基于脑-肠轴探讨调胃承气汤治疗脑缺血模型大鼠作用机制研究

Exploration of the Mechanism of Tiaowei Chengqi Decoction(调胃承气汤) on Cerebral Ischemia Model Rats Based on Brain-gut Axis

目的探讨调胃承气汤对脑缺血模型大鼠的治疗作用及机制。方法脑缺血模型采用改良线栓法进行制备,于造模24 h后根据Longa评定表对大鼠的神经功能缺损程度进行评估,将造模成功的24只大鼠随机分为模型组、阳性药组(双歧杆菌四联0.4 g/mL)、调胃承气汤组(4.7 g/mL),另设假手术组。治疗组予对应药物灌胃,模型组及假手术组予等容积的无菌蒸馏水灌胃,连续10 d。造模后的1 d、10 d进行神经功能缺损评分;于末次给药后采用16S rRNA基因测序法检测大鼠盲肠内容物的肠道菌群多样性改变情况,酶联免疫法测定脂多糖(LPS)、D-乳酸(D-Lac)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)的表达。结果(1)与假手术组相比,造模后各组大鼠mNSS评分显著升高(P<0.01);与模型组相比,两治疗组均能下调mNSS评分,且调胃承气汤组大鼠的mNSS评分下降最为显著(P<0.01)。(2)与假手术组相比,模型组大鼠菌群OTU丰富度、Alpha多样性均降低,而调胃承气汤组大鼠肠菌的丰富度及多样性均明显增加。与假手术组相比,模型组大鼠门水平下厚壁菌门Firmicutes和变形菌门Proteobacteria增多,拟杆菌门Bacteroide减少;给药10 d后调胃承气汤组大鼠的上述菌门均有回调。(3)与假手术组相比,模型组LPS、D-Lac、TNF-α、IL-1β含量显著增加(P<0.01),调胃承气汤能显著下调上述指标水平(P<0.01)。结论调胃承气汤对脑缺血模型大鼠脑损伤的保护作用可能与增加肠道菌群多样性及调节肠道菌群产物以减轻脑组织炎症反应有关。

Objective To investigate the therapeutic effect and mechanism of Tiaowei Chengqi Decoction(调胃承气汤)on cerebral ischemia model rats.Methods The cerebral ischemia model was prepared by modified wire embolization method,and the neurological impairment of the rats was evaluated according to the Longa evaluation table 24 h after modeling.The 24 rats were randomly divided into model group,positive drug group(bifidobacteria quadrupleted 0.4 g/m L),Tiaowei Chengqi Decoction group(4.7 g/m L),and sham operation group.The treatment group was given corresponding drugs by intragastric administration,and the model group and the sham operation group were given equal volume of sterile distilled water by intragastric administration for 10 consecutive days.Neurological deficits were scored 1 d and 10 d after modeling.After the last administration,16S r RNA gene sequencing was used to detect the changes of intestinal microflora diversity in cecal contents of rats,and the expressions of LPS,D-LAC,TNF-α and IL-1β were determined by enzyme-linked immunosorbent assay.Results(1)Compared with sham operation group,m NSS score of rats in each group was significantly increased after modeling(P<0.01);Compared with model group,m NSS score in both treatment groups could be down-regulated,and m NSS score in Tiaowei Chengqi Decoction group decreased most significantly(P<0.01).(2)Compared with sham operation group,OTU richness and Alpha diversity of intestinal bacteria in model group were decreased,while the richness and diversity of intestinal bacteria in Tiaowei Chengqi Decoction group were significantly increased.Compared with sham operation group,Firmicutes and Proteobacteria increased and Bacteroidetes decreased in model group at phylum level.10 days after administration of Tiaowei Chengqi Decoction,the above mentioned microphyla were all recalled.(3)Compared with sham operation group,the contents of LPS,D-LAC,TNF-α and IL-1β in model group were significantly increased(P<0.01),and Tiaowei Chengqi Decoction could significantly decrease the levels of these indexes(P<0.01).Conclusion The protective effect of Tiaowei Chengqi Decoction on brain injury of cerebral ischemia model rats may be related to increasing the diversity of intestinal flora and regulating the products of intestinal flora to alleviate brain inflammation.