College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing, China.
Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, Chongqing, China.
PLoS One. 2024 May 29;19(5):e0303906. doi: 10.1371/journal.pone.0303906. eCollection 2024.
In this study, we aimed to investigate the protective effects of Panax notoginseng and leech (PL) on renal fibrosis and explore the mechanisms underlying their actions. For this study, we created an adenine-induced renal fibrosis model in SD rats to investigate the protective effect of PL on renal fibrosis and explore its underlying mechanism. Initially, we assessed the renal function in RF rats and found that Scr, BUN, and urine protein content decreased after PL treatment, indicating the protective effect of PL on renal function. Histological analysis using HE and Masson staining revealed that PL reduced inflammatory cell infiltration and decreased collagen fiber deposition in renal tissue. Subsequently, we analyzed the levels of α-SMA, Col-IV, and FN, which are the main components of the extracellular matrix (ECM), using IHC, RT-qPCR, and WB. The results demonstrated that PL was effective in reducing the accumulation of ECM, with PL1-2 showing the highest effectiveness. To further understand the underlying mechanisms, we conducted UPLC-MS/MS analysis on the incoming components of the PL1-2 group. The results revealed several associations between the differential components and antioxidant and mitochondrial functions. This was further confirmed by enzyme-linked immunosorbent assay and biochemical indexes, which showed that PL1-2 ameliorated oxidative stress by reducing ROS and MDA production and increasing GSH and SOD levels. Additionally, transmission electron microscopy results indicated that PL1-2 promoted partial recovery of mitochondrial morphology and cristae. Finally, using RT-qPCR and WB, an increase in the expression of mitochondrial fusion proteins Mfn1, Mfn2, and Opa1 after PL1-2 treatment was observed, coupled with a decline in the expression and phosphorylation of mitochondrial cleavage proteins Fis and Drp1. These findings collectively demonstrate that PL1-2 ameliorates renal fibrosis by reducing oxidative stress and restoring mitochondrial balance.
在这项研究中，我们旨在探讨三七和水蛭（PL）对肾纤维化的保护作用，并探讨其作用机制。为此，我们在 SD 大鼠中建立了腺嘌呤诱导的肾纤维化模型，以研究 PL 对肾纤维化的保护作用及其潜在机制。首先，我们评估了 RF 大鼠的肾功能，发现 PL 治疗后 Scr、BUN 和尿蛋白含量降低，表明 PL 对肾功能具有保护作用。HE 和 Masson 染色的组织学分析显示，PL 减少了炎性细胞浸润并减少了肾组织中胶原纤维的沉积。随后，我们使用 IHC、RT-qPCR 和 WB 分析了α-SMA、Col-IV 和 FN 的水平，这些是细胞外基质（ECM）的主要成分。结果表明，PL 有效减少 ECM 的积累，PL1-2 组效果最佳。为了进一步了解潜在机制，我们对 PL1-2 组的传入成分进行了 UPLC-MS/MS 分析。结果显示，差异成分与抗氧化和线粒体功能之间存在几种关联。这通过酶联免疫吸附测定和生化指标进一步得到证实，结果表明 PL1-2 通过减少 ROS 和 MDA 的产生以及增加 GSH 和 SOD 的水平来改善氧化应激。此外，透射电子显微镜结果表明，PL1-2 促进了线粒体形态和嵴的部分恢复。最后，使用 RT-qPCR 和 WB，观察到 PL1-2 处理后线粒体融合蛋白 Mfn1、Mfn2 和 Opa1 的表达增加，同时线粒体分裂蛋白 Fis 和 Drp1 的表达和磷酸化水平下降。这些发现共同表明，PL1-2 通过减少氧化应激和恢复线粒体平衡来改善肾纤维化。
