Protective effect of ceftriaxone on hippocampal neurons in subarachnoid hemorrhage rats and its mechanism / 吉林大学学报(医学版)

ABSTRACT

Objective: To investigate the therapeutic effect of ceftriaxone in the rats with subarachnoid hemorrhage (SAH), and to clarify its mechanism. Methods: A total of 48 adult male SD rats were randomly divided into sham operation group, SAH group, 3-methyl adenine (3-MA) group and ceftriaxone (CEF) group; there were 12 rats in each group. The SAH models were established by intravascular puncture; the rats were administered intraperitoneally; the dose of 3-MA was 15 mg · kg-1, and the dose of CEF was 500 mg · kg-1. The rats were sacrificed 24 h after model establishment, and the pathology of brain tissue of the rats was observed by HE staining. The apoptosis of neurons was detected by TUNEL staining. The number of autophagy-related proteins Beclin-1 and LC3-II and the apoptosis factor Caspase-3 protein in hippocampus tissue of the rats in various groups were detected by immunohistochemistry and Western blotting methods. Results: Compared with SAH group, the neurological score of the rats in CEF group was significantly increased (P<0. 01). The HE staining results showed the nerve cells in the hippocampus area of the rats in SAH group were swollen with loose cytoplasm and had obvious nuclear condensation, fragmentation and dissolution; compared with SAH group, the neural degeneration of the rats in CEF group was improved, and more normal nerve tissue morphology appeared; while the nerve cell damage in 3-MA group was more serious, the dead nerve cells were in full filed, and the cell layers were disordered. The quantitative analysis results showed that the number of necrotic nerve cells in CEF group was significantly lower than that in SAH, and the number of necrotic nerve cells in 3-MA group was significantly higher than that in SAH group (P< 0. 05). The immunohistochemical staining results showed that the number of Beclin-1 and LC3-II posivive cells in hippocampus tissue of the rats in CEF group was significantly higher than that in SAH group (P<0. 05); the number of Beclin-1 and LC3-II positive cells in 3-MA group was significantly lower than that in SAH group (P< 0. 05). The TUNEL staining resluts showed that the number of apoptotic nerve cells in CEF group was significantly lower than that in SAH group (P<0. 05), and the number of apoptotic nerve cells in 3-MA group was higher than that in SAH group (P<0. 05). The Western blotting results showed that the expression level of Caspase-3 protein in hippocampus tissue of the rats in CEF group was significantly lower than that in SAH group (P<0. 05), while the expression level of Caspase-3 protein in 3-MA group was significantly higher than that in SAH group (P< 0. 05). Conclusion: CEF has the protective effect on the nerve injury of the rats with SAH, and its mechanism may be related to up-regulation of autophagy and reduction of apoptosis of the neurons.