Effects of chrysophanol on hippocampal damage and mitochondrial autophagy in mice with cerebral ischemia reperfusion

Abstract

Objective

The cerebral ischemia-reperfusion (I/R) model is crucial for the study of cerebral stroke. Chrysophanol (Chry) can protect nerve damage of mice in cerebral ischemia-reperfusion injury. This study aimed at investigating the neuroprotective effects of chrysophanol through mitochondrial autophagy in mice with ischemia-reperfusion injury.

Materials and methods

Adult mice were stochastically divided into five groups: sham, I/R (solvent), I/R+Chry (dose, 10.0ml/kg), I/R+Chry (dose, 1.0ml/kg), and I/R+Chry (dose, 0.1ml/kg). The cerebral ischemia-reperfusion model was made in I/R and I/R+Chry groups. The changes in hippocampal formation were observed by hematoxylin and eosin (H&E) staining. The expressions of LC3B-II and LC3B-I protein in hippocampus were demonstrated by western blot (WB). The fluorescence intensities of NIX, LC3B, and mitochondria were detected by immunohistochemistry fluorescent (IF).

Results

Comparing with the I/R group, the I/R+Chry groups showed improvements in reducing the damage on the hippocampus, indicated by the reduced ratio of LC3B-II and LC3B-I protein, decreased fluorescence intensity of NIX and LC3B, and increased intensity of mitochondrial fluorescence.

Conclusion

Our study showed that chrysophanol may regulate mitochondrial autophagy through NIX protein and alleviate the damage of hippocampus through decreasing the level of mitochondrial autophagy.

Keywords:

• Chrysophanol
• cerebral ischemia-reperfusion injury
• hippocampus
• mitochondrial autophagy
• LC3B

Ethics statement

This study was performed following the recommendations in the Guidance for the Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of China (permit number: TCM-LAEC2014004). The experimental procedures were based on the Directive 2010/63/EU adopted by the European Union (EU), and the management and euthanasia procedures for all animals were administrated following the guidelines of Ethics Review Committee for Laboratory Animal Welfare of Hebei North University.

Author contributions

WHC, SW, and ZMQ designed the study; WHC collected the data; WHC and ZW performed the H & E Stain and IF experiments; WHC and ZMQ performed the WB experiments; SW provided experimental support; WHC and SW analyzed and interpreted the data; WHC and SW wrote the manuscript and provided financial support.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the Program for Science & Technology of Hebei Province (No. 07276166), and the Program for Innovation Talent of Hebei North University (No. 20190414), the Special Fund for graduate of Hebei North University.