high glucose, used RNA interference and gene transfer techniques to knockdown or overexpress connexin 43 gene in rat mesangial cells, so as to determined the contributes of connexin 43 to mesangial cell senescence.Method: The mesangial cells from 28- month-old and 3-month-old rats were cultured, cell proliferation ability , P27, P21 expression and SA-P-gal staining were measured. We used real time PCR, Northern blot, Western blot, immunocytochemistry method to detect the gene and protein expression of connexin43, fluorescence recovery after photobleaching assay (FRAP) to investigate GJIC function of mesangial cells frome different age rats. Then, we investigated the aging-related phenotype alternation of mesangial cells cultured in high glucose and the influence of high glucose on connexin 43 expression and GJIC. Furthermore, we used RNA interference techniques to knockdown connexin 43 gene expression, used gene transfer techniques to get connexin 43 gene overexpression , to determined whether the changes of connexin 43 activity were associated with mesangial cells senescence under high glucose.Result: The proliferation ability were significantly decreased in mesangial... |
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