摘要
BACKGROUND： Postoperative cognitive dysfunction （POCD） is very common in elderly patients. Some Chinese agents for eliminating oxygen-derived free radicals and reducing oxidative stress injury can obviously decrease the error times of the animal models of Alzheimer disease in the water maze test. However, whether they can be used to treat POCD or not is unclear. OBJECTIVE： To observe the changes of learning and memory abilities and the levels of intedeukin-6 （IL-6） and tumor necrosis factor alpha （TNF-α） in aging rats after ketamine-induced anesthesia, and investigate the effects of Fufang Shexiang injection which can eliminate oxygen-derived free radicals. DESIGN ： A randomized controlled animal study SETTING ： Experimental animal center of Guangzhou University of Traditional Chinese Medicine MATERIALS ： Twenty-eight SD aging rats （20 months old, 350-400 g, male or female） randomly divided into 4 groups with 7 rats in each group： control group, ketamine group, ketamine＋low-dosage Fufang Shexiang group and ketamine＋high-dosage Fufang Shexianggroup. Ketamine （Shanghai Zhongxi Pharmaceutical Co., Ltd., China, 50 mg/mL, 2 mL/ampule, Batch No. 040207）; Fufang Shexiang injection （Jiling Weiwei Pharmaceutical Co., Ltd., China, 2 mL/ampule, Batch No. 20031203） was composed of musk, curcumae, camphol, acon graminei rhizome, agastache rugosa, menthol, bezoar, comu bubali, scutellaria, gardenia, oncha margaritifera usta, etc. Enzyme-linked immunoabsorbent assay （ELISA） kit （Jingmei Bio-Tech Co., Ltd, Beijing, Batch No. 20050915）. Morris water maze （Upwards Teksystems Co.,Ltd., Beijing）. METHODS ： The experiment was carded out in the experimental animal center of Guangzhou university of Traditional Chinese Medicine From September to October in 2005. （1） Grouping and interventions： In the control group, the rats were intraperitoneally injected with saline （4 mL/kg per day） at 1-6 days; In the ketamine group, ketamine＋low and high-dosage Fufang Shexiang groups, the rats were intraperitoneally injected with ketamine on the 1= day, saline （4 mL/kg per day） and Fufang Shexiang （2 and 6 mL/kg per day） were intraperitoneally injected at 2-6 days respectively. （2）There were two parts for the Morris water maze test： Place navigation test： It lasted for 6 days. The rats swam freely for 2 minutes before anesthesia on the 1= day （DO）; Rats were placed into from a fixed position to locate the platform after 1 hour of drug injection. The latency to find the platform was recorded manually with the second stop-clock. The shorter the latency, the better the leaming ability. Spatial probe test： At D5, after the last navigating experiment, the platform was removed away. The rats were placed into the water facing against the wall to find the platform again, times of swimming through the quadrant which the platform had ever been in 2 minutes would be recorded. The more the times or the longer the duration, the better the memory. （3） Determinations of IL-6 and TNF-α in peripheral blood： At D5, IL-6 and TNF-α levels in peripheral blood were measured by ELISA. （4） Statistical analysis： The t test （within groups） or one-way analysis of variance （between groups） were applied.MAIN OUTCOME MEASURES ： （1） The escape latency; （2） The times of swimming through the hidden platform quadrant after it was removed; （3） The levels IL-6 and TNF-α in peripheral blood. RESULTS ： All the 28 rats were involved in the final analysis. （1） Escape latency： As time prolonged, the latencies in the control group and ketamine ＋high-dosage Fufang Shexiang group were gradually shortened （P 〈 0.05）. The latencies in the ketamine group were all longer than those in the control group except that on the 3^nd day （P〈 0.05）. The latency in the ketamine group on the 4^th day was obviously longer than that in the ketamine ＋high-dosage Fufang Shexiang group [（1.7±0.3）, （0.9±0.7） minutes, P 〈 0.05]. The latencies in the ketamine group on the 2^nd〈 to 5^th