Effect of Tongdu Xingshen Acupuncture on Apoptosis of Nerve Cells and Expression of Nerve Growth Factor Protein in Brain Tissue of Young Rats with Cerebral Palsy

Liu ZhenhuanPan Pei Guang¹Qi Yanchao²Zhao Yong¹firewood iron³Tang Chunzhi³Wang Qinyu³Yang Junjun³Lam Kam-chuen³

1.Department of Pediatric Neurological Rehabilitation, Nanhai Obstetrics and Children's Hospital Affiliated to Guangzhou University of Chinese Medicine2.Department of Internal Medicine, Guangzhou Cancer Hospital3.College of Acupuncture and Massage, Guangzhou University of Chinese Medicine

Abstract:Objective: To observe the effect of Tongdu Xingshen acupuncture on brain nerve cell apoptosis, nerve growth factor protein expression and limb function in young rats with cerebral palsy, and to analyze whether this effect is related to acupuncture time. METHODS: The experiment was completed in the clean laboratory of Experimental Animal Center and the laboratory of Acupuncture and Massage College of Guangzhou University of Chinese Medicine from August to October 2005. ① 109 7-day-old newborn SD pups were selected. After ligating the left common carotid artery, the rats were recovered for 2 hours, placed in a transparent airtight container, incorporated into 37 ℃ constant temperature water and introduced hypoxic gas (volume fraction 0.08 oxygen and 0.92 nitrogen) at a speed of 1L/min. After 2.5 hours, the animals were taken out, and the survivors were kept warm for 1 hour for behavior measurement. Those who could not turn over, had abnormal balance or left-handed were regarded as successful models (72 rats). The remaining 24 rats were set as sham operation group: only the left common carotid artery was isolated without ligation and hypoxia treatment. Seventy-two rats with successful modeling were randomly divided into three groups: cerebral palsy rats acupuncture group I, cerebral palsy rats acupuncture group II, and model group, with 24 rats in each group. The rats were selected as Baiwill, affected side rate Gu Toujiao Sun, Neiguan, Quchi, Zusanli and Yongquan. Acupuncture group I of cerebral palsy young rats started acupuncture 24h after modeling, only acupuncture points on the limbs were added on the first 7 days, and acupuncture points on the head were added on the second day. Acupuncture was started on the 8th day after the model was made in the acupuncture group II of young rats with cerebral palsy, and the head and body needles were synchronized. Both groups were treated with acupuncture needles in Neiguan and Yongquan, without leaving needles, and then needle heads and Quchi and Zusanli. Baiwei, Ligu Toujiao Sun and Quchi and Zusanli are connected with G6805-Ⅱ electroacupuncture instrument, continuous wave, frequency 5~10Hz, duration 10min, once/d. Cerebral palsy rats acupuncture group Ⅰ continuous acupuncture 20d, cerebral palsy rats acupuncture group Ⅱ continuous acupuncture 13d. The method of acupuncture in the sham operation group was the same as that in the acupuncture group of cerebral palsy rats, and the model group was only modeled without any treatment. ③ On the 7th, 14th and 21st day after operation, the adhesive tape was stuck on the ventral side of the two forepaws of rats, and the time taken to tear off the adhesive tape was recorded. ④ On the 21st day after operation, 10 rats were randomly selected from each group for the following tests: the apoptosis of neurons in the frontal cortex and hippocampus and the expression of nerve growth factor in the hippocampus were determined by in situ end labeling and immunohistochemistry, respectively, and the number of apoptotic neurons and the expression of nerve growth factor immunoreactive cells were counted by image analyzer under light microscope (× 400). ⑤ The comparison of the rate uses the test of 2, and the comparison of the multi-means between groups uses the analysis of variance (F-q check). Results: 109 rats were included, 13 rats were lost due to model failure, and no animals died in the sham operation group. 7d after modeling, cerebral palsy rats acupuncture group Ⅰ, cerebral palsy rats acupuncture group Ⅱ, model group died 2, 5, 3; 7 ~ 14d after modeling, the three groups died 2, 2, 4; 14 ~ 21d after modeling, the three groups died 0, 0, 2. ① Neuronal apoptosis in hippocampus and frontal lobe cortex of rats: The positive cells stained by in situ end labeling in hippocampus and frontal lobe cortex of rats in the two acupuncture treatment groups were significantly reduced, and the difference was significant compared with the model group (P<0.05). And the apoptosis count of nerve cells in the ten-acupuncture I group of cerebral palsy young rats was significantly lower than that in the ten-acupuncture II group of cerebral palsy young rats (P<0.01). ② Expression of NGF protein in rat hippocampal neurons: The expression of NGF protein in the two acupuncture treatment groups was significantly enhanced, and was significantly higher or higher than that in the sham operation group and the model group in terms of quantity and intensity (P<0.01), and that in the acupuncture group I was significantly higher or higher than that in the acupuncture group II (P<0.01). ③ On the 7th day after operation, the time taken to remove adhesive tape in group I was significantly shorter than that in model group (P<0.01). On the 14th day after operation, the model group was significantly longer than the other three groups (P<0.05-0.01). On the 21st day after operation, the forelimb function of the two acupuncture groups and the model group was significantly improved, and the acupuncture group I of cerebral palsy rats was basically close to the normal level (compared with sham operation group, the difference was not obvious, P>0.05), but the model group was still significantly longer than the acupuncture group I and sham operation group (P<0.01). Conclusion: Acupuncture can inhibit the apoptosis of nerve cells in the brain of cerebral palsy rats, enhance the expression of nerve growth factor in the brain tissue of cerebral palsy rats, improve limb function, and has a certain protective effect on brain tissue injury of cerebral palsy rats, and the earlier the intervention, the better the effect.

Key words:Cerebral palsy;Brain/acupuncture therapy;Apoptosis;Nerve growth factors;Tongdu Awakening Acupuncture;

Conference Name:2011 Annual Meeting of Chinese Acupuncture and Moxibustion Society

Meeting time:2011-08-19

Venue:Beijing, China

Album:Medical and Health Science and Technology

Topic:Traditional Chinese Medicine

Classification number:R245

Tongdu Xingshen Acupuncture on Cerebral Palsy Young... Effect of Nerve Growth Factor Protein Expression_Liu Zhenhuan. pdf