ABSTRACT
OBJECTIVE To investigate the regulatory effects of couplet medicinals of Atractylodes macrocephala-Aucklandia lappa on gut microbiota and short-chain fatty acids （SCFAs） in the diarrhea-type irritable bowel syndrome （IBS-D） rats with spleen deficiency. METHODS The IBS-D rat model with spleen deficiency was induced by intragastric administration of Senna alexandrina combined with restraint stimulation. The model rats were divided into model group， positive control group （pinaverium bromide 1.5 mg/kg）， A. macrocephala-A. lappa low-dose， medium-dose and high-dose groups （0.7， 1.4， 2.8 g/kg）， with 6 rats in each group. Another 6 healthy rats were taken as the blank control group. The blank control group and the model group were given normal saline intragastrically， and other groups were given relevant drug liquid intragastrically， once a day， for consecutive 14 days. The general characteristics of rats and fecal water content were observed， and intestinal sensitivity [evaluating by abdominal wall withdrawal reflex （AWR） threshold] and the intestinal propulsion rate were determined. The serum levels of 5- hydroxytryptamine（5-HT）and SP were detected， and the pathological changes of colon tissue were observed； the protein expressions of 5-HT-3 receptor（5-HT3R）， 5-HT4R and 5-HT transporter（SERT） in colon tissue of rats were detected. 16S rRNA sequencing was performed for the feces of rats in blank control group， model group and A. macrocephala-A. lappa high-dose group； the contents of acetic acid， propionic acid and butyric acid in the feces of the rats were determined. RESULTS Compared with the model group， the body weight after 7 and 14 days of medication， fecal water content， AWR threshold， and the protein expressions of 5-HT4R and SERT in colon tissue were increased significantly in the A. macrocephala-A. lappa medium-dose and high-dose groups （P＜0.05 or P＜0.01）； serum contents of 5-HT and SP， intestinal propulsion rate （except for A. macrocephala-A. lappa medium-dose group）， the protein expression of 5-HT3R in colon tissue were decreased significantly （P＜0.01）； diarrhea relief， mental state recovery， and partially recovery of the structure of colon tissue were all found； moreover， the diversity and species number of gut microbiota were reduced in A. macrocephala-A. lappa high-dose group and the content of butyric acid in fecal samples was significantly reduced （P＜0.05）. CONCLUSIONS The compatibility of A. macrocephala and A. lappa can improve intestinal motility and sensitivity of IBS-D model rats with spleen deficiency， and alleviate diarrhea. This may be related to improving changes in intestinal microbiota structure， reducing 5-HT expression and butyric acid content， and increasing 5-HT4R and SERT expression.
ABSTRACT
ObjectiveTo investigate the effect and mechanism of Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex in regulating the intestinal function in the rat model of slow transit constipation （STC） due to yang deficiency via the vasoactive intestinal peptide （VIP）/cathelicidin antimicrobial peptide （cAMP）/protein kinase A （PKA）/aquaporin （AQP） pathway. MethodSD rats were randomized into 6 groups （n=6）， including a control group， a model group， high-， medium-， and low-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex groups， and a prucalopride group. Other groups except the control group were treated with loperamide hydrochloride combined with ice water by gavage for the modeling of STC due to yang deficiency. The number of fecal pellets， time to the first black stool defecation， fecal water content， intestinal propulsion rate， and score of fecal properties were recorded in each group. At the end of the treatment， the colon was stained with hematoxylin-eosin （HE） to reveal the histopathological changes and Alcian blue/periodic acid-Schiff （AB-PAS） to reveal the secretion of colonic mucus. The enzyme-linked immunosorbent assay （ELISA） was employed to measure the level of VIP in the serum. The mRNA level of AQP in the colon was measured by polymerase chain reaction （Real-time PCR）. Immunohistochemical staining was performed to observe the expression of AQPs in the colon and kidney tissues. Western blot was performed to determine the protein levels of cAMP， PKA， and VIP in the colon tissue. ResultCompared with the control group, the model group had longer time to the first black stool defecation, reduced fecal pellets and water content, reduced Bristol Stool Form Scale score and intestinal propulsion rate, and constipation aggravated（P<0.01）. Moreover, increased the intestinal lesions, reduced the mucus secretion, reduce the serum VIP level, up-regulated the expression levels of AQP1 in the colon and kidney tissues, inhibited the expression of AQP3 and AQP9（P<0.01）., and down-regulated the protein levels of cAMP, PKA, and VIP in the colon tissue. Compared with the model group， the high-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex group had shortened time to the first black stool defecation， increased fecal pellets and water content， increased Bristol Stool Form Scale score and intestinal propulsion rate， and alleviated constipation symptoms. Moreover， high-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex reduced the intestinal lesions， increased the mucus secretion， elevated the serum VIP level（P<0.01）.， down-regulated the expression levels of AQP1 in the colon and kidney tissues， promoted the expression of AQP3 and AQP9（P<0.05，P<0.01）， and up-regulated the protein levels of cAMP， PKA， and VIP in the colon tissue. The medium- and low-dose groups had weaker effect than the high-dose group（P<0.01）. ConclusionHigh-dose Aconiti Lateralis Radix Praeparata-Cinnamomi Cortex can improve the intestinal motility and balance the intestinal water and fluid metabolism by up-regulating the VIP/cAMP/PKA/AQP pathway， thereby mitigating the constipation symptoms in the rat model of slow transit constipation due to yang deficiency.
ABSTRACT
Objective To detect brain structural difference between children with unexplained mental retardation and children with typically normal development. Methods The high-resolution magnetic MR imaging were obtained from 21 children with unexplained mental retardation and 30 age-matched control children without intellectual disabilities. Voxel-based morphometry analysis with an optimization of spatial segmentation and normalization procedures were applied to compare differences of gray matter volume between the two groups. The total and regional gray matter volume were compared between the two groups with independent t test. Meanwhile, correlation was conducted to analyze the relationship between the total gray matter volume and intelligence quotient (IQ) with partial correlation test. Results The total gray matter volume was significantly increased in the mental retardation children [(1. 012 ±0. 079) × 106 mm3]in relative to the controls [(0. 956 ± 0. 059) × 106 mm3, t = - 2. 80, P ＜ 0. 05]. Compared to controls,children with unexplained mental retardation showed significantly increased gray matter volume in different regions, including the bilateral thalami, the bilateral superior frontal gyri, the bilateral gyri rectus, the bilateral temporal poles, the right inferior frontal gyrus, right parahippocampal gyrus and the right cerebellum. No correlation was detected between the total gray matter volume and IQ in children with mental retardation (r = 0. 078 ,P ＞ 0. 05). Conclusions VBM would detect the gray matter abnormalities that were not founded in routine MR scanning. The increase of gray matter volume in the frontal-thalamus network might indicate the delayed maturation of the brain development. This might be one of the causations of mental retardation in children.