Expression levels and significance of cysteinyl aspartate-specific protease-3 and Beclin1 in abdomi-nal trauma rats
Objective To investigate the expression levels and significance of cysteinyl aspartate-specific protease(Caspase)-3 and Beclin1 in abdominal trauma rats.Methods A total of 20 SD rats were divided into control group and abdominal trauma group by a digital table method.Caspase-3 and Beclin1 in the intestinal tissues of the two groups were analyzed by fluorescence quantitative polymerase chain reaction(PCR)and Western blotting,and the apoptosis level of the intestinal tissues of the two groups was analyzed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The number of LC3 points in intestinal tissues of the two groups was analyzed by immunofluorescence.The expression of cleaved Caspase-3 protein in the intestinal tissues of the two groups was analyzed by immunohistochemistry.The levels of intestinal inflammatory factors were detected by enzyme linked immunosorbent assay(ELISA).T test was used to compare measurement data between groups.Results The mRNA expression levels of Caspase-3 and Beclin1 in the intestinal tissue of rats in the control group(0.96±0.11,1.08± 0.12)were significantly lower than those in the abdominal trauma group(1.82±0.12,1.77±0.18;t=16.910,10.120,P＜0.05).The expression level of inflammatory factor tumor necrosis factor-α(TNF-α)in intestinal tissue of rats in control group[(18.82±3.56)pg/ml]was significantly lower than that of rats in abdominal trauma group[(71.98±11.80)pg/ml,t=13.640,P＜0.05].The relative expression lev-els of Caspase-3 and Beclin1 in intestinal tissue of rats in the control group(0.60±0.08,0.89±0.08)were significantly lower than those in the abdominal trauma group(1.34±0.18,1.56±0.16;t=12.170,12.230,P＜0.05).The relative expression level of intestinal cleaved Caspase-3 protein in the control group(97.66±11.25)was significantly lower than that in the abdominal trauma group(191.33±24.47,t=10.990,P＜0.05).The positive rate of TUNEL staining in intestinal tissue of rats in control group[(2.83±1.10)％]was significantly lower than that in abdominal trauma group[(50.59±10.15)％,t=14.790,P＜0.05].The number of LC3 points in intestinal tissue of rats in control group(45.20±14.91)was significantly less than that in abdominal trauma group(82.81±13.17,t=5.977,P＜0.05).West-ern blotting results showed that the expression level of autophagy substrate p62 in the intestinal tissue of rats in the control group(1.26±0.11)was significantly higher than that of rats in the abdominal trauma group(0.62±0.11,t=12.940,P＜0.05).Conclusion The expression levels of Caspase-3 and Beclin1 in intestinal tissues were significantly increased after abdominal trauma,which led to a significant increase in the level of apoptosis and autophagy of intestinal cells and aggravated intestinal injury.
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