Zhongfeng Xingnao prescription alleviates injury of intracerebral hemorrhage via regulating the CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis

Abstract

Background and aim: The NLRP3 inflammasome can be activated after intracerebral hemorrhage (ICH), triggering an inflammatory response in the brain. Chinese herbal medicine Zhongfeng Xingnao Prescription (ZFXN) is commonly used in China for intracerebral hemorrhage treatment. However, the underlying treatment mechanism of it is unclear. The purpose of our study is to investigate whether ZFXN alleviates injury after intracerebral hemorrhage by blocking the activation of CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis. Experimental procedure: We evaluated the protective effect of ZFXN in a rat model of collagenase Ⅶ‐induced ICH. The neurological deficit score, cerebral hematoma-injury ratio, pathology, and ultrastructure of tissue surrounding the hematoma were evaluated after 5 days of ZFXN treatment, CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis-related protein expression around the hematoma was assessed by Western blot and immunohistochemistry. Meanwhile, ELISA measured the levels of IL-1β, IL-18, IL-6 and TNF-α in serum. Results and conclusion: After 5 days of ZFXN treatment, the score of neurological deficit and hematoma damage ratio decreased, and the cell destruction such as edema and vacuole conditions around the hematoma improved. The mechanism investigation results showed that ZFXN down-regulated expressions of CaMKII/NF-κB p65/NLRP3/GSDMD signaling axis-related protein around the hematoma area. In addition, ZFXN could attenuate the inflammatory response by regulating the activation of NLRP3 inflammasome after ICH. For the first time, we found that the efficacy of ZFXN on ICH might be related to the regulation of NLRP3 inflammasome.

Keywords

• Intracerebral hemorrhage
• NLRP3 inflammasome
• Ca2+/calmodulin–dependent protein kinase II
• Chinese herbal medicine
• Inflammation