Mesangial cell hypertrophy induced by NH4Cl: role of depressed activities of cathepsins due to elevated lysosomal pH.
Enhanced ammoniagenesis is currently thought to play an important role in renal hypertrophy and subsequent tubulointerstitial fibrosis. Under certain conditions glomeruli also may be affected by ammonia toxicity. Exposure of glomeruli to augmented ammonia levels may occur: (i) in advanced liver diseases due to elevated blood ammonia concentrations; (ii) in conditions of enhanced tubular ammoniagenesis following cortical "trapping;" and (iii) due to increased ammonia formation in the glomeruli in the presence of impaired renal function. To elucidate the potential role of ammonia in glomerular injury, we investigated the effect of NH4Cl on protein turnover as well as on activities of various cathepsins in cultured rat mesangial cells. The results show that NH4Cl (20 mM) induced cell hypertrophy as defined by an increase in both cell protein content and cell volume (+38% and +10.1%, respectively, after 48 hr). This hypertrophy was associated with suppression of the activities of cathepsins B and L + B (-56.8% and -51.3% after 48 hr) and reduction of protein degradation rate (-61% after 48 hr), but without enhanced protein synthesis. Inhibition of Na+/H+ antiport by amiloride (1 mM) neither prevented the reduction of cathepsin activities nor the hypertrophy of the mesangial cells. Upon NH4Cl application lysosomal pH was elevated. This alkalinization may be causatively involved in the impairment of cathepsin B and L + B due to shifting the lysosomal pH above the optimum of their activities. In conclusion, NH4Cl induces hypertrophy but not hyperplasia in mesangial cells. This hypertrophy is caused by the reduction of protein degradation, mainly due to depressed activities of cathepsin B and L + B in the absence of enhanced protein synthesis. A shift of lysosomal pH above the optimum of the acidic cathepsins seems to be a key factor in their impaired activities in mesangial cells.
Ling
H
H
Division of Nephrology, University of Wuerzburg, Germany.
Ardjomand
P
P
Samvakas
S
S
Simm
A
A
Busch
G L
GL
Lang
F
F
Sebekova
K
K
Heidland
A
A
D000643:Ammonium Chloride / Q000494:pharmacology*; D000818:Animals; D002403:Cathepsins / Q000378:metabolism*; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects* / Q000473:pathology*; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006984:Hypertrophy; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D011506:Proteins; D000643:Ammonium Chloride; D006859:Hydrogen; D004247:DNA; D002403:Cathepsins; D010450:Endopeptidases