Diosgenin revealed potential effect against cerebral ischemia reperfusion through HIKESHI/HSP70/NF-κB anti-inflammatory axis

Xinxin Zhang; Zhaowei Xue; Shangjun Zhu; Yuting Guo; Yan Zhang; Jianwei Dou; Jiye Zhang; Yoichiro Ito; Zengjun Guo

doi:10.1016/j.phymed.2022.153991

Diosgenin revealed potential effect against cerebral ischemia reperfusion through HIKESHI/HSP70/NF-κB anti-inflammatory axis

Phytomedicine. 2022 May:99:153991. doi: 10.1016/j.phymed.2022.153991. Epub 2022 Feb 15.

Authors

Xinxin Zhang¹, Zhaowei Xue¹, Shangjun Zhu¹, Yuting Guo¹, Yan Zhang², Jianwei Dou¹, Jiye Zhang¹, Yoichiro Ito³, Zengjun Guo⁴

Affiliations

¹ Institute of Targeted Drugs, Western China Science and Technology Innovation Harbour, Xi'an Jiaotong University, Diaotai Street, Qindou District, Xi'an, Shaanxi, China.
² Xi'an Medical University, No.1 Xinwang Road, Weiyang District, Xi'an, Shaanxi, China. Electronic address: leavies1979@aliyun.com.
³ Laboratory of Bio-separation Technologies, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
⁴ Institute of Targeted Drugs, Western China Science and Technology Innovation Harbour, Xi'an Jiaotong University, Diaotai Street, Qindou District, Xi'an, Shaanxi, China. Electronic address: guozj@mail.xjtu.edu.cn.

PMID: 35217435
DOI: 10.1016/j.phymed.2022.153991

Abstract

Background: It is a research hotspot to use natural compounds in treatment of cerebral ischemia reperfusion (I/R) for a refractory disease throughout the worldwide without available drugs or treatments at present. Our previous study has demonstrated that diosgenin (DIO), a starting material to synthesize various steroid anti-inflammatory drugs in medical industry, showed medicinal effect against I/R via inhibiting aberrant inflammatory reaction induced by I/R. However, the detailed anti-inflammatory network of DIO in treatment of I/R still remains to be further explored.

Purpose: HIKESHI was firstly identified as a novel target of DIO used for I/R by rat brain proteomic analysis, and mechanistic efforts were focused based on this gene. Hopefully, extensive detailed molecular mechanisms of DIO against I/R was established and confirmed.

Methods: The effect of DIO against I/R was examined in vitro and in vivo, which cells (SH-SY5Y and PC12) and rats were experienced to ORD/RP and MCAO exposures, respectively, to establish I/R modes. Staining was used to evaluate the pathological procedure of DIO used for I/R. Protein changes including expression, interaction, and activity during DIO's anti-I/R effect were assessed with real time PCR, western blot, Co-IP, luciferase reporter assay.

Results: In the current study, HIKESHI and HSP70 were both upregulated, when I/R cells and rats were treated with DIO in vitro and in vivo. Mechanistically, DIO stimulated the binding of HIKESHI to HSP70 and facilitated the translocation of HSP70 into nucleus. Subsequently, HSP70 blunted the transcription activity of NF-κB after physical interaction with this transcription factor, and therefore led to the suppression of its downstream pro-inflammatory cytokine (TNF-α, IL-1β, and IL-6) release into surrounding I/R lesion area. Conversely, HIKESHI or HSP70 knockdowns attenuated the nuclear translocation and restraint on NF-κB-mediated inflammation, finally resulting in the abolishment of DIO-induced anti-I/R effect. NF-κB activation also relieved the inhibitory inflammation and reversed DIO's effect against I/R, suggesting that NF-κB was the downstream target of HIKESHI and HSP70 in I/R treatment with DIO.

Conclusions: These findings established a novel HIKESHI/HSP70/NF-κB signaling pathway associated with DIO-treated I/R, which might be as therapeutic targets or drugs with potential implications for the therapeutic use of I/R in clinic.

Keywords: Cerebral ischemia reperfusion; Diosgenin; HIKESHI/HSP70/NF-κB; anti-inflammatory.