康复学报 (Aug 2019)
Effect of Acupuncture at ST36 on Signal Transduction Pathways of BDNF-CREB in Dentate Gyrus of Telomerase Knockout Mice
Abstract
Objective:To explore the effect of acupuncture on BDNF-CREB related signaling pathway in hippocampal dentate gyrus of Telomerase knockout mice (Terc-/-mice) and its neuro protective mechanism, in order to promote hippocampal neurogenesis, delay aging, improve learn cognitive skills, and provide possible pathways and experimental evidence.Methods:The phenotype was identified by polymerase chain reaction, and eighteen telomerase knockout mice with a length of 350 bp were selected, weighing (20±5) g. Mice were selected as the electroacupuncture group, the hand needle group and the blank group according to the random number table method, and six cases in each group. The interventional mice were fixed with a flexible plastic mesh to minimize the stress on the mice. The needles were made of homemade bipolar conjoined needles which used two 0.30 mm×13 mm needles. Wrap the needle handle with an insulating tape and insert it into the rubber transparent infusion tube. The exposed needle handle was bent, and the bent needle handle portion was connected to the Han's electro-acupuncture instrument. Han's electroacupuncture instrument was used in the electroacupuncture group to connect the bipolar conjoined needle to stimulate the bilateral ST36. The depth of the needle is about 1.5 mm. The electroacupuncture intervention parameters were continuous, 2 Hz, 2 mA, the intervention time was 30 minutes, and the intervention was performed once a day, continuous intervention for four days. During the operation, the mouse's local muscles were slightly shaken and there was no obvious struggle. The hand acupuncture group received acupuncturing at bilateral ST36, the depth of the needle was about 1.5 mm, and the twirling method was performed every five minutes (the flat-recovery method), the intervention time was 30 minutes, the intervention was performed once a day, and the intervention was continued for four days. The blank group was slightly restrained by both hands to simulate the grasping movement, no acupuncture intervention was performed during the operation, and the intervention was performed once a day at a fixed time for 30 minutes for four days. During the needle insertion and needle retention, the mice were covered with black cloth and slightly restrained by hand. After intraperitoneal injection of 10%chloral hydrate, the brain tissue was removed from the left ventricle and perfused with 4%paraformaldehyde for use. The expression levels of BDNF, pro-BDNF and CREB in hippocampal dentate gyrus of telomerase knockout mice were analyzed by immunohistochemistry.Results:The expression of pro-BDNF in the electroacupuncture group was higher than that in the hand needle group (P< 0.05). The expression of BDNF in the electroacupuncture group was higher than that in the blank group and the hand needle group (P< 0.05). The expression of CREB in the electroacupuncture group and the hand needle group was higher respectively than that in the blank group (P< 0.05).Conclusion:Acupuncture can promote the expression of BDNF, pro-BDNF and CREB in hippocampal dentate gyrus of telomerase knockout mice and exert its neurobiological effects.
