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
2011年09月01日
Liu Zhenhuan1Pan 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.Laboratory of Acupuncture and Massage College of 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. Select rats Baiwill, affected side rate Gu Tu Jiao Sun, Neiguan, Quchi, Zusanli, 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. The two groups were all with acupuncture needles in Neiguan and Yongquan, without leaving needles, and then needle heads and Quchi and Zusanli. Baiwill, Ligutuojiao Sun and Quchi and Zusanli are connected with G6805-Ⅱ electroacupuncture instrument, continuous wave, frequency 5~10Hz, duration 10min, once/d. Cerebral palsy rats were treated with acupuncture for 20 days in group I and 13 days in group II. The method of acupuncture in the sham operation group was the same as that in the acupuncture group I 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 x 2 test was used for comparison of rates, and the analysis of variance (F-q check) was used for comparison of multiple means between groups. Results: 109 rats were included, 13 rats were lost due to model failure, and no animals died in the sham operation group. Seven days after modeling, two, five and three rats died in the ten acupuncture group I, the acupuncture group II and the model group, respectively; seven to fourteen days after modeling, two, two and four rats died in the above three groups, respectively; and fourteen to twenty-one days after modeling, zero, two rats died in the above three groups, respectively. ① Neuronal apoptosis in hippocampus and frontal lobe cortex of rats: The number of positive cells stained by in situ end labeling in hippocampus and frontal lobe cortex of rats in the two acupuncture treatment groups was 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 hippocampal neurons of rats: 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 =, the acupuncture group of cerebral palsy rats was significantly higher or higher than that of cerebral palsy rats (P<0.01 =. ③ On the 7th day after operation, the time taken for the rats in group I to remove the adhesive tape was significantly shorter than that in the model group (P<0.01 =: on the 14th day after operation, the model group was significantly longer than that in 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 was basically close to the normal level (compared with the 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 the 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:The 7th Chinese Society of Traditional Chinese Medicine Academic Annual Meeting on External Treatment of Traditional Chinese Medicine
Meeting time:2011-09-01
Venue:Chengdu, Sichuan, China
Album:Medical and Health Science and Technology
Topic:Traditional Chinese Medicine
Classification number:R245
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