College of First Clinical Medical, Shandong University of Traditional Chinese Medicine, Jinan, China.
Department of Nephrology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
J Ethnopharmacol. 2023 Feb 10;302(Pt A):115882. doi: 10.1016/j.jep.2022.115882. Epub 2022 Oct 29.
Heidihuang Wan (HDHW) is a classic Chinese herbal formula, which was first recorded in the "Suwen Bingji Qiyi Baoming Collection" written by Liu Wansu during the Jin Dynasty (1115-1234 AD). It is commonly used clinically for the treatment of kidney diseases and its curative effect is stable. Previous animal experiments have confirmed that HDHW can effectively improve renal fibrosis. However, the underlying pharmacological mechanism remains unclear.
Renal tubular epithelial cell (RTEC) apoptosis is one of the main pathological features of renal fibrosis. This study aimed to observe the effect and underlying mechanism of HDHW on the apoptosis of RTECs to further explore the pathological mechanism of HDHW against renal fibrosis.
We examined the HDHW composition in rat serum. In vitro, we first screened out the optimal intervention concentration of HDHW on RTECs using the MTT assay. Hypoxia/reoxygenation was then used to induce apoptosis of RTECs (H/R-RTECs), which were divided into H/R-RTEC, astragaloside IV (positive control), HDHW, and RTECs groups. After 48 h of drug intervention, apoptosis of RTECs was detected using flow cytometry and protein expression was detected by western blotting. The 5/6 nephrectomy rat model was constructed and divided into the normal control, 5/6 nephrectomy, HDHW, and astragaloside IV groups. After 8 weeks of treatment, TUNEL staining was used to detect cell apoptosis, and western blotting was used to detect protein expression.
HDHW downregulated the expression of pro-apoptotic proteins Bax and Caspase3, up-regulated the expression of anti-apoptotic protein Bcl-2, activated the PI3K/Akt/mTOR signaling pathway, and reversed the early apoptosis of RTECs, thereby resisting the apoptosis of RTECs.
HDHW inhibits apoptosis of RTECs by modulating the PI3K/Akt/mTOR signaling pathway. This study provides experimental evidence for the anti-fibrotic effect of HDHW on the kidneys and partially elucidates its pharmacological mechanism of action.
还魂地黄丸（HDHW）是一种经典的中草药配方，最早记录于金代（公元 1115-1234 年）刘完素所著的《素问病机气宜保命集》中。它在临床上常用于治疗肾脏疾病，疗效稳定。先前的动物实验已经证实，HDHW 可以有效改善肾纤维化。然而，其潜在的药理机制尚不清楚。
肾小管上皮细胞（RTEC）凋亡是肾纤维化的主要病理特征之一。本研究旨在观察 HDHW 对 RTEC 凋亡的影响及其潜在机制，以进一步探讨 HDHW 治疗肾纤维化的病理机制。
我们检测了大鼠血清中的 HDHW 成分。在体外，我们首先通过 MTT 检测筛选出 HDHW 对 RTEC 最佳的干预浓度。然后使用缺氧/复氧（H/R）诱导 RTEC 凋亡（H/R-RTECs），将其分为 H/R-RTEC、黄芪甲苷（阳性对照）、HDHW 和 RTECs 组。药物干预 48 h 后，通过流式细胞术检测 RTEC 凋亡，通过 Western blot 检测蛋白表达。构建 5/6 肾切除大鼠模型，并分为正常对照组、5/6 肾切除组、HDHW 组和黄芪甲苷组。治疗 8 周后，通过 TUNEL 染色检测细胞凋亡，Western blot 检测蛋白表达。
HDHW 下调了促凋亡蛋白 Bax 和 Caspase3 的表达，上调了抗凋亡蛋白 Bcl-2 的表达，激活了 PI3K/Akt/mTOR 信号通路，逆转了 RTEC 的早期凋亡，从而抑制了 RTEC 的凋亡。
HDHW 通过调节 PI3K/Akt/mTOR 信号通路抑制 RTEC 凋亡。本研究为 HDHW 抗肾纤维化提供了实验依据，并部分阐明了其药理作用机制。
