中国血液净化

中国血液净化 ›› 2025, Vol. 24 ›› Issue (06): 479-484.doi: 10.3969/j.issn.1671-4091.2025.06.008

• 基础研究 • 上一篇    下一篇

黄芪甲苷通过调控环磷酸腺苷/蛋白激酶A信号通路抑制铁死亡减轻单侧输尿管梗阻大鼠肾纤维化

贺晓丹   张紫媛   胡雅玲   方敬爱   

  1. 030001 太原，1山西医科大学第一临床医学院
    030001 太原，2山西医科大学第一医院肾内科
  • 收稿日期:2025-01-08 修回日期:2025-03-06 出版日期:2025-06-12 发布日期:2025-06-12
  • 通讯作者: 张紫媛 E-mail:ziye_402@163.com
  • 基金资助:
    国家自然科学青年基金(82405174)； 山西省基础研究计划项目(202203021222397)

Astragaloside IV alleviates renal fibrosis in unilateral ureteral obstruction rats by inhibiting ferroptosis through the regulation of the cAMP/PKA signaling pathway

HE Xiao-dan, ZHANG Zi-yuan,HU Ya-ling, FANG Jing-ai   

  1. The First Clinical Medical School of Shanxi Medical University, Taiyuan 030001, China; 2Department of Nephrology, the First Hospital of Shanxi Medical University, Taiyuan 030001, China 
  • Received:2025-01-08 Revised:2025-03-06 Online:2025-06-12 Published:2025-06-12
  • Contact: 030001 太原，2山西医科大学第一医院肾内科 E-mail:ziye_402@163.com

摘要： 目的 探究黄芪甲苷通过调控环磷酸腺苷(cyclic adenosine monophosphate，cAMP)/蛋白激酶A(protein kinase A‌，PKA)信号通路抑制铁死亡减轻单侧输尿管梗阻(unilateral ureteral obstruction，UUO)大鼠肾纤维化的相关机制。 方法 选用SPF级健康雄性SD大鼠建立UUO模型，随机分为4组(假手术组、模型组、黄芪甲苷组、氯沙坦钾组，每组6只大鼠)，手术后1天开始灌胃，连续14天。检测血清肌酐(Scr)、尿素氮(BUN)、cAMP水平，苏木精伊红(HE)染色及马松染色观察梗阻侧肾脏组织病理改变，免疫组化法检测肾组织纤维化指标[α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、纤维连接蛋白(fibronectin,FN)、I型胶原蛋白(collagen type I,COL-I)]，Western blotting法检测PKA及铁死亡相关指标[谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、血红素加氧酶-1(heme oxygenase 1,HO-1)、溶质载体家族7成员11(solute carrier family 7 member 11,xCT)]的表达。 结果 假手术组、模型组、黄芪甲苷组、氯沙坦钾组的Scr比较无统计学差异(F=2.815、P=0.065)。较假手术组，模型组的BUN升高(t=26.306，P＜0.001)；较模型组，黄芪甲苷组、氯沙坦钾组的BUN下降(t=-5.241、    -3.469，P＜0.001、P=0.002)。HE及马松染色结果显示模型组大鼠梗阻侧肾脏明显纤维化，黄芪甲苷组与氯沙坦钾组肾脏纤维化程度均较模型组轻。模型组较假手术组的α-SMA、FN、COL-I(t=17.728、9.202、13.710，均P＜0.001)、cAMP(t=9.601，P＜0.001)、PKA(t=32.321，P＜0.001)表达升高；黄芪甲苷组、氯沙坦钾组的α-SMA(t=-11.457、-5.519，P＜0.001、P=0.001)、FN(t=-6.301、-4.725，P＜0.001、  P=0.001)、COL-I(t=-6.087、-3.243，P＜0.001、P=0.012)、cAMP(t=-6.629、-5.809，均P＜0.001)、PKA(t=-22.754、-23.294，均P＜0.001)表达较模型组均降低；模型组的GPX4、HO-1、xCT较假手术组降低(t= -38.397、-41.713、-56.779，均P＜0.001)，黄芪甲苷组、氯沙坦钾组的GPX4(t=25.504、16.786，均P＜0.001)、HO-1(t=10.611、42.007，均P＜0.001)、xCT(t=7.192、3.181，P＜0.001、=0.013)表达较模型组均升高。 结论 黄芪甲苷可改善UUO大鼠肾功能，缓解肾纤维化，部分机制可能是通过调控cAMP/PKA信号通路抑制铁死亡来抑制肾纤维化。

关键词: 黄芪甲苷, 环磷酸腺苷/蛋白激酶A信号通路, 铁死亡, 肾纤维化

Abstract: Objective   To investigate the mechanism by which astragaloside IV alleviates renal fibrosis in unilateral ureteral obstruction (UUO) rats by inhibiting ferroptosis through modulation of the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling pathway.  Methods   Specific pathogen-free (SPF) male Sprague-Dawley (SD) rats were used to establish UUO models and randomly divided into four groups (n=6 per group): sham operation group, model group, AS-IV group [40 mg/(kg·d)], and losartan group [10.3 mg/(kg·d)]. Intragastric administration began one day post-surgery and continued for 14 days. Serum creatinine (Scr), blood urea nitrogen (BUN), and cAMP levels were measured. Renal histopathology was assessed via hematoxylin-eosin (HE) and Masson staining. Immunohistochemistry evaluated fibrosis markers [α-smooth muscle actin (α-SMA), fibronectin (FN), collagen type I (COL-I)], while Western blotting analyzed PKA and ferroptosis-related markers [glutathione peroxidase 4 (GPX4), heme oxygenase-1 (HO-1), solute carrier family 7 member 11 (xCT)].  Results  No significant differences in Scr were observed among groups (F=2.815, P=0.065). Compared to the sham group, the model group exhibited elevated BUN (t=26.306, P＜0.001), which was reduced in the astragaloside IV and losartan potassium groups (t=−5.241, −3.469; P＜0.001, P=0.002, respectively). HE and Masson staining revealed severe renal fibrosis in the obstructed kidneys of rats in the model group, while fibrosis was attenuated in the astragaloside IV and losartan potassium groups. Compared with the sham operation group, the model group exhibited increased expression of α-SMA, FN, COL-I (t =17.728, 9.202, 13.710, all  P＜0.001), cAMP levels (t =9.601, P＜0.001), and PKA (t =32.321, P＜0.001). In the astragaloside IV and losartan potassium groups, the expression of α-SMA (t =-11.457, -5.519, P＜0.001, P =0.001), FN (t =-6.301, -4.725, P＜0.001, P =0.001), COL-I (t =-6.087, -3.243, P＜0.001, P=0.012), cAMP levels (t =-6.629,-5.809, both P＜0.001), and PKA (t =-22.754, -23.294, both  P＜0.001) was decreased compared with the model group. Compared with the sham operation group, the model group showed decreased expression of GPX4, HO-1, and xCT (t =-38.397, -41.713, -56.779, all  P＜0.001). In the astragaloside IV and losartan potassium groups, the expression of GPX4 (t =25.504, 16.786, both P＜0.001), HO-1 (t=10.611, 42.007, both P＜0.001), and xCT (t=7.192, 3.181, P＜0.001, P =0.013) was increased compared with the model group.   Conclusions  Astragaloside IV improves renal function and mitigates fibrosis in UUO rats, potentially by suppressing ferroptosis through regulation of the cAMP/PKA signaling pathway.

Key words: Astragaloside IV, cAMP/PKA signaling pathway, Ferroptosis, Renal fibrosis

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