当归补血颗粒对大鼠肾缺血再灌注损伤的保护机制研究

Protective Mechanism of Danggui Buxue Granules on Renal Ischemia Reperfusion Injury in Rats

关键词:  当归补血颗粒  肾微透析技术  免疫组化法  肾缺血再灌注损伤

Key words:  Danggui Buxue granules  renal microdialysis technique  immunohistochemistry  renal ischemia reperfusion injury

摘要:

目的 研究当归补血颗粒对大鼠肾缺血再灌注损伤的保护作用，探究可能的保护机制。方法 采用无创动脉夹阻断肾蒂血管构建大鼠肾缺血再灌注损伤模型，肾微透析技术结合HPLC测定腺苷及其代谢产物含量；免疫组化法测定肾组织Bax、Bcl-2、iNOS蛋白表达。结果 假手术组腺苷、次黄嘌呤和肌苷的含量分别为(0.57±0.11)，(3.14±0.20)，(0.16±0.03)μmol·L^-1，保持稳定并作为各实验组基准值，肾缺血再灌注损伤后90 min，模型组升高至(8.61±0.62)，(10.92±1.14)，(0.85±0.05)μmol·L^-1，而当归补血颗粒组升高至(6.91±0.67)，(6.04±0.67)，(0.61±0.13)μmol·L^-1，明显低于模型组(P<0.05)。免疫组化显示，与假手术组比较，肾缺血再灌注损伤后各组大鼠肾组织细胞中Bcl-2、Bax、iNOS蛋白表达显著增强(P<0.01)。再灌注后，当归补血颗粒组与模型组同时间点比较Bcl-2蛋白表达明显增强(P<0.05)，而Bax和iNOS蛋白表达明显减弱(P<0.05)。结论 当归补血颗粒对大鼠肾缺血再灌注损伤的保护机制可能与抑制ATP降解，上调Bcl-2基因表达，下调Bax、iNOS基因表达有关。

Abstract:

OBJECTIVE To investigate the protective effect of Danggui Buxue granules and search for the possible protective mechanism in rats. METHODS Using noninvasive arterial clamp to block the renal pedicle vascular to induce renal ischemia-reperfusion injury in rats and applying the kidney microdialysis technology to collects kidney dialysis fluid, then the content of adenosine and its metabolites was measured by HPLC. Making use of immunohistochemistry method to explore protein expression of the Bax, Bcl-2, iNOS. RESULTS The concentration of adenosine, hypoxanthine and inosine of model group raised from (0.57±0.11), (3.14±0.20), (0.16±0.03)μmol·L^-1 to (8.61±0.62), (10.92±1.14), (0.85±0.05)μmol·L^-1, respectively after renal ischemia reperfusion injury, but Danggui Buxue granules group(DG group) raised up to (6.91±0.67), (6.04±0.67), (0.61±0.13)μmol·L^-1, which were lower than the model group significantly. The results of immunohistochemistry showed that compared with sham operation group(SO group), after renal ischemia-reperfusion injury each group kidney tissues and cells of the Bax, Bcl-2 and iNOS protein expression significantly enhanced(P<0.01). However, after reperfusion, the Bcl-2 protein expression of DG group was higher than IR group significantly (P<0.05), while the Bax and iNOS protein expression of DG group was lower than IR group significantly (P<0.05). CONCLUSION The results showed that protective mechanism of Danggui Buxue granules on renal ischemia reperfusion injury in rats may be related to inhibiting ATP degradation, and attributed to up-regulation of Bcl-2 gene expression and down-regulation of Bax and iNOS gene expression.