世界中医药

目的：观察益髓灸对D-半乳糖致衰老小鼠学习记忆和血清及脑、肾组织的8-羟基脱氧鸟苷酸（8-OHdG）、总抗氧化能力（T-AOC）的影响，比较2组穴位(足三里配悬钟、百会配关元)延缓脑衰老疗效，探讨其作用机制。方法：75只SPF级3月龄体质量为（30±2）g雄性昆明小鼠，水迷宫检测后随机分为5组，每组15只，即生理组、造模组、艾灸1组(足三里+悬钟穴）、艾灸2组(百会+关元穴），艾灸3组(非穴位）。造模组和各艾灸观察组予D-半乳糖按1 000 mg/（kg·d）颈背部皮下注射，同时生理组以同法注射等量的生理盐水，连续56 d。3组艾灸观察组于造模第13 d起开始艾灸，每次每穴3壮，灸单侧(或2穴)，共6壮。生理组和造模组不做治疗性干预，但给予与各观察组同时间、程度的捉抓刺激。造模、治疗结束后，再次进行水迷宫测试，将小鼠眼球取血，断头取脑,开腹取肾,制备脑、肾组织匀浆，用酶联免疫法（ELISA）检测血清及脑、肾组织8-OHdG、T-AOC水平。结果：1)水迷宫结果显示：造模、治疗前各组小鼠水迷宫各项指标及学习能力比较，差异无统计学意义(P>0.05)；造模及治疗后，5组间比较差异有统计学意义（P<0.05）。其中两两比较：a.造模及治疗后，造模组与生理组、艾灸1组、艾灸2组比较，“潜伏期、平台象比、停留时间、寻求次数、原平台象限百分比、跨越次数”6项指标比较，差异均有统计学意义（P<0.05）；b.生理组与艾灸3组比较，差异有统计学意义（P<0.05）；艾灸3组与艾灸1、2组比较，差异有统计学意义（P<0.05）。2）血清及脑、肾组织ELISA结果示：a.造模组血清和脑、肾组织8-OHdG含量显著升高，与生理组、艾灸1组和艾灸2组3组比较，差异有统计学意义(P<0.05)；生理组与艾灸3组比较，差异有统计学意义(P<0.05)，艾灸3组与艾灸1、2组比较，差异有统计学意义（P<0.05）。b.造模组血清和脑、肾组织T-AOC水平显著降低，与生理组、艾灸1组和艾灸2组3组比较，差异有统计学意义(P<0.05)；生理组与艾灸3组比较，差异有统计学意义(P<0.05)；艾灸3组与艾灸1、2组比较，差异有统计学意义（P<0.05）。结论：益髓灸防治脑衰老的作用机制可能是通过降低D-半乳糖诱导的8-OHdG表达，抑制细胞的DNA损伤，提高机体总抗氧化能力T-AOC，发挥抗氧化应激损伤的作用。

English Summary:

To observe the effects of Tonifying marrow moxibustion on learning and memory ability, expression of 8-OHdG and T-AOC in serum and cerebral and nephridial tissues in D-galactose induced aging mice, and to explore the mechanism by comparing points of the two groups (ST36 and GB39, DU20 and RN4) in the effect of delaying aging. Methods:A total of 75 SPF Kunming male mice (3 months old, weight：30±2g) were randomly and equally divided into the normal physiology group, model group, moxibustion 1 group ( ST36 and GB39), moxibustion 2 group ( DU20 and RN4) and moxibustion 3 group ( non-acupuncture) after Morris Water Maze Test. Mice in treatment and model groups were injected subcutaneously back of necks by 1000 mg/kg/day of D-galactose for 56 days, while mice in the normal physiology group were injected the same dosage of normal sodium in the same way for 56 days. Mice in the treatment groups were treated with moxibustion with 3 segments /point/time, unilateral or two points (altogether 6 segments) every day from the 13th day after molding. Mice in the normal physiology group and model group were not treated with therapeutic intervene, but to give catching stimulation for the same time and degree as the treatment groups. After modeling and treatment, all rats were tested by Morris Water Maze Test. The eyeball blood was taken, and cerebral tissue was collected by beheading. And kidney was taken by laparotomy to prepare the cerebral and kidney tissue homogenate. Expression of 8-OHdG and T-AOC in serum and the cerebral and kidney tissues were detected by ELISA method. Results:1) The Morris Water Maze results showed that, before modeling and treatment, there were no difference of learning and memory among the model and the other four groups in the indicators of Morris Water Maze Test (P>0.05); There were no significant differences among the five groups after modeling and treatment (P<0.05). Comparison between any two means showed that, 1）After modeling and treatment, there were significant difference compared with modeling group, normal physiology group, moxibustion 1 group and moxibustion 2 group, the indexes of “incubation, platform quadrant ratio, standing time, seeking time, the percentage of the original platform and crossing platform time” all had significant differences; 2）Compared with the physiology group and the moxibustion 3 group, there was significant difference (P<0.05); Compared with the moxibustion 3 group and the moxibustion 1 and moxibustion 2 group there was significant difference (P<0.05). 2) The ELISA results of the serum and cerebral-kidney tissue showed：1）the contents of 8-OHdG in the serum and cerebral-kidney tissue of model group were higher than that of the physiology group, moxibustion 1 group, moxibustion 2 group and moxibustion 3 group, which had statistic differences (P<0.05); compared with physiology group and the moxibustion 3 group, there was significant difference (P<0.05 ). 2）The contents of T-AOC in the serum and cerebral-kidney tissue of model group were significantly lower than those in the physiology group, compared with moxibustion 1 group, moxibustion 2 group and moxibustion 3 group, which had statistic differences (P<0.05); compared with the physiology group and moxibustion 3 group, there was significant difference (P<0.05); there were significant differences compared moxibustion 1 group and moxibustion 2 group with moxibustion 3 group (P<0.05). Conclusion:The mechanism of tonifying marrow moxibustion in preventing brain aging may be related to decreasing the expression of 8-OHdG which induced by D-galactose, inhibiting the damage cell DNA, increasing the content of T-AOC, and improve function of the anti-oxidative stress injury.

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