目的 通过丙磺舒（PRB）抑制有机阴离子转运蛋白（OAT）1和 3，观察马兜铃酸Ⅰ（AAⅠ）对大鼠肾小管上皮细胞的急性损伤，以探究 AA Ⅰ进入肾小管上皮细胞的途径 。方法 雄 性 SD 大 鼠 随 机 分 为 空 白 对 照 组 、 溶 剂 对照 （PEG300） 组、PRB（150 mg · kg^-1）组、AA（Ⅰ 80 mg·kg^-1）组、AA（Ⅰ 80 mg·kg^-1）＋PRB（150 mg · kg^-1）组，每 2天 ig给药1次，连续给药4次。观察大鼠肾脏指数；生化仪检测血清肌酐（CREA）和尿素氮（BUN）水平；苏木精-伊红（HE）染色观察肾脏组织病理学变化；免疫组化染色观察 OAT1 和 OAT3 在肾小管的组织定位和蛋白表达水平；免疫透射电镜观察OAT1和 OAT3在肾小管上皮细胞的亚细胞定位和表达水平。结果 与溶剂对照组相比，AAⅠ组肾脏指数、CREA和 BUN水平显著增加（P＜0.01、0.001）；AAⅠ＋PRB组与AAⅠ组相比，肾脏指数、CREA和BUN水平显著降低（P＜0.05、0.001）。HE染色结果显示，与溶剂对照组相比，AAⅠ组肾近端小管上皮细胞（PCTEC）出现空泡样变性、微绒毛脱落以及片状坏死脱落，而 AA Ⅰ＋PRB 组与 AA Ⅰ组相比，PCTEC 空泡样变性率下降，无其他类型病理学变化；另外，肾远端小管上皮细胞（DCTEC）AAⅠ组与AAⅠ＋PRB组呈现少量空泡样变性。免疫组化和电镜结果显示，OAT1主要在PCTEC基底膜侧表达，OAT3主要在 DCTEC 基底膜侧表达，且 AAⅠ暴露后，与溶剂对照组比较，前者在近端小管（PCT）表达有下降趋势（P＜0.05），后者在远端小管（DCT）表达有上升趋势（P＜0.05）。结论 AAⅠ能够导致 PCT 和 DCT 的损伤，PRB 抑制 OAT1 和OAT3后，能够改善肾脏功能，并减少肾小管上皮细胞的病理学损害；OAT1可能是AAⅠ进入PCTEC的主要通道，而OAT3则可能是其进入DCTEC的主要通道。

Objective Utilizing probenecid (PRB) as an inhibitor of organic anion transporter (OAT) 1 and 3, to investigate the acute injury of renal tubular epithelial cells in rats induced by aristolochic acid I (AA Ⅰ) and elucidate the pathway through which AA Ⅰ enters renal tubular epithelial cells. Methods Male Sprague-Dawley rats were randomly assigned to five groups: the blank control group, the solvent control (PEG300) group, the PRB (150 mg·kg^-1) group, the AAⅠ (80 mg·kg^-1) group, and the AAⅠ+PRB (80 mg·kg^-1 + 150 mg · kg^-1) group. Administer ig once every two days for four consecutive doses. Observed the renal organ index of rats, and biochemical analyzer detected serum creatinine (CREA) and urea nitrogen (UREA) levels. Hematoxylin-eosin (HE) staining was employed to examine the pathological alterations in renal tissue. Immunohistochemical (IHC)staining was conducted to assess the tissue localization and protein expression levels of OAT1 and OAT3 in renal tubules. Immunoelectron microscopy was utilized to investigate the subcellular localization and expression levels of OAT1 and OAT3 in renal tubular epithelial cells. Results Compared with the solvent control group, the renal index, CREA, and BUN levels in the AA Ⅰ group were significantly increased (P <0.01, 0.001); Compared with the AAⅠ group, the renal index, CREA, and BUN levels were significantly reduced in the AA Ⅰ+PRB group (P <0.05, 0.001). HE staining revealed cellular edema, necrotic shedding, microvillous loss in proximal convoluted tubule epithelial cells (PCTEC) in the AAⅠgroup (P <0.05). Conversely, in the AAⅠ+PRB group, the incidence of vacuolar degeneration in PCTEC decreased significantly (P <0.05) without other observed pathological changes. Additionally, a minor degree of vacuolar degeneration was observed in distal convoluted tubule epithelial cells (DCTEC) in both the AA Ⅰand AA Ⅰ+PRB groups. IHC and electron microscopy revealed that OAT1 was predominantly expressed in the basolateral membrane of PCTEC, while OAT3 was primarily expressed in the basolateral membrane of DCTEC. Following exposure to AA Ⅰ, the expression of OAT1 exhibited a decreasing trend in the proximal convoluted tubule (PCT) (P <0.05), whereas the expression of OAT3 showed an increasing trend in the distal convoluted tubule (DCT) (P <0.05). Conclusion AA Ⅰ induces damage to both proximal convoluted tubules (PCT) and distal convoluted tubules (DCT), while PRB inhibiting OAT1 and OAT3 demonstrates improvement in renal function and reduction in pathological damage to renal tubular epithelial cells. These findings suggest that OAT1 may primarily facilitate the entry of AAⅠ into PCTEC, whereas OAT3 may serve as the primary route for its entry into DCTEC.

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国家自然科学基金资助项目（82160099）；贵州省科技计划项目（黔科合基础-ZK［2021］一般 356）；贵州省科技厅基础研究计划（黔科合基础-ZK［2022］一般465）