Effect of electroacupuncture at zusanli and futu acupoints on expression of apoptosis factor caspase-3 in spinal cord injury rats / 中国组织工程研究

ABSTRACT

BACKGROUND: Traditional Chinese medicine has proposed the theory of “treatment for flaccidity aims at Yangming meridian” in the Internal Canon of Medicine. However, there have been relatively few reports on electroacupuncture at the stomach channel of Foot Yangming in the treatment of spinal cord injury. OBJECTIVE: To investigate the effect of electroacupuncture stimulation of Zusanli and Futu on Caspase-3 expression in injured segments of spinal cord injury rats. METHODS: Sixty-four Sprague-Dawley female rats, SPF grade, were randomly divided into four groups, i.e., control group, pre-labeling group, post-injury labeling group and electroacupuncture group, with 16 rats in each group. The control group and the pre-labeling group were intraperitoneally given Brdu (50 mg/kg) for 10 days before the operation. The control group was given a bite to remove the lamina on the 11th day, which did not damage the spinal cord. A model of spinal cord injury was made in the pre-labeling group on the 11th day. The post-injury labeling group and the electroacupuncture group were injected intraperitoneally with Brdu for 10 days after the spinal cord injury was made, and the post-injury labeling group was not treated. The electroacupuncture group started electroacupuncture at Zusanli and Futu acupoints on the 3rd day after the model was established. Spinal cord specimens were taken at 3, 10, 17, and 24 days after injury in each group, and the changes in the number of neurons were observed by hematoxylin-eosin staining. The expression of Brdu positive cells was observed by immunohistochemistry. The changes in Caspase-3 mRNA and protein expression were detected by qRT-PCR and western blot, respectively. The study protocol was approved by the Animal Ethic Committee of Guangxi Medical University, with an approval No. 201712001. RESULTS AND CONCLUSION: The expression of Caspase-3 mRNA and protein after spinal cord injury was higher than that of the control group without spinal cord injury, and the number of neurons after spinal cord injury was significantly lower than that of the control group. With the passage of time, the mRNA and protein expression of Caspase-3 gene in the pre-labeling, post-injury labeling and electroacupuncture groups increased first and then decreased, while the Caspase-3 expression in the electroacupuncture group decreased significantly, which was significantly different from that in the pre-labeling and post-injury labeling groups (P < 0.05). Hematoxylin-eosin staining results showed that the number of neurons in the electroacupuncture group was significantly higher than that in pre-labeling and post-injury labeling groups, and gradually approached the number of neurons in the control group on the 24th day. Immunohistochemical results showed that the positive protein expression in the electroacupuncture group increased first and then decreased, reaching the highest on the 17th day and the lowest on the 24th day. The overall positive protein expression was significantly higher than that in the other groups (P < 0.05). To conclude, electroacupuncture stimulation of Zusanli and Futu acupoints can reduce the expression of Caspase-3 mRNA and protein, reduce the apoptosis of nerve cells, and promote the number of neurons, thus promoting the regeneration and repair of nerve cells.