| Objective:1.To use open field experiments and Morris water maze to detect the spontaneous behavior and cognitive ability of SHR rats,and observe the improvement of intestinal microorganisms,5-HT and tryptophan metabolism pathways of Jingning granules in SHR rats The expression and content of each node suggest that the upregulation of 5-HT content by Jingning granules may be a mechanism for improving the mechanism of intestinal microbial regulation of tryptophan metabolism,providing basic research and experimental basis for the development of new drugs in Jingning Formula.2.Through clinical observation of Jingning Granules in the treatment of ADHD children with qi and yin deficiency type before and after the changes in the scale scores and laboratory test results,evaluate the clinical efficacy of Jingning Granules in treating ADHD,observe Jingning Granules in the intestinal tract of children with ADHD The improvement of microbial and tryptophan metabolic pathways will provide a clinical basis for further development of major new drugs.Methods:In terms of animal experiments,50 4-week-old SHR rats were randomly divided into 5 groups,Jingning high,middle and low dose groups,tomoxetine hydrochloride group and model group,10 rats in each group.Ten Wistar rats of the same age were set as the normal group.Grouped into the stomach by drugs and saline,once a day for 6 weeks.In the fifth week,the experiment of open field and Morris water maze was carried out,and then executed.Detection of intestinal microorganisms,striatum 5-HT,5-HIAA,serum tryptophan,kynurenine content,brain stem TPH2 expression level,liver tissue TDO mRNA expression level and plasma IDO content in each group of rats.In terms of clinical research,23 children with ADHD who were admitted to the Pediatric Clinic of Dongzhimen Hospital from October 2018 to November 2019 were included,and 20 healthy controls were selected for healthy children with matching age and gender.The treatment group took Jingning granules for 8 weeks and observed the effectiveness.The TCM syndrome scores at 0,4,and 8 weeks of treatment,SNAP-IV scale scores,Conners parent questionnaire scores,and elimination test scores were conducted.Safety index detection and recording of adverse reaction events.Serum and stool were collected in the treatment group before and after treatment and in the normal control group to detect 5-HT,tryptophan,kynurenine and fecal intestinal flora.Results:Animal experiment results:1.Open field experiment results:Compared with the normal group,the exercise distance,exercise time,exercise speed,and central zone time of the model group and the static low group were significantly increased（P<0.05）,and the peripheral zone time Both were significantly shortened（P<0.05）;the exercise time and central zone time of the static high group were significantly increased（P<0.05）,The time in the peripheral area was significantly shortened（P<0.05）.Compared with the model group,the movement distance,movement speed,and time in the central area of the Western medicine group,Jinggao group and Jingzhong group were significantly reduced（P<0.05）,and the time in the peripheral area was significantly increased（P<0.05）.The exercise time of the Jingzhong group was significantly reduced（P<0.05）.2.Morris water labyrinth experiment result:Rat positioning navigation ability:Compared with the normal group,except for the model group on the first day,the remaining incubation period was significantly shortened（P<0.05）.Compared with the model group,on the 1st and 4th day,each medication group was significantly shortened（P<0.05）;on the 2nd day,except the Jinggao group,each medication group was significantly shortened（P<0.05）;on the 3rd day,the Jinggao group Both were significantly shortened（P<0.05）;on the fifth day,Jing Gao group and tomoxetine group were significantly shortened（P<0.05）.Effects of spatial learning ability and memory ability of rats in each group:compared with the normal group,the number of crossing platforms in each group was significantly increased（P<0.05）,and the residence time in the target quadrant was significantly extended（P<0.05）;except for the model group The target quadrant movement distance of each medication group was significantly increased（P<0.05）.Compared