Therapeutic effect and mechanism of mesenchymal stem cells on acute liver failure / 中华肝胆外科杂志

ABSTRACT

Objective To explore the therapeutic effects of soluble cytokines secreted by mesenchymal stem cells (MSCs) on acute liver failure (ALF).Methods MSCs isolated from Sprague-Dawley rats were determined by FACS analysis.Conditioned medium derived from MSCs (MSCs-CM) was collected and analyzed by a cytokine microarray.SD rats were divided into 3 groups:(1) ALF + dulbecco's modified eagle medium (DMEM) group:1 ml DMEM was injected into SD rats after D-Gal administration;(2) ALF + MSCs group:1 ml MSCs (1 × 106) was injected into SD rats after D-Gal administration;(3) ALF + MSCs-CM group:1 ml MSCs-CM was injected into SD rats after D-Gal administration.Biochemical indicators,survival rate,histology and inflammatory factors were studied.Exogenous recombinant rat IL-10,antirat IL-10 antibody and AG490 (STAT3 signaling pathway inhibitor) were administrated to explore the therapeutic mechanism of MSCs-CM.Results The respective serum biochemical indexes of ALF + DMEM group,ALF + MSCs group,and ALF + MSCs-CM group were:ALT (1 709.8 ± 372.1,865.5 ± 52.8,964.7 ± 414.6 U/L),AST (4234.0 ± 807.3,2440.8 ± 511.9,2739.8 ± 587.3 U/L),andTBil (79.3 ± 10.9,43.8 ± 7.0,61.2 ± 6.7 μg/L).The survival rates of the three groups were 10.0％,80.0％,and 70.0％,respectively.The levels of inflammatory factors in each group were IFN-γ (69.8 ± 4.7,46.4 ± 4.3,54.6 ± 2.4pg/ml),IL-1β (58.5 ± 7.6,40.5 ± 6.9,44.1 ± 6.0pg/ml),IL-6 (71.9 ± 16.1,38.4 ± 7.7,45.3 ± 9.0),and IL-10 (38.3 ± 6.0,75.4 ± 11.1,59.6 ± 11.9 pg/ml).Protein microarray results suggested that MSCs-CM expresses a variety of inflammatory-related cytokines,with IL-10 levels being most pronounced.IL-10 (ALT 1 126.9 ± 419.3 U/L,AST2370.8 ± 561.2 U/L) alone significantly reduced transaminase levels compared with ALF group (ALT 1 709.8 ± 372.1 U/L,AST 4234.0 ± 807.3 U/L),while anti-IL-10 antibody (ALT 1 568.5 ± 325.4 U/L,AST4043.7 ± 819.0 U/L) neutralized the therapeutic effect of MSCs-CM (ALT 964.7 ± 414.6 U/L,AST 2 739.8 ± 587.3 U/L).IL-10 could significantly increase the level of pSTAT3 in ALF rats (0.93 ± 0.03 vs 0.68 ± 0.01),while STAT3 inhibitor AG490 (0.84 ± 0.04) could decrease the expression of pSTAT3 and reverse the therapeutic effect of IL-10.Conclusion The factors released by MSCs,especially IL-10,have the potential therapeutic effect on ALF,and STAT3 signaling pathway may mediate the anti-inflammation effects of IL-10.