颐脑解郁复方对卒中后应激大鼠肠道菌群的影响

Effect of Yinao Jieyu Compound Formula on Intestinal Microflora in Stress Rats of Post-stroke

目的:从肠道菌群改变的角度,探索颐脑解郁复方治疗脑卒中后应激大鼠的作用机制研究。方法:将48只雄性清洁级SD大鼠随机分为假手术组、卒中后应激组、西药组和中药组各12只。假手术组只分离血管不插入线栓,卒中后应激组、西药组、中药组在大脑中动脉阻塞术后1周应用慢性束缚应激复合单笼饲养的方法制备卒中后应激大鼠模型。自脑卒中造模开始后7天,西药组给予盐酸氟西汀胶囊2.33 mg/(kg·d)灌胃,中药组给予颐脑解郁复方9.92 g/(kg·d)灌胃,其余各组大鼠灌胃10 mL/kg蒸馏水,均每日1次。分别于卒中后第14天,观察其行为学的变化,取大鼠直肠粪便,应用Illumina-MiSeq高通量测序技术检测其微生物群落结构和组成的变化。结果:与假手术组相比,卒中后应激组大鼠蔗糖水偏好度、旷场得分、游泳挣扎时间均显著降低(P <0.01,P <0.01,P <0.01),粪便中Allobaculum、Sutterella、Dorea和rc4-4比例显著下降(P <0.01,P <0.01,P <0.01,P <0.05),Oscillospira比例显著上升(P <0.01,P <0.05,P <0.01);与卒中后应激组比较,中药组大鼠旷场试验水平得分和游泳挣扎时间均显著升高(P <0.05,P <0.05),粪便中Lactobacillus和Bacteroides等6种菌属比例显著升高,其中以Lactobacillus和Bacteroides差距最为明显,分别为27.91%vs5.96%和24.21%vs9.62%(P <0.01,P <0.01),Oscillospira比例显著降低(P <0.05);与西药组相比,中药组大鼠行为学各项评分均无显著差异(均P> 0.05),粪便中Bacteroides,rc4-4比例显著升高(P <0.01,P <0.05)。结论:颐脑解郁复方可以通过增加大鼠肠道中乳杆菌属(Lactobacillus)和拟杆菌属(Bacteroides)的含量来缓解卒中后应激大鼠的类似抑郁症状。

Objective： To explore the mechanism of Yinao Jieyu Compound Formula in the treatment of post-stroke stress rats from the perspective of intestinal flora changes. Methods： A total of 48 male clean SD rats were randomly divided into 4 groups： sham-operation group, post-stroke stress group, western medicine group and traditional Chinese medicine group, with 12 rats in each group. In the sham-operation group, only the free blood vessels were not inserted into the thread embolus. The rats in post-stroke stress group, the western medicine group and the Chinese medicine group were treated as follows： after 1 week of middle cerebral artery occlusion, the rat model of PSD was prepared by using chronic restraint stress and single cage feeding. The 7th day from the beginning of the construction of the cerebral stroke model, the garage treatment began to be done. The western medicine group was given Fluoxetine Hydrochloride Capsules 2.33 mg/（kg.d）, and the Chinese medicine group was given 9.92 g/（kg· D）. The rest of the rats were gavage with 10 mL/kg distilled water, 1 time/day. The behavioral changes on the 14th day after post-stroke were observed and the microbial community structure and composition was analyzed by Illumina-MiSeq high throughput sequencing technique. Results： Compared with the sham operation group, the sucrose water preference, open field score and swimming struggle time of the post-stroke stress group were significantly decreased （P 〈 0.01, P 〈 0.01, P 〈 0.01）, and the proportions of Allobaculum, Sutterella, Dorea and rc4-4 in the feces decreased significantly （P 〈 0.01, P 〈 0.01, P 〈 0.01, P 〈 0.05）. The proportion of Oscillospira increased significantly （P 〈 0.01, P 〈 0.05, P 〈 0.01）. Compared with the post-stroke stress group, the horizontal scores and swimming struggling time of the rats in the Chinese medicine group were significantly increased （P 〈 0.05, P 〈 0.05）, and the proportions of the 6 species of Lactobacillus and Bacteroides were significantly increased in the feces. The difference between Lactobacillus and Bacteroides was the most obvious with 27.91% vs 5.96% and 24.21% vs 9.62% （P 〈 0.01, P 〈 0.01）. The proportion of Oscillospira was significantly decreased （P 〈 0.05）. Compared with the western medicine group, there was no significant difference in the behavioral scores of the rats in the traditional Chinese medicine group （P 〉 0.05）, and the proportion of Bacteroides and re4-4 in the feces was significantly increased （P 〈 0.01, P 〈 0.05）. Conclusion： Yinao Jieyu Compound Formula can alleviate similar depressive symptoms in post-stroke stress rats by increasing the content of Lactobacillus and Bacteroides in the intestine of rats.