days＋low-dosage Fufang Shexiang group were close to those in the control group （P 〉 0.05）, whereas that on the 6^th day was obviously longer than that in the control group [（0.9±0.7）, （0.3±0.3） minutes, P〈 0.05]. The latencies in the ketamine ＋high-dosage Fufang Shexianggroup were all close to those in the control group at each time point （P〉 0.05）. （2） Swimming times through the quadrant of the hidden platform within 2 minutes after it was removed： The swimming times in the ketamine group and ketamine ＋low-dosage Fufang Shexianggroup were obviously fewer than those in the control group [（2.7±2.1）, （2.7±3.2）, （4.7±1.8）, （5.3±3.8） times, P〈 0.05]; Whereas there were no significant differences between the ketamine group and ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）, as well as between the ketamine group and ketamine ＋high-dosag Fufang Shexiang group （P 〉 0.05）. （3） Changes of IL-6 and TNF-α levels in peripheral blood： The IL-6 level in the control group was obviously lower than those in the ketamine group and ketamine ＋low-dosage Fufang Shexiang group [（101.08±58.73）, （161.12±115.77）, （346.42±184.76） ng/L, P〈 0.05], but had no obvious difference as compared with that in the ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）; The IL-6 level in the ketamine ＋high-dosage Fufang Shexiang group was not significantly different from those in the ketamine group and ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）. The TNF-（x in the control group was obviously lower than those in the other three groups [（53.64±31.4）, （96.69±79.34）, （151.66±37.50）, （141.50±106.85） ng/L, P 〈 0.05], and there were no significant differences among the latter three groups （P 〉 0.05）.CONCLUSION： Abilities of learning and memory in aging rats were impaired induced by anesthesia with intraperitoneal injection of ketamine, also accompanied by the increases of IL-6 and TNF-α. Fufang Shexiang injection can ameliorate the impairments, especially that of higher dosage.
BACKGROUND： Postoperative cognitive dysfunction （POCD） is very common in elderly patients. Some Chinese agents for eliminating oxygen-derived free radicals and reducing oxidative stress injury can obviously decrease the error times of the animal models of Alzheimer disease in the water maze test. However, whether they can be used to treat POCD or not is unclear. OBJECTIVE： To observe the changes of learning and memory abilities and the levels of intedeukin-6 （IL-6） and tumor necrosis factor alpha （TNF-α） in aging rats after ketamine-induced anesthesia, and investigate the effects of Fufang Shexiang injection which can eliminate oxygen-derived free radicals. DESIGN ： A randomized controlled animal study SETTING ： Experimental animal center of Guangzhou University of Traditional Chinese Medicine MATERIALS ： Twenty-eight SD aging rats （20 months old, 350-400 g, male or female） randomly divided into 4 groups with 7 rats in each group： control group, ketamine group, ketamine＋low-dosage Fufang Shexiang group and ketamine＋high-dosage Fufang Shexianggroup. Ketamine （Shanghai Zhongxi Pharmaceutical Co., Ltd., China, 50 mg/mL, 2 mL/ampule, Batch No. 040207）; Fufang Shexiang injection （Jiling Weiwei Pharmaceutical Co., Ltd., China, 2 mL/ampule, Batch No. 20031203） was composed of musk, curcumae, camphol, acon graminei rhizome, agastache rugosa, menthol, bezoar, comu bubali, scutellaria, gardenia, oncha margaritifera usta, etc. Enzyme-linked immunoabsorbent assay （ELISA） kit （Jingmei Bio-Tech Co., Ltd, Beijing, Batch No. 20050915）. Morris water maze （Upwards Teksystems Co.,Ltd., Beijing）. METHODS ： The experiment was carded out in the experimental animal center of Guangzhou university of Traditional Chinese Medicine From September to October in 2005. （1） Grouping and interventions： In the control group, the rats were intraperitoneally injected with saline （4 mL/kg per day） at 1-6 days; In the ketamine group, ketamine＋low and high-dosage Fufang Shexiang