血必净对脓毒性急性肾损伤大鼠肾小管细胞凋亡和相关蛋白表达的影响

目的探讨早期应用血必净对脓毒性急性肾损伤大鼠肾小管细胞凋亡，及相关蛋白B细胞淋巴瘤2（Bcl-2）和Bcl-2相关X蛋白（Bax）表达的影响。 方法54只雄性大鼠分为假手术组（Sham组）、脓毒症组（CLP组）、血必净组（CLP + XBJ组）三组，每组18只；以术后12、24和48 h作为时间观察点，将各组再分为3个亚组，每组6只。在各时间点检测大鼠的肌酐清除率（CrCl）、肾脏血流灌注量，并留取肾脏组织观察病理学变化，计算肾小管损伤评分，采用原位末端标记（TUNEL）法并通过计算积分光密度值（IOD）测定肾小管凋亡细胞，采用Western-blotting法测定肾脏髓质Bcl-2、Bax的变化。 结果三组大鼠的CrCl和肾脏血流灌注量在术后各时间点差异均有统计学意义（F = 6.405、18.821，P < 0.05）；且CLP + XBJ组在术后48 h时CrCl和肾脏血流灌注量明显高于CLP组（2.1 ± 0.5）ml/min/100 g vs.（1.1 ± 0.3）ml/min/100 g，（159 ± 38）BPU vs.（79 ± 32）BPU；P均< 0.05]。三组大鼠肾小管损伤评分比较，不同时间点差异存在统计学意义（F = 5.461，P < 0.05）；且CLP + XBJ组与CLP组比较，24、48 h时评分均有显著下降（1.6 ± 0.5）vs.（2.8 ± 0.8），（1.8 ± 0.8）vs.（3.6 ± 0.6）；P均< 0.05]。三组大鼠各时间点凋亡细胞IOD值，差异存在统计学意义（F = 7.259，P < 0.05）；CLP + XBJ组各时间点的IOD值均明显小于CLP组（26.6 ± 6.9）vs.（34.4 ± 5.0），（38.2 ± 5.3）vs.（48.0 ± 5.8），（37.6 ± 2.2）vs.（53.8 ± 6.7）；P均< 0.05]。同时，CLP + XBJ组能升高大鼠肾脏髓质Bcl-2的表达，抑制Bax的表达，维持Bcl-2/Bax的平衡，24、48 h时其Bcl-2（1.30 ± 0.09）vs.（0.76 ± 0.09），（2.12 ± 0.38）vs.（0.51 ± 0.07）；P均< 0.05]和Bax（1.19 ± 0.37）vs.（1.95 ± 0.90），（1.48 ± 0.15）vs.（2.69 ± 0.39）；P均< 0.05]的表达水平与脓毒症组相比差异均有统计学意义。 结论早期应用血必净可以减少肾小管细胞在脓毒症进程中的凋亡，改善脓毒性急性肾损伤大鼠肾脏的病理学改变，维持Bcl-2/Bax的平衡，减轻脓毒症导致的肾功能损害。

Effect of xuebijing injection on tubular apoptosis and related proteins expression in rat model of sepsis-induced acute kidney injury

ObjectiveTo investigate the effect of early application of xuebijing in rat model of sepsis-induced acute kidney injury by observing tubular apoptosis, B cell lymphoma 2 (Bcl-2) and Bcl-2 associated X protein (Bax) expressions. MethodsFifty-four male SD rats were randomly divided into three groups: Sham group (n = 18), cecal ligation and puncture (CLP) group (n = 18), and xuebijing (CLP + XBJ) group (n = 18). After rat model establishment, 12, 24, 48 h were regarded as observation time points, and each group was accordingly divided into 3 subgroups (6 rats per subgroup). Creatine clearance (CrCl), renal blood perfusion unit, renal pathological changes and renal tubular injury score were measured; renal tubular cell apoptosis was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) method and integrated optical density (IOD) was calculated. The expressions of Bcl-2 and Bax were detected by Western-blotting. ResultsThe CrCl and renal blood perfusion unit in all three groups had significant differences at each time point (F = 6.405, 18.821; P < 0.05). The CrCl and renal blood perfusion unit in the CLP + XBJ group were significantly higher than that in the CLP group (2.1 ± 0.5) ml/min/100 g vs. (1.1 ± 0.3) ml/min/100 g, (159 ± 38) BPU vs. (79 ± 32) BPU; all P < 0.05]. The renal tubular damage scores in all three groups had significant differences at each time point (F = 5.461, P < 0.05). Tubular damage scores decreased significantly in the CLP + XBJ group compared with the CLP group at 24, 48 h (1.6 ± 0.5) vs. (2.8 ± 0.8), (1.8 ± 0.8) vs. (3.6 ± 0.6); all P < 0.05]. The IOD had a significant difference in all three groups at each time point (F = 7.259, P < 0.05). In addition, the IOD in the CLP + XBJ group was much lower than that in the CLP group at each time point (26.6 ± 6.9) vs. (34.4 ± 5.0), (38.2 ± 5.3) vs. (48.0 ± 5.8), (37.6 ± 2.2) vs. (53.8 ± 6.7); all P < 0.05]. Meanwhile, xuebijing injection could increase Bcl-2 expression, inhibit Bax expression, and keep balance of Bcl-2/Bax ratio. There were significant differences of Bcl-2 (1.30 ± 0.09) vs. (0.76 ± 0.09), (2.12 ± 0.38) vs. (0.51 ± 0.07); all P < 0.05] and Bax (1.19 ± 0.37) vs. (1.95 ± 0.90), (1.48 ± 0.15) vs. (2.69 ± 0.39); all P < 0.05] in the CLP + XBJ group compared with the CLP group at 24 and 48 h. ConclusionEarly application of xuebijing might improve renal dysfunction and the pathological changes, reduce the tubular cell apoptosis, keep balance of Bcl-2/Bax ratio and relieve renal impairment in rat model of sepsis-induced acute kidney injury.