Department of General Surgery, Tianjin Medical University General Hospital, 154 Anshan Road, Heping District, Tianjin, 300052, China.
State Key Laboratory of Integrated Traditional Chinese and Western Medicine, General Hospital of Tianjin Medical University, Tianjin, 300052, China.
BMC Complement Med Ther. 2023 Feb 3;23(1):35. doi: 10.1186/s12906-023-03855-7.
Sepsis is a systemic inflammatory response syndrome caused by an infection in the body, and accompanying acute kidney injury (AKI) is a common complication of sepsis. It is associated with increased mortality and morbidity. Forsythia Fructus, the dried fruit of Forsythia suspensa (Thunb.) Vahl, is a commonly used traditional Chinese medicine.
This study aimed to elucidate the protective effect of Forsythiaside A (FTA) on sepsis-induced AKI by downregulating inflammatory and apoptotic responses, and exploring its underlying mechanism.
Septic AKI was induced through intraperitoneal injection of LPS (10 mg/kg) using male C57BL/6 mice and pretreated with FTA or control saline. First, we assessed the degree of renal injury by creatinine, blood urea nitrogen measurement, and HE staining of renal tissue; secondly, the inflammation and apoptosis were measured byELISA, qPCR, and TUNEL immunofluorescence; finally, the mechanism was explored by computer molecular docking and Western blot.
Our data showed that FTA markedly attenuated pathological kidney injuries, alleviated the elevation of serum BUN and Creatinine, suggesting the renal protective effect of FTA. Notably, FTA significantly inhibited the renal expression of proinflammatory cytokine IL-1β, IL-6, and TNF-α both at protein and mRNA levels and attenuated cell apoptosis in the kidney, as measured by caspase-3 immunoblot and TUNEL assay, indicating its anti-Inflammation and antiapoptotic properties. Mechanistically, administration of LPS resulted in robust endoplasmic reticulum (ER) stress responses in the kidney, evidenced by glucose-regulated protein 78(GRP78) upregulation, protein kinase RNA-like endoplasmic reticulum kinase (PERK) activation, eukaryotic initiation factor 2 alpha (elF2α) phosphorylation and C/EBP homologous protein (CHOP) overexpression, which could be significantly blocked by FTA pretreatment. Dynamic simulation and molecular docking were performed to provide further insight.
Collectively, our data suggest that FTA ameliorates sepsis-induced acute kidney injury via its anti-inflammation and antiapoptotic properties by regulating PERK signaling dependent ER stress responses.
败血症是一种由体内感染引起的全身炎症反应综合征，伴随的急性肾损伤（AKI）是败血症的常见并发症。它与死亡率和发病率的增加有关。连翘，连翘（Thunb.）Vahl 的干燥果实，是一种常用的传统中药。
本研究旨在通过下调炎症和凋亡反应，阐明连翘苷 A（FTA）对败血症诱导的 AKI 的保护作用，并探讨其潜在机制。
雄性 C57BL/6 小鼠腹腔注射 LPS（10mg/kg）诱导败血症性 AKI，并用 FTA 或对照生理盐水预处理。首先，我们通过肌酐、血尿素氮测量和肾组织 HE 染色评估肾损伤程度；其次，通过 ELISA、qPCR 和 TUNEL 免疫荧光测量炎症和凋亡；最后，通过计算机分子对接和 Western blot 探索机制。
我们的数据表明，FTA 显著减轻了病理性肾损伤，降低了血清 BUN 和肌酐的升高，提示 FTA 具有肾脏保护作用。值得注意的是，FTA 显著抑制了肾组织中促炎细胞因子 IL-1β、IL-6 和 TNF-α的表达，无论是在蛋白质水平还是在 mRNA 水平上，并且通过 caspase-3 免疫印迹和 TUNEL 测定抑制了肾细胞凋亡，表明其具有抗炎和抗凋亡作用。机制上，给予 LPS 导致肾脏内质网（ER）应激反应强烈，表现为葡萄糖调节蛋白 78（GRP78）上调、蛋白激酶 RNA 样内质网激酶（PERK）激活、真核起始因子 2α（elF2α）磷酸化和 C/EBP 同源蛋白（CHOP）过表达，这些反应可被 FTA 预处理显著阻断。进行了动态模拟和分子对接，以提供进一步的见解。
总之，我们的数据表明，FTA 通过调节 PERK 信号依赖的 ER 应激反应，通过其抗炎和抗凋亡特性改善败血症引起的急性肾损伤。
