Effect of argatroban on brain neuronal apoptosis in rats with acute cerebral infarction

Abstract

Objective To investigate the effect of argatroban on neuronal apoptosis, and the expression of caspase-3, Bcl-2 as well as Bax in rats with acute cerebral infarction. Methods The rat model of focal cerebral ischemia was established by middle cerebral artery occlusion (MCAO) . SD rats were divided into Sham group, MCAO model group and Argatroban treatment group, and 3 time windows were set (0, 6, 12 h) . The samples of 3 time windows at 0, 6, 12 h were selected for detection, respectively. TUNEL apoptosis detection kit was used to detect neuronal apoptosis in the rat brain; Western blot and ELISA were used to detect the expression of caspase-3, Bcl-2 and Bax in the rat brain; ANOVA was used for comparison among groups, and LSD-t test was used for comparison of two groups. Results The results of TUNEL showed that the apoptosis index of the sham group and Argatroban group all decreased at 0, 6, 12 h when compared with MCAO group [0 h (14.91±4.11) ﹪ vs (2.13±0.58) ﹪, (4.22±1.01) ﹪; 6 h (24.75±3.88) ﹪ vs (3.19±0.28) ﹪, (12.14±1.90) ﹪; 12 h (48.22±2.69) ﹪ vs (5.75±1.21) ﹪, (28.45±7.84) ﹪, P< 0.05]. The results of ELISA showed that the expression of caspase-3 and Bax in rat brain neuron in MCAO group increased at 0, 6 and 12 h compared with that in the sham group, while the expression of Bcl-2 increased at 6 and 12 hours (P< 0.05) . And compared with MCAO group, the expression of caspase-3 and Bax in rat brain neuron in Argatroban group decreased at 0, 6 and 12 h, while the expression of Bcl-2 increased at 6 and 12 hours [the expression of caspase-3: 0 h (2.32±0.12) vs (1.25±0.06) 、6 h (5.91±0.60) vs (3.26±0.22) 、12 h (7.31±0.27) vs (6.42±0.18) ng/ml; the expression of Bax: 0 h (0.82±0.08) vs (0.45±0.03) 、6 h (1.00±0.05) vs (0.66±0.04) 、12 h (1.56± 0.11) vs (1.09±0. 07) ng/ml; the expression of Bcl-2: 6 h (1.07±0.08) vs (1.56±0.05) ng/ ml, 12 h (0.67±0.08) vs (1.45±0.06) ng/ml, P< 0.01]. The protein expression trends of caspase-3 and Bax in rat brain neuron among 3 groups were consistent with those of ELISA, while the expression of Bcl-2 protein in Argatroban group was higher than that in sham group and MCAO group (P< 0.01) . Conclusion Argatroban exerts neuroprotective effects by reducing neuronal apoptosis induced by acute cerebral infarction, inhibiting the expression of caspase-3 and Bax and further enhancing Bcl-2 levels. Key words: Argatroban; Cerebral infarction; Caspase-3; Bcl-2; Bax