推拿不同部位对孤独症谱系障碍模型鼠行为学及肠道菌群的影响OA
Effects of Tuina targeting different body parts on the behaviors and gut microflora of autistic spectrum disorder rat models
目的 探究不同部位的推拿对丙戊酸(VPA)诱导的孤独症谱系障碍(ASD)模型鼠行为学和肠道菌群的影响.方法 将 20 只怀孕 12.5 天的Sprague Dawley(SD)雌性大鼠随机分为VPA模型组[腹腔注射VPA(600 mg/kg),n=15]和生理盐水组(腹腔注射等量生理盐水,n=5).将注射生理盐水的子代雄鼠作为对照组,将注射VPA的子代雄鼠随机分为VPA组、背部组和腹部组(n=7).在出生后第 21 天,通过三箱社交、旷场实验和埋珠实验观察四组大鼠的社交能力、焦虑行为和刻板行为,并开始对背部组和腹部组进行推拿,每天 2 次,持续 14 天.第 35 天,再次进行行为学检测,并取肠道内容物进行物种组成与结构分析、标志物种与差异物种分析.结果 (1)行为学结果显示,与VPA组相比,背部组大鼠社交时间和旷场中心区运动时间显著增加(P<0.05),埋珠数量明显减少(P<0.01),说明其社交、焦虑和刻板行为得到改善;与VPA组相比,腹部组大鼠埋珠数量显著减少(P<0.05),刻板行为改善;埋珠实验中,背部组较腹部组埋珠数量更少,刻板行为改善更显著(P<0.05),而三箱社交和旷场实验两组结果无显著差异(P>0.05).(2)肠道微生物检测结果显示,在多样性分析中,与VPA组相比,背部组和腹部组肠道菌群丰富度显著增加(P<0.05),多样性增加(背部组P<0.05,腹部组P<0.01);在差异性分析中,在门水平上,与VPA组相比,腹部组厚壁菌门相对丰度呈现较为明显的降低趋势(P<0.05);在属水平上,与VPA组相比,背部组和腹部组的乳杆菌属相对丰度较VPA组显著下降(P<0.05);标志物种分析中,与VPA组相比,背部组布劳特氏菌属显著增加(P<0.05).结论 推拿能对ASD模型大鼠行为学和肠道菌群结构产生影响;推拿背部对ASD模型鼠的社交能力、焦虑行为和刻板行为疗效显著,推拿腹部仅对刻板行为有明显效果;推拿背部与推拿腹部均使ASD模型鼠肠道菌群丰富度和多样性增加,其中推拿腹部对肠道微生物多样性的改善更显著,并使物种分布更均匀.
Objective To investigate the effects of Tuina targeting different body parts on the behaviors and gut microflora of rat models with valproic acid(VPA)-induced autistic spectrum disorder(ASD). Methods Twenty female Sprague Dawley(SD)rats with 12.5 d of pregnancy were randomly divided into VPA model group[intraperitoneal injection of VPA(600 mg/kg),n=15]and saline group(intraperitoneal injection of equal amount of normal saline,n=5).The offspring male rats injected with saline were secleted as control group.The offspring male rats injected with VPA were randomly divided into VPA,dorsal,and abdominal groups(n=7 in each group).On the 21st day after birth,three-chamber social test,open field test,and marble-burying test were carried out to observe the social abilities,anxiety behaviors,and stereotypi-cal behaviors of rats in the four groups.Rats in dorsal and abdominal groups underwent Tuina for 14 d,twice a day.On the 35th day,behavioral tests were conducted again,and in-testinal contents were taken for species composition and structural analysis,as well as mark-er and differential species analysis. Results(i)According to behavioral observations,compared with VPA group,the social and movement time in the central open field of rats in dorsal group increased significantly(P<0.05),and the number of buried marbles decreased markedly(P<0.01),indicating improve-ment on their social abilities,anxiety behaviors,and stereotypical behaviors as consequences of dorsal Tuina;and the number of buried marbles was reduced as well in abdominal group when compared with VPA group(P<0.05),suggesting the improvement on their stereotypi-cal behaviors following abdominal Tuina.In the marble-burying test,the number of marbles buried in dorsal group was less than in abdominal group,and the stereotypical behaviors were improved more significantly(P<0.05),and there were no significant differences in the three-chamber social and open field tests between the two groups(P>0.05).(ii)In accor-dance with intestinal microflora detection results,compared with VPA group,both dorsal and abdominal groups showed increased richness(P<0.05)and elevated diversity(P<0.05 in dorsal group and P<0.01 in abdominal group)in intestinal microflora.The results of differen-tial analysis indicated that at the phylum level,compared with VPA group,the relative abun-dance of Firmicutes in rats in abdominal group showed a significant reduction trend(P<0.05);at the genus level,compared with VPA group,the relative abundance of Lactobacillus in rats in dorsal and abdominal groups decreased significantly(P<0.05).Dorsal group also showed significant increase in the genus Blautia in the analysis of marker species compared with VPA group(P<0.05). Conclusion Tuina impacted the behavior and gut microflora structure of ASD model rats.Dorsal intervention had a significant effect on social abilities,anxiety behaviors,and stereo-typical behaviors of ASD model rats,while abdominal intervention only had an obvious effect on stereotypical behaviors.Both dorsal and abdominal interventions increased the richness and diversity of gut microflora of ASD model rats,with abdominal intervention improving the intestinal microbial diversity more significantly and resulting in a more uniform species dis-tribution.
李桃;冯祥;智慧;黄文韬;李江山;李武
湖南中医药大学针灸推拿与康复学院,湖南长沙 410208,中国湖南中医药大学第二附属医院针灸推拿康复科,湖南长沙 410005,中国
推拿孤独症谱系障碍动物行为学肠道菌群中医外治法
TuinaAutism spectrum disorder(ASD)Animal behaviorGut microfloraTraditional Chinese medicine(TCM)External therapies
《数字中医药(英文)》 2024 (001)
90-100 / 11
Science and Technology Innovation Program of Hunan Province(2022RC1221),Hunan Provincial Health Com-mission project(D202306047422),and Chinese Medicine Research Program of Hunan Province(D2022031).
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