with the model group,except the static low group,the number of crossing platforms of each medication group increased significantly（P<0.05）,the target quadrant stay time was significantly extended（P<0.05）;the target quadrant movement distance of the static high group and the tomoxetine group Both were significantly increased（P<0.05）.3.Composition and difference results of intestinal flora:Venn shows that the mice in each group have the same intestinal microbes,and there are also specific flora.PCoA analysis showed that the clustering of each group was acceptable.Compared with the normal group,the model group had different bacterial composition and structure.After intervention,the Jing Gao group tended to the normal group,followed by the Jing Zhong group.Heatmap shows that the model group is different from the normal group.After intervention,the intestinal flora of Jingzhong group and Jinggao group are similar.Each group differences in species of the genus level;norankfMuribaculaceae,Alloprevotella,Lactobacillus,Ruminococcus1,Unclassified f Prevotellaceae,Prevote lla9,RuminococcaceaeUCG-014,Bacteroides,Ruminiclostridium9,RuminococcaceaeNK4 A214group,RikenellaceaeRC9gutgroup,Prevotella1,Phascolarctobacterium,PrevotellaceaeUCG-001,ChristensenellaceaeR-7group.4.Results of striatum 5-HT and its metabolites:Compared with the normal group,5-HT in the model group was significantly decreased（P<0.05）,model group,tomoxetine group,Jinggao group,Jingzhong The content of 5-HIAA in the group increased significantly（P<0.05 or P<0.01）;compared with the model group,the content of 5-HT in the tomoxetine group and Jinggao group increased significantly（P<0.05 or P<0.01）.The content of 5-HIAA in the moxetine group increased significantly（P<0.05）.5.Results of brainstem TPH2 protein expression:Compared with the normal group,the TPH2 protein expression level of the model group decreased significantly（P<0.01）;compared with the model group,each medication group increased significantly（P<0.01）.6.Results of liver tissue TDO mRNA expression:Compared with the normal group,the model group TDO mRNA expression level was significantly increased（P<0.01）;compared with the model group,each medication group was significantly decreased（P<0.01）.7.Results of serum IDO content:Compared with the normal group,the IDO content of the model group increased significantly（P<0.01）;compared with the model group,each medication group decreased significantly（P<0.01）.8.Results of serum tryptophan and kynurenine:Compared with the normal group,the content of tryptophan and kynurenine in the model group decreased significantly（P<0.05）,and the content of tryptophan in the tomoxetine group was significant Elevated（P<0.05）;the content of kynurenine in Jingzhong group and Jinglow group were significantly reduced（P<0.01）.Compared with the model group,the content of tryptophan and kynurenine in the tomoxetine group,Jinggao group,and Jingzhong group increased significantly（P<0.05 or P<0.01）.Clinical research results:1.Overall efficacy evaluation results:After treatment,the apparent efficiency was 43.48%,the effective rate was 34.78%,the ineffective rate was 21.74%,and the total effective rate was 78.26%.2.The total curative effect of TCM syndromes:After treatment,the effective rate is 78.26%,the ineffective rate is 21.74%,and the total effective rate is 78.26%.3.Results of the main syndromes of TCM syndromes:Compared with 0 weeks of treatment,the main certificates of 4 and 8 weeks of treatment were significantly improved（P<0.05 or P<0.01）;compared with 4 weeks of treatment,each of 8 weeks of treatment The main certificate of the item was significantly improved（P<0.01）.4.Results of secondary syndromes of TCM syndromes:Compared with 0 weeks of treatment,the secondary syndromes of 4 weeks of treatment,in addition to the complexion,other secondary syndromes（less sleep,more dreams,hot hands and feet,dry mouth,spontaneous sweating,night sweats,diet,stool,Tongue and veins）significantly improved（P<0.05 or P<0.01）,the 8-week treatment of secondary syndromes in addition to stool,other secondary syndromes(less sleep and dreams,hot hands and feet,dry mouth,complexion,spontaneous sweating,night sweats,diet,tongue Significantly improved（P<0.01）;compared with 4 weeks of treatment,some