槐耳清膏通过抑制ROS介导的腺泡细胞焦亡减轻小鼠急性胰腺炎的严重程度

Huaier alleviates acute pancreatitis in mice by reducing ROS-induced pyroptosis in acinar cells

目的探究槐耳清膏对急性胰腺炎(acute pancreatitis,AP)的治疗作用及其潜在机制。方法体内利用雨蛙肽诱导的小鼠AP模型验证槐耳清膏(简称槐耳)的治疗效果,采用HE染色及免疫组化染色评价小鼠胰腺组织的病理改变、采用透射电镜观察小鼠胰腺的焦亡形态;体外以266-6细胞系作为实验载体验证槐耳对腺泡细胞的保护作用,利用电镜及Western blotting评价腺泡细胞的焦亡水平,利用ROS荧光探针检测腺泡细胞的氧化应激状态。结果槐耳治疗明显改善了小鼠AP的严重程度,HE染色表现为胰腺内腺泡坏死和炎细胞浸润减少等,同时伴有血清淀粉酶水平下降;而免疫组化染色和Western blotting发现,槐耳治疗有效抑制小鼠胰腺组织内焦亡相关分子NLRP3、GSDMD等的表达;进一步的电镜观察发现槐耳治疗降低炎症状态下小鼠胰腺腺泡细胞的焦亡水平;此外,体外实验发现,槐耳干预能显著降低胰腺腺泡细胞266-6的ROS水平,且ROS介导的腺泡细胞焦亡能被槐耳有效抑制。结论槐耳能有效减轻AP的严重程度,这一效应是通过抑制ROS介导的胰腺腺泡细胞焦亡所实现的。

Objective To investigate the therapeutic effect of Huaier on acute pancreatitis(AP) and its potential mechanism.Methods A mouse model of cerulean-induced AP was used to verify the therapeutic effect of Huaier in vivo.HE staining and immunohistochemical staining were used to evaluate the histopathological changes of the pancreas,and transmission electron microscopy was used to observe the pyroptosis morphology of the pancreas.In vitro,266-6 cell line was used as the experimental carrier to verify the protective effect of Huaier on acinar cells.Electron microscopy and Western blotting were used to evaluate the pyroptosis level of acinar cells,and ROS fluorescence probe was used to detect the oxidative stress state of acinar cells.Results Huaier significantly alleviated the severity of AP in mice.HE staining of pancreas showed that necrosis and inflammatory cell infiltration were reduced,and the level of serum amylase was decreased.Immunohistochemical staining and Western blotting showed that Huaier effectively inhibited the expressions of pyroptosis-related molecules such as NLRP3 and GSDMD in pancreatic tissue.Electron microscopy showed that Huaier could reduce the pyroptosis level of pancreatic acinar cells under inflammatory state.In addition,the level of ROS in acinar cells was significantly reduced after the intervention of Huaier,and ROS-mediated pyroptosis of acinar cells could be effectively inhibited by Huaier.Conclusion Huaier can effectively reduce the severity of AP by inhibiting ROS-mediated pyroptosis of acinar cells.