Yunnan Characteristic Plant Extraction Laboratory, Key Laboratory of Medicinal Chemistry for Natural Resource, Ministry of Education and Yunnan Province, School of Chemical Science and Technology, Yunnan University, Kunming 650500, PR China; Chinese Materia Medica, Yunnan University of Chinese Medicine, Kunming 650500, PR China.
State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences Kunming 650201, PR China.
Phytomedicine. 2023 Sep;118:154958. doi: 10.1016/j.phymed.2023.154958. Epub 2023 Jul 8.
As one of the most commonly used folk medicines in "Dai" ethno-medicine system, Alstonia scholaris (l.) R. Br. has also been used for treat "water related diseases", such as chronic kidney disease. However, few study was reported for it on the intervention of chronic glomerulonephritis (CGN).
To investigate the effect and potential mechanism of indole alkaloids from A. scholaris leaves in ICR mice with adriamycin nephropathy, as well as providing experimental evidence for the further application.
ICR Mice were selected for injections of adriamycin (ADR) to induce the CGN model and administered total alkaloids (TA) and four main alkaloids continuously for 42 and 28 days, respectively. The pharmacological effects were indicated by serum, urine, and renal pathological observations. The targets and pathways of indole alkaloids on CGN intervention were predicted using the network pharmacology approach, and the immortalized mice glomerular podocyte (MPC5) cells model stimulated by ADR was subsequently selected to further verify this by western blotting and RT-qPCR methods.
TA and four major compounds dramatically reduced the levels of urinary protein, serum urea nitrogen (BUN), and creatinine (CRE) in ADR - induced CGN mice, while increasing serum albumin (ALB) and total protein (TP) levels as well as ameliorating kidney damage. Moreover, four alkaloids effected on 33 major target proteins and 153 pathways in the CGN, among which, PI3K-Akt as the main pathway, an important pathway for kidney protection by network pharmacology prediction, and then the four target proteins - HRAS, CDK2, HSP90AA1, and KDR were screened. As a result, Val-and Epi can exert a protective effect on ADR-stimulated MPC5 cells injury at a concentration of 50 μM. Furthermore, the proteins and RNA expression of HRAS, HSP90AA1, and KDR were down-regulated, and CDK2 was up-regulated after the intervention of Val-and Epi, which were supported by Western blotting and RT-qPCR. Additionally, Val-and Epi inhibited ROS production in the MPC5 cells model.
This study is the first to confirm the potential therapeutic effect of alkaloids from A. scholaris on CGN. TA with major bioactive components (vallesamine and 19‑epi-scholaricine) could exert protective effects against the ADR-induced CGN by regulating four key proteins: HRAS, CDK2, HSP90AA1, and KDR of the PI3K-Akt pathway.
作为傣医药体系中最常用的民间药物之一，垂茉莉（Alstonia scholaris（L.）R. Br.）也被用于治疗“水相关疾病”，如慢性肾病。然而，关于其对慢性肾小球肾炎（CGN）的干预作用，鲜有研究报道。
探讨垂茉莉叶中的吲哚生物碱对阿霉素肾病 ICR 小鼠的作用及可能机制，为进一步应用提供实验依据。
选择 ICR 小鼠注射阿霉素（ADR）诱导 CGN 模型，分别连续给予总碱（TA）和 4 种主要生物碱 42 天和 28 天。通过血清、尿液和肾脏病理观察来评价药效。采用网络药理学方法预测吲哚生物碱对 CGN 干预的靶点和通路，随后选择阿霉素刺激的永生化小鼠肾小球足细胞（MPC5）细胞模型，通过 Western blot 和 RT-qPCR 方法进一步验证。
TA 和 4 种主要化合物显著降低了 ADR 诱导的 CGN 小鼠尿蛋白、血清尿素氮（BUN）和肌酐（CRE）水平，同时提高了血清白蛋白（ALB）和总蛋白（TP）水平，改善了肾脏损伤。此外，4 种生物碱作用于 CGN 的 33 个主要靶蛋白和 153 个通路，其中 PI3K-Akt 作为主要通路，是网络药理学预测的肾脏保护的重要通路，随后筛选出 4 个靶蛋白——HRAS、CDK2、HSP90AA1 和 KDR。结果表明，在 50 μM 浓度下，Val 和 Epi 对 ADR 刺激的 MPC5 细胞损伤具有保护作用。此外，Val 和 Epi 干预后 HRAS、HSP90AA1 和 KDR 的蛋白和 RNA 表达下调，CDK2 表达上调，Western blot 和 RT-qPCR 结果支持这一结论。此外，Val 和 Epi 抑制了 MPC5 细胞模型中的 ROS 产生。
本研究首次证实了垂茉莉生物碱治疗 CGN 的潜在疗效。TA 及其主要生物活性成分（瓦来萨明和 19-表-垂茉莉碱）可通过调节 PI3K-Akt 通路中的 4 个关键蛋白（HRAS、CDK2、HSP90AA1 和 KDR）对 ADR 诱导的 CGN 发挥保护作用。
