Mechanism of action of warming recuperating kidney medicine against the apoptosis of bone marrow stem cells

Objective:To explore the effect and mechanism of warming recuperating kidney medicine against the apoptosis of bone marrow stem cell(BMSC).Methods:The apoptosis system of SD rats BMSC cultured in vitro was established,and the apoptosis was induced by interleukin-1β(IL-1β),then the cells were respectively added with morinda root medicated serum(group A),deer-horn glue medicated serum(group B),epimedium herb medicated serum(group C),drynaria fortunei medicated serum(group D)and blank serum(group F),while group E without serum.After culturing 48 h for cells in each group,RT-PCR assay was used to detect mRNA expression levels of bcl-2 and Bax;electrophoresis method used to test the establishment of BMSC apoptosis system;and Western Blot method used to determine the protein expression levels of bcl-2 and Bax.Results:①The results of BMSC surface marker:the test results from flow cytometer were shown that CD34 and CD45 with negative expression,while CD90 with positive expression.②mRNA expression levels of bcl-2 and Bax:there was statistical difference in bcl-2mRNA expression level among all the groups(22.25±1.15,15.18±1.51,15.36±0.72,16.12±0.95,16.79±1.39,16.02±1.14;F=182.000,P=0.000).bcl-2mRNA expression level of group A was higher than that of group B,C,D,E and group F respectivrly(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000);group E was higher than group B(P=0.030).There was statistical difference in Bax mRNA expression level among all the groups(19.17±0.67,15.37±1.02,18.18±0.23,17.37±0.72,28.36±0.93,23.52±1.26;F=28.422,P=0.000).Bax mRNA expression level of group A was higher than that of group B and group D respectively(P=0.001,P=0.018),while lower than group E(P=0.001);group B was lower than group C,E and group F(P=0.013,P=0.000,P=0.000),while there was no statistical difference between group B and group D(P=0.052);group D was lower than group F(P=0.000);group E was higher than group C and group F respectively(P=0.000,P=0.000).③Electrophoresis results of bcl-2DNA and BaxDNA:there was significant difference in band between group E and group A,B,C and group D respectively through electrophoresis patterns.④Protein expression levels of bcl-2 and Bax:there was statistical difference in protein expression level of bcl-2 among all the groups(11 526.18±697.38,20 096.88±953.73,16 784.88±504.36,14 856.65±546.90,8 549.29±313.63,9 004.79±399.12;F=341.740,P=0.000).Protein expression level of bcl-2 of group A was lower than that of group B,C and group D respectively(P=0.000,P=0.000,P=0.000),while higher than that of group E and group F respectively(P=0.000,P=0.000);group B was higher than group C,D,E and group F respectively(P=0.000,P=0.000,P=0.000,P=0.000);group C was higher than group D,E and group F respectively(P=0.000,P=0.000,P=0.000);group D was higher than group E and group F respectively(P=0.000,P=0.000).There was statistical difference in protein expression level of Bax among all the groups(12 435.09±1 002.35,4 590.96±71.45,11 721.90±855.08,10 393.54±662.79,13 874.83±541.16,12 774.15±674.40;F=136.305,P=0.000).Protein expression level of Bax of group A was higher than that of group B and group D respectively(P=0.000,P=0.039);group B was lower than group C,D,E and group F respectively(P=0.000,P=0.000,P=0.000,P=0.000);group C was lower than group E(P=0.010);group D was lower than group E and group F respectively(P=0.000,P=0.002).Conclusion:The medicated serum of warming recuperating kidney medicine has the effect of inhibiting BMSC apoptosis induced by IL-1β,and its main mechanism is to inhibiting proteinase activity through up-regulation the expression of Bcl-2 while down-regulation the expression of Bax,then reduce the apoptosis rate.