Clinical Laboratory of Gansu Provincial Hospital of Traditional Chinese Medicine (Currently known as Gansu Provincial Hospital of TCM).
Precision Medicine Experimental Center of The First Hospital of Lanzhou University.
Chem Pharm Bull (Tokyo). 2024;72(10):923-931. doi: 10.1248/cpb.c24-00151.
Acute kidney injury (AKI) is a commonly encountered critical condition in clinical settings, often resulting from sepsis, infections or ischemia. Astragaloside IV (AS-IV) is the primary active component of Astragalus. The functions of Astragalus are mainly related to AS-IV, showing remarkable therapeutic effects in anti-inflammatory, antioxidant, immune-enhancing, and anti-tumor aspects. This study aimed to explore the role of AS-IV in AKI development. Lipopolysaccharide (LPS) was used to stimulate the HK-2 cells and rats to establish the AKI model in vivo and in vitro. After AS-IV treatment, the cell viability, pyroptosis rate, lactate dehydrogenase (LDH) activity, interleukin (IL)-18 and IL-1β contents, and cleaved-caspase-1, GSDMD-N, SIRT, FOXO3a protein levels were detected. Caspase-1 levels were analyzed by immunofluorescence staining. Additionallly, the acetylation levels of FOXO3a were detected by immunoprecipitation and Western blot assays. AS-IV treatment promoted the cell viability, and inhibited the pyroptosis, LDH activity, caspase-1 levels in the LPS stimulated HK-2 cells. AS-IV treatment decreased the IL-18 and IL-1β contents, cleaved-caspase-1 and GSDMD-N protein levels in both LPS stimulated HK-2 cells and rats. Furthermore, after EX527 treatment, a Sirtuin 1 (SIRT1) inhibitor, the role of AS-IV in the LPS stimulated HK-2 cells were reversed. AS-IV treatment increased the protein levels and decreased the acetylation levels of FOXO3a, which was reversed after EX527 treatment. Co-immunoprecipitation (CO-IP) assay and immunofluorescence staining confirmed that SIRT1 interacted with FOXO3a. In conclusion, this research demonstrated that AS-IV treatment inhibited the pyroptosis occurrence in LPS stimulated HK-2 cells and rats. This may be related to the SIRT1 mediated deacetylation of FOXO3a.
急性肾损伤（AKI）是临床常见的危重症，常由脓毒症、感染或缺血引起。黄芪甲苷（AS-IV）是黄芪的主要活性成分。黄芪的功能主要与 AS-IV 相关，在抗炎、抗氧化、免疫增强和抗肿瘤方面表现出显著的治疗效果。本研究旨在探讨 AS-IV 在 AKI 发生发展中的作用。采用脂多糖（LPS）刺激 HK-2 细胞及大鼠体内、体外建立 AKI 模型，给予 AS-IV 处理后，检测细胞活力、细胞焦亡率、乳酸脱氢酶（LDH）活性、白细胞介素（IL）-18 和 IL-1β含量及半胱氨酸天冬氨酸蛋白酶-1（caspase-1）、Gasdermin D-N（GSDMD-N）、沉默信息调节因子 2 相关酶 1（SIRT1）、叉头框转录因子 3a（FOXO3a）蛋白水平，免疫荧光染色检测 caspase-1 水平，免疫沉淀及 Western blot 检测 FOXO3a 乙酰化水平。AS-IV 处理可促进 LPS 刺激的 HK-2 细胞活力，抑制细胞焦亡、LDH 活性、caspase-1 水平。AS-IV 处理可降低 LPS 刺激的 HK-2 细胞及大鼠中 IL-18 和 IL-1β含量、caspase-1 和 GSDMD-N 蛋白水平。此外，经 Sirtuin 1（SIRT1）抑制剂 EX527 处理后，AS-IV 在 LPS 刺激的 HK-2 细胞中的作用被逆转。AS-IV 处理可增加蛋白水平并降低 FOXO3a 的乙酰化水平，经 EX527 处理后被逆转。Co-immunoprecipitation（CO-IP）assay 和免疫荧光染色证实 SIRT1 与 FOXO3a 相互作用。综上所述，本研究表明 AS-IV 处理可抑制 LPS 刺激的 HK-2 细胞及大鼠中细胞焦亡的发生，这可能与 SIRT1 介导的 FOXO3a 去乙酰化有关。