groups, the rats were intraperitoneally injected with ketamine on the 1= day, saline （4 mL/kg per day） and Fufang Shexiang （2 and 6 mL/kg per day） were intraperitoneally injected at 2-6 days respectively. （2）There were two parts for the Morris water maze test： Place navigation test： It lasted for 6 days. The rats swam freely for 2 minutes before anesthesia on the 1= day （DO）; Rats were placed into from a fixed position to locate the platform after 1 hour of drug injection. The latency to find the platform was recorded manually with the second stop-clock. The shorter the latency, the better the leaming ability. Spatial probe test： At D5, after the last navigating experiment, the platform was removed away. The rats were placed into the water facing against the wall to find the platform again, times of swimming through the quadrant which the platform had ever been in 2 minutes would be recorded. The more the times or the longer the duration, the better the memory. （3） Determinations of IL-6 and TNF-α in peripheral blood： At D5, IL-6 and TNF-α levels in peripheral blood were measured by ELISA. （4） Statistical analysis： The t test （within groups） or one-way analysis of variance （between groups） were applied.MAIN OUTCOME MEASURES ： （1） The escape latency; （2） The times of swimming through the hidden platform quadrant after it was removed; （3） The levels IL-6 and TNF-α in peripheral blood. RESULTS ： All the 28 rats were involved in the final analysis. （1） Escape latency： As time prolonged, the latencies in the control group and ketamine ＋high-dosage Fufang Shexiang group were gradually shortened （P 〈 0.05）. The latencies in the ketamine group were all longer than those in the control group except that on the 3^nd day （P〈 0.05）. The latency in the ketamine group on the 4^th day was obviously longer than that in the ketamine ＋high-dosage Fufang Shexiang group [（1.7±0.3）, （0.9±0.7） minutes, P 〈 0.05]. The latencies in the ketamine group on the 2^nd〈 to 5^th days＋low-dosage Fufang Shexiang group were close to those in the control group （P 〉 0.05）, whereas that on the 6^th day was obviously longer than that in the control group [（0.9±0.7）, （0.3±0.3） minutes, P〈 0.05]. The latencies in the ketamine ＋high-dosage Fufang Shexianggroup were all close to those in the control group at each time point （P〉 0.05）. （2） Swimming times through the quadrant of the hidden platform within 2 minutes after it was removed： The swimming times in the ketamine group and ketamine ＋low-dosage Fufang Shexianggroup were obviously fewer than those in the control group [（2.7±2.1）, （2.7±3.2）, （4.7±1.8）, （5.3±3.8） times, P〈 0.05]; Whereas there were no significant differences between the ketamine group and ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）, as well as between the ketamine group and ketamine ＋high-dosag Fufang Shexiang group （P 〉 0.05）. （3） Changes of IL-6 and TNF-α levels in peripheral blood： The IL-6 level in the control group was obviously lower than those in the ketamine group and ketamine ＋low-dosage Fufang Shexiang group [（101.08±58.73）, （161.12±115.77）, （346.42±184.76） ng/L, P〈 0.05], but had no obvious difference as compared with that in the ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）; The IL-6 level in the ketamine ＋high-dosage Fufang Shexiang group was not significantly different from those in the ketamine group and ketamine ＋low-dosage Fufang Shexiang group （P 〉 0.05）. The TNF-（x in the control group was obviously lower than those in the other three groups [（53.64±31.4）, （96.69±79.34）, （151.66±37.50）, （141.50±106.85） ng/L, P 〈 0.05], and there were no significant differences among the latter three groups （P 〉 0.05）.CONCLUSION： Abilities of learning and memory in aging rats were impaired induced by anesthesia with intraperitoneal injection of ketamine, also accompanied by the increases of IL-6 and TNF-α. Fufang Shexiang injection can ameliorate the impairments, especially that of higher dosage.
