Abstract:

Objective To investigate the protective effect of active components of Jingkang Capsule on cerebral ischemia injury in rats，and to elucidate its possible mechanism. Methods Seventy SD rats were randomly divided into sham operation group， model group， and low dose （252 mg·kg?1）， medium dose （630 mg·kg?1）， and high dose of Jingkang Capsule （1 260 mg·kg?1） groups. The drugs were prepared with normal saline and administered to the rats via daily oral gavage for 14 consecutive days as preventive treatment. The rats in sham operation group and model group were given equal volumes of normal saline. After 14 days of treatment， the rats in sham operation group underwent only vessel separation without any further treatment， while the rats in other groups were subjected to cerebral ischemia model establishment using the suture occlusion method. Longa scoring method was used to assess neurological deficit in the rats in various groups. Triphenyltetrazolium chloride （TTC） staining was used to detect the cerebral infarction area in the rats in various groups. Hematoxylin-eosin （HE） staining and Nissl staining were used to observe pathomorphological changes in the brain tissues of rats in various groups. A “drug-component-target” network was constructed， and bioinformatics was used to screen the action targets of Jingkang Capsule. Western blotting method was used to detect the expression levels of protein kinase B （Akt） and apoptosis-related proteins in the cortical tissue of the rats in various groups. Results The Longa scoring method results showed that compared with sham operation group， the rats in model group showed significant neurological deficits， and the neurological deficit score was significantly increased （P<0.001）. Compared with model group， the neurological deficit scores in medium and high doses of Jingkang Capsule groups were significantly decreased （P<0.05 or P<0.01）. The TTC staining results showed that compared with sham operation group， obvious cerebral infarction areas were observed in the brain tissue of the rats in model group， and the percentage of cerebral infarction area was significantly increased （P<0.001）. Compared with model group， the percentages of cerebral infarction area of the rats in medium dose and high dose of Jingkang Capsule groups were significantly decreased （P<0.05 or P<0.01）. HE staining and Nissl staining results showed that compared with sham operation group， the brain tissue on the ischemic side of the rats in model group had looser structure with visible neuronal pyknosis， and the number of Nissl bodies in neurons was significantly decreased （P<0.001）. Compared with model group， the brain tissue on the ischemic side of rats in medium dose of Jingkang Capsule group showed reduced neuronal deformation， orderly arrangement， and relatively intact structure， and the number of Nissl bodies in neurons was significantly increased （P<0.01）. A “drug-component-target” network was constructed using drug-component-intersection target data， and the top 10 targets were screened as core targets using the cytoHubba plugin. The Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses results showed that most enriched pathways of drug-disease intersection targets were related to apoptosis and inflammation signaling pathways. The Western blotting method results showed that compared with sham operation group， the ratio of phosphorylated Akt （p-Akt）/Akt in the brain tissue of the rats in model group was significantly decreased （P<0.01）， the expression level of B-cell lymphoma 2 （Bcl-2） protein was significantly decreased （P<0.01）， and the expression levels of Bcl-2-associated X protein （Bax） and cleaved cysteinyl aspartate-specific proteinase 3 （Cleaved Caspase-3） proteins were significantly increased （P<0.01）. Compared with model group， the p-Akt/Akt ratios in the brain tissue of the rats in medium dose and high dose of Jingkang Capsule groups were significantly increased （P<0.01 or P<0.001）， the expression levels of Bcl-2 protein were significantly increased （P<0.05 or P<0.01）， and the expression levels of Bax and Cleaved Caspase-3 proteins were significantly decreased （P<0.05 or P<0.01）. Conclusion Jingkang Capsule exerts a protective effect on the brain tissue in rats with cerebral ischemia， and its protective effect may be related to the regulation of Akt and apoptosis-related protein expression.

Key words: Jingkang Capsule, Cerebral ischemia, Network pharmacology, Longa neurological behavior score, Protein kinase B