目的 研究荆防颗粒对马兜铃酸I致小鼠急性肾损伤的预防和保护作用。方法 雄性C57BL/6J小鼠随机分为对照组、模型组和荆防颗粒低、中、高剂量（4、8、16 g/kg）组，各给药组连续7 d ig相应药物，第5天给药2 h后，除对照组外，其余各组ip马兜铃酸I（10 mg/kg），连续3 d，建立急性肾损伤模型。末次给药后，检测血清中肌酐（creatinine，CRE）和血尿素氮（blood urea nitrogen，BUN）水平；苏木素-伊红（HE）染色检测肾组织病理变化；采用TUNEL染色观察小鼠肾组织细胞凋亡情况；检测小鼠肾组织中丙二醛（malondialdehyde，MDA）、谷胱甘肽（glutathione，GSH）水平及超氧化物歧化酶（superoxide dismutase，SOD）活性；采用ELISA检测小鼠肾组织中单核细胞趋化蛋白-1（monocyte chemotactic protein-1，MCP-1）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）和IL-6水平；采用Western blotting检测肾组织蛋白激酶B（protein kinase B，Akt）、双微体扩增基因2（murine double minute 2，MDm2）、p53、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）蛋白表达。结果 与对照组比较，模型组小鼠血清BUN和CRE水平显著升高（P＜0.01），肾脏出现明显病理损伤；肾组织MCP-1、TNF-α、IL-1β、IL-6和MDA水平显著升高（P＜0.01），SOD活性及GSH水平显著降低（P＜0.01），提示肾脏细胞出现氧化应激损伤；肾组织中细胞凋亡数量明显增多（P＜0.01），促凋亡因子p53、Bax蛋白表达水平明显升高（P＜0.01），Akt、MDm2和Bcl-2蛋白表达受到抑制（P＜0.01）。与模型组比较，荆防颗粒组能够明显改善小鼠肾组织病理损伤，降低血清中炎症因子水平，调节氧化应激，上调肾组织Akt、MDm2和抗凋亡蛋白Bcl-2表达，抑制肾组织p53和Bax的表达，减少肾组织细胞凋亡（P＜0.01）。结论 荆防颗粒可以通过抗氧化应激和炎症来减轻马兜铃酸造成的急性肾损伤，其作用机制可能与调控Akt/MDm2/p53通路、抑制细胞凋亡有关。

Objective To study the preventive and protective effects of Jingfang Granules (荆防颗粒) on aristolochic acid I-induced acute kidney injury in mice. Methods Male C57BL/6J mice were randomly divided into control group, model group and Jingfang Granules low-, medium-and high-dose (4, 8, 16 g/kg) groups. After 2 h of administration, except the control group, the other groups were ip aristolochic acid I (10 mg/kg) for three consecutive days to establish an acute kidney injury model. After the last administration, creatinine (CRE) and blood urea nitrogen (BUN) levels in serum were detected; Hematoxylin-eosin (HE) staining was used to detect the pathological changes of renal tissue; TUNEL staining was used to observe cells apoptosis of renal tissue; Malondialdehyde (MDA), glutathione (GSH) levels and superoxide dismutase (SOD) activity in kidney tissue of mice were detected; ELISA was used to detect monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 levels in kidney tissue; Western blotting was used to detect protein kinase B (Akt), murine double minute 2 (MDm2), p53, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) protein expressions in kidney tissue. Results Compared with control group, levels of BUN and CRE in serum of mice in model group were significantly increased (P < 0.01), and kidneys had obvious pathological damage, levels of MCP-1, TNF-α, IL-1β, IL-6 and MDA in kidney tissue were significantly increased (P < 0.01), SOD activity and GSH level in kidney tissue were significantly decreased (P < 0.01), indicating that renal cells were damaged by oxidative stress; Number of apoptosis cells in renal tissue was significantly increased (P < 0.01), pro-apoptotic factors p53 and Bax protein expressions were significantly increased (P < 0.01), while Akt, MDm2 and Bcl-2 protein expressions were inhibited (P < 0.01). Compared with model group, Jingfang Granules significantly improved the pathological damage of kidney tissue of mice, reduced the level of inflammatory factors in serum, regulated oxidative stress, up-regulated the expressions of Akt, MDm2 and anti-apoptotic protein Bcl-2 in renal tissue, and inhibited p53 and Bax expressions in renal tissue, reduced the apoptosis of renal tissue cells (P < 0.01). Conclusion Jingfang Granules can alleviate acute kidney injury caused by aristolochic acid by resisting oxidative stress and inflammation, and its mechanism may be related to regulating Akt/MDm2/p53 pathway and inhibiting apoptosis.

荆防颗粒二次开发研究项目（2180071720113）