通心络胶囊对大鼠肾缺血再灌注损伤的保护作用

Investigation of protection of Tongxinluo capsule on renal ischemical reperfusion injury

目的探讨通心络胶囊对大鼠肾缺血再灌注损伤(R IR I)的保护作用及其可能的作用机制。方法将30只雄性W istar大鼠随机分为假手术组、缺血再灌注组和缺血再灌注+通心络预处理组,各10只。缺血再灌注组和缺血再灌注+通心络预处理组采用无损伤动脉夹夹闭大鼠双侧肾蒂45 m in,再灌注24 h的方法制成急性肾缺血再灌注损伤(AR IR I)模型,其中缺血再灌注+通心络预处理组术前喂通心络7 d。假手术组仅游离双侧肾蒂,暴露60 m in后再关闭切口。光镜下观察细胞结构改变;测定血肌酐(Cr);苏木精-伊红(HE)染色观察肾小管损伤程度;免疫组化图像分析光镜下Bc l-2阳性表达;一步法TUNEL细胞凋亡检测(TUNEL法)肾小管上皮细胞中凋亡阳性细胞;激光共聚焦显示肾小管上皮细胞凋亡情况。结果缺血再灌注组和缺血再灌注+通心络预处理组Cr及肾小管损伤评分较假手术组高(P<0.05);缺血再灌注+通心络预处理组Cr及肾小管损伤评分较缺血再灌注组低(P<0.01)。假手术组肾小管上皮细胞Bc l-2有少量表达;缺血再灌注组Bc l-2在肾小管上皮细胞的表达较假手术组增加(P<0.05);缺血再灌注+通心络预处理组Bc l-2在肾小管上皮细胞的表达较缺血再灌注组增加(P<0.01)。缺血再灌注组凋亡阳性细胞数明显多于假手术组(P<0.05),多见于肾小管,尤其是远端小管细胞,少数见于近端小管,而肾间质、肾小球极少见;缺血再灌注+通心络预处理组阳性细胞数明显少于缺血再灌注组(P<0.01)。结论通心络胶囊对大鼠AR IR I具有保护作用,其作用机制可能是通过上调Bc l-2在肾小管上皮细胞的表达,抑制肾小管上皮细胞凋亡而实现的。

Objective To investigate the influence of Tongxinluo capsule（TXLC） on renal ischemic-reperfusion（I/R） injury and the possible protective mechanisms.Methods 30 male Wistar rats were randomly divided into 3 groups（n=10 in each group）： sham operation group（S group）;ischemia-reperfusion group（I/R group） and I/R＋ TXLC group（I/R＋ T group）.The models were established through clamping renal pedicles for 45 minutes following 24-hour reperfusion.TXLC（1.0 g/kg,bid） was administered intragastrically for 7 days before operation in I/R＋T group,and Sham operation group only separated double renal pedicles.Pathologic changes were examined by light microscopy.The concentration of serum creatinine was measured for the assessment of renal dysfunction.The expression of Bcl-2 was detected by immunohistochemistry image analysis;Apoptosis of renal tubular epithelial cells was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling（TUNEL） method.Results Compared with S group,the Crimea levels and score of nephric tubule injury were obviously increased in I/R group and I/R＋ T group（P〈0.05）.Compared with those in the I/R group,the Crimea levels and score of nephric tubule injury were decreased in I/R＋ T group（P〈0.01）.The expression of Bcl-2 in I/R＋ T group was obviously increased in comparison with that in I/R group（P〈0.01）.Apoptosis of renal tubular epithelial cells in I/R＋ T group was obviously increased in comparison with that in I/R group（P〈0.01）.Conclusion TXL capsule could protect the kidney from renal I/R injury by removing excess oxygen free radicals reducing the generation of MDA,and decreasing the apoptosis of renal tubuler epithelial cells.