Tianjin Key Laboratory of Acute Abdomen Disease Associated Organ Injury and ITCWM Repair, Institute of Acute Abdominal Diseases of Integrated Traditional Chinese and Western Medicine, Tianjin Nankai Hospital, Tianjin 300100, China; Nankai Clinical College, Tianjin Medical University, Tianjin 300107, China.
Department of Hepatobiliary and Pancreatic Surgery, Tianjin Nankai Hospital, Tianjin 300100, China; Nankai Clinical College, Tianjin Medical University, Tianjin 300107, China.
Biomed Pharmacother. 2020 Aug;128:110216. doi: 10.1016/j.biopha.2020.110216. Epub 2020 Jun 1.
Pancreatic stellate cells (PSCs) are the main effector cells in the development of pancreatic fibrosis. Finding substances that inhibit PSC activation is an important approach to inhibiting pancreatic fibrosis. Saikosaponin A (SSa) has numerous pharmacological activities, but its effect on PSCs remains unknown. This study was conducted to explore the effects of SSa on PSC activation in cultured rat PSCs. Cell viability, proliferation, migration and apoptosis were evaluated by MTT assays, the iCELLigence System, Transwell assays and flow cytometry. Markers of PSC activation, autophagy and the NLRP3 inflammasome were measured by real-time PCR, immunofluorescence and western blotting. Rapamycin and phenformin hydrochloride were used to determine the effect of SSa via the AMPK/mTOR pathway. The results showed that SSa suppressed PSC viability, proliferation, and migration and promoted apoptosis. SSa inhibited PSC activation, restrained PSC autophagy and suppressed the NLRP3 inflammasome. In addition, there was interaction between autophagy and the NLRP3 inflammasome during SSa inhibition of PSCs. Moreover, promotion of p-AMPK increased autophagy and the NLRP3 inflammasome. Inhibition of p-mTOR increased autophagy and decreased the NLRP3 inflammasome. Our results indicated that SSa inhibited PSC activation by inhibiting PSC autophagy and the NLRP3 inflammasome via the AMPK/mTOR pathway. These findings provide a theoretical basis for the use of SSa to treat pancreatic fibrosis and further suggest that targeting autophagy and the NLRP3 inflammasome may provide new strategies for the treatment of pancreatic fibrosis.
胰腺星状细胞（PSCs）是胰腺纤维化发展的主要效应细胞。寻找抑制 PSC 活化的物质是抑制胰腺纤维化的重要途径。柴胡皂苷 A（SSa）具有多种药理活性，但它对 PSCs 的影响尚不清楚。本研究旨在探讨 SSa 对培养的大鼠 PSCs 活化的影响。通过 MTT 检测、iCELLigence 系统、Transwell 检测和流式细胞术评估细胞活力、增殖、迁移和凋亡。通过实时 PCR、免疫荧光和 Western blot 检测 PSC 活化、自噬和 NLRP3 炎性小体的标志物。使用雷帕霉素和盐酸苯乙双胍来确定 SSa 通过 AMPK/mTOR 通路的作用。结果表明，SSa 抑制 PSC 活力、增殖和迁移，促进凋亡。SSa 抑制 PSC 活化，抑制 PSC 自噬并抑制 NLRP3 炎性小体。此外，在 SSa 抑制 PSCs 过程中，自噬和 NLRP3 炎性小体之间存在相互作用。此外，促进 p-AMPK 增加自噬和 NLRP3 炎性小体。抑制 p-mTOR 增加自噬并减少 NLRP3 炎性小体。我们的结果表明，SSa 通过 AMPK/mTOR 通路抑制 PSC 自噬和 NLRP3 炎性小体来抑制 PSC 活化。这些发现为使用 SSa 治疗胰腺纤维化提供了理论依据，并进一步表明靶向自噬和 NLRP3 炎性小体可能为胰腺纤维化的治疗提供新策略。