of the secondary syndromes（less sleep,more dreams,hand,foot and heart heat,dry mouth,spontaneous sweating,night sweats,tongue and pulse）of 8 weeks of treatment were significantly improved（P<0.01 0.01）.5.Results of SNAP-Ⅳ scale:Compared with 0 weeks of treatment,SNAP-Ⅳ scores of 4 and 8 weeks of treatment were significantly improved（P<0.01）;compared with 4 weeks of treatment,SNAP-Ⅳ scores of 8 weeks of treatment were significantly improved（P<0.01）.6.Conners scale results,compared with 0 weeks of treatment,each item of 4 and 8 weeks of treatment was significantly improved（P<0.01）;compared with 4 weeks of treatment,conduct problems,learning problems,impulsive hyperactivity and 8 weeks of treatment Hyperactivity index points improved significantly（P<0.05 or P<0.01）.7.Results of elimination test:Compared with 0 weeks of treatment,the scores of 4 and 8 weeks of treatment were significantly improved（P<0.05 or P<0.01）;compared with 4 weeks of treatment,the scores of 8 weeks of treatment were significantly improved（P<0.01）.8.Serum index:5-HT content:Compared with 0 weeks of treatment,the normal group and after treatment were significantly increased（P<0.05 or P<0.01）.Tryptophan content:Compared with before treatment,the normal group was significantly increased（P<0.05）;compared with kynurenine content,the three groups had no statistical significance.9.Intestinal flora results:Venn shows that there are many types of intestinal flora in three groups,and there are also specific flora.PCoA shows that the three groups are clusterable,the intestinal flora in the pre-treatment group is different from the normal group,and the post-treatment group tends to the normal group.There was no difference in abundance and diversity of intestinal flora among the three groups.Heatmap shows that the composition and proportion of intestinal flora before and after treatment are similar,but after treatment,it is close to the normal control group.At the door level,Firmicutes,Bacteroidetes,Proteobacteria and Verrucomicrobia decreased before treatment and Actinobacteria increased.After Jingning granule intervention,Firmicutes and Verrucomicrobia increased before treatment and Actinobacteria decreased.At the genus level,compared with the normal group,Blautia,Bacteroides,and unclassified f Lachnospiraceae decreased before treatment,and Bifidobacterium increased.After the intervention of Jingning Granules,Blautia,unclassifiedfLachnospiraceae increased,and Bifidobacterium decreased.10.During the period of taking Jingning granules,the safety indexes of children with ADHD were within the normal reference value range,and no adverse reactions occurred.Conclusion:1.Animal experiment:Jingning granule can improve the spontaneous behavior and spatial memory ability of ADHD model rats,and can improve the hyperactivity,impulsive behavior and memory learning ability of ADHD model rats;striatum of ADHD model rats 5-Insufficient HT content,Jingning granules can increase its content,preferably high and medium doses;ADHD model rats have differential flora,Jingning granules can improve their intestinal microecology;ADHD model tryptophan metabolism The pathway is abnormal.Jingning granules can play a therapeutic effect by improving the metabolism of tryptophan to promote the synthesis of striatum 5-HT;Jingning granules may increase the content of striatum 5-HT in ADHD model rats by improving the intestine The mechanism of the bacterial flora regulating tryptophan metabolism exerts its curative effect.2.Clinical research:Jingning granules can improve the clinical symptoms of children with ADHD;the structure of intestinal flora in children with ADHD is different from normal children,Jingning granules can adjust the composition and proportion of intestinal microbes in children with ADHD;serum of children with ADHD 5-The content of HT and tryptophan is lower than that of normal children,there is abnormal tryptophan metabolism of 5-hydroxytryptamine pathway,Jingning granule can improve tryptophan metabolism of 5-hydroxytryptamine pathway;Jingning granule can improve tryptophan metabolism of 5-hydroxytryptamine pathway It is through improving the mechanism of intestinal flora to regulate tryptophan metabolism to exert its curative effect. |
PDF Full Text Request