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1.
China Pharmacy ; (12): 1825-1832, 2022.
Article in Chinese | WPRIM | ID: wpr-936486

ABSTRACT

OBJECTIVE To explore the the reg ulation of intestinal flora and effects of Qingjie huagong decoction on intestinal mucosal barrier in severe acute pancreatitis (SAP)mode rats . METHODS SAP rat model was induced by intraperitoneal injection of caerulein and lipopolysaccharide.The survival state of rats in each group were observed.The levels of serum amylase ,interleukin 10(IL-10),IL-18 and IL- 1β in serum were all detected. The pathological changes of pancreatic and small intestinal tissue were observed. The expressions of Occludin,ZO-1 and HMGB1 were detected in small intestinal tissue of rats. The structure and relative abundance of intestinal microflora in rats were detected by 16S rRNA high throughput sequencing. RESULTS After the intervention of Qingjie huagong decoction ,abdominal distension symptoms of SAP model rats were significantly relieved ,and their mental state recovered better ;the levels of serum amylase and IL- 18 in serum were decreased significantly (P<0.05),while the level of IL- 10 was increased significantly (P<0.05). The necrotic area of pancreatic tissue and the infiltration of inflammatory cells were reduced , the degree of intestinal epithelial cell structural disorder was alleviated ,and the shedding of intestinal mucosal epithelium was reduced.The protein expression of HMGB 1 in small intestinal tissue was decreased significantly (P<0.05),and the protein expression of Occludin and ZO- 1 were increased significantly . Results of 16S rRNA high throughput sequencing showed that Qingjie huagong decoction could increased the relative abundance of probiotics such as Bacteroidea and Lactobacillus in rat intestine ,reduced the colonization of harmful bacteria such as Firmicutes. CONCLUSIONS Qingjie huagong decoction can improve the intestinal barrier by up-regulating the expression of Occludin and ZO- 1 in small intestinal tissue and down-regulating the protein expression of HMGB 1. It can also adjust the relative abundances of different flora to protect the intestinal tract.

2.
Article in Chinese | WPRIM | ID: wpr-928642

ABSTRACT

OBJECTIVES@#To examine the changes of intestinal flora in children newly diagnosed with acute lymphoblastic leukemia (ALL) and the influence of chemotherapy on intestinal flora.@*METHODS@#Fecal samples were collected from 40 children newly diagnosed with ALL before chemotherapy and at 2 weeks, 1 month, and 2 months after chemotherapy. Ten healthy children served as the control group. 16S rDNA sequencing and analysis were performed to compare the differences in intestinal flora between the ALL and control groups and children with ALL before and after chemotherapy.@*RESULTS@#The ALL group had a significant reduction in the abundance of intestinal flora at 1 and 2 months after chemotherapy, with a significant reduction compared with the control group (P<0.05). Compared with the control group, the ALL group had a significant reduction in the diversity of intestinal flora before and after chemotherapy (P<0.05). At the phylum level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Actinobacteria at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05) and a significant increase in the relative abundance of Proteobacteria at 1 and 2 months after chemotherapy (P<0.05). At the genus level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Bifidobacterium at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05); the relative abundance of Klebsiella in the ALL group was significantly higher than that in the control group at 1 and 2 months after chemotherapy and showed a significant increase at 1 month after chemotherapy (P<0.05); the relative abundance of Faecalibacterium in the ALL group was significantly lower than that in the control group before and after chemotherapy and showed a significant reduction at 2 weeks and 1 month after chemotherapy (P<0.05). The relative abundance of Enterococcus increased significantly at 1 and 2 months after chemotherapy in the ALL group (P<0.05), and was significantly higher than that in the control group (P<0.05).@*CONCLUSIONS@#The diversity of intestinal flora in children with ALL is significantly lower than that in healthy children. Chemotherapy significantly reduces the abundance of intestinal flora and can reduce the abundance of some probiotic bacteria (Bifidobacterium and Faecalibacterium) and increase the abundance of pathogenic bacteria (Klebsiella and Enterococcus) in children with ALL.


Subject(s)
Bacteria/genetics , Bifidobacterium , Child , Feces/microbiology , Gastrointestinal Microbiome , Humans , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy
3.
Article in Chinese | WPRIM | ID: wpr-928640

ABSTRACT

OBJECTIVES@#To study the features of intestinal flora in children with food protein-induced proctocolitis (FPIP) by high-throughput sequencing.@*METHODS@#A total of 31 children, aged <6 months, who experienced FPIP after exclusive breastfeeding and attended the outpatient service of the Third Affiliated Hospital of Zunyi Medical University from October 2018 to February 2021 were enrolled as the FPIP group. Thirty-one healthy infants were enrolled as the control group. Fecal samples were collected to extract DNA for PCR amplification. High-throughput sequencing was used to perform a bioinformatics analysis of 16S rDNA V3-V4 fragments in fecal samples.@*RESULTS@#The diversity analysis of intestinal flora showed that compared with the control group, the FPIP group had a lower Shannon index for diversity (P>0.05) and a significantly higher Chao index for abundance (P<0.01). At the phylum level, the intestinal flora in both groups were composed of Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes. Compared with the control group, the FPIP group had a significant reduction in the composition ratio of Actinobacteria (P<0.001) and a significant increase in the composition ratio of Proteobacteria (P<0.05). At the genus level, the intestinal flora in the FPIP group were mainly composed of Escherichia, Clostridium, Enterococcus, Klebsiella, and Bifidobacterium, and the intestinal flora in the control group were mainly composed of Bifidobacterium and Streptococcus. Compared with the control group, the FPIP group had a significant reduction in the composition ratio of Bifidobacterium and Ruminococcus (P<0.05) and significant increases in the composition ratios of Clostridium and Shigella (P<0.05).@*CONCLUSIONS@#Compared with the control group, the FPIP group has a reduction in the diversity of intestinal flora and an increase in their abundance, and there are certain differences in several bacterial genera. These results suggest that changes in the composition of intestinal flora at genus level may play an important role in the development and progression of FPIP.


Subject(s)
Bacteria/genetics , Bifidobacterium/genetics , Child , Gastrointestinal Microbiome , High-Throughput Nucleotide Sequencing/methods , Humans , Infant , Proctocolitis , RNA, Ribosomal, 16S/genetics
4.
Article in Chinese | WPRIM | ID: wpr-920542

ABSTRACT

Objective To explore the effects of different dietary induction models of inulin, resistant starch RS3 and their complexes on the body weight and intestinal flora in mice. Methods A total of 64 C57BL/6 mice were randomly divided into low-fat control group, low-fat inulin group, low-fat resistant starch RS3 group, low-fat composite group and high-fat control group, high-fat inulin group, high-fat resistant starch RS3 group and high-fat composite group for dietary intervention. The mice were weighed and fresh feces were collected weekly. Diet intervention was continued until the weight of the high-fat control group was more than 14% higher than that of the low-fat control group. The mice were then sacrificed after overnight fasting. Liver and epididymal fat were weighed, and the colon contents were collected for 16S amplicon sequencing analysis. Results In low-fat diet fed mice, the combined induction of inulin and resistant starch RS3 caused a significant decrease in body weight gain. In high-fat diet fed mice, inulin alone and the combined induction both caused a significant reduction in weight gain, and there was no significant difference between the two methods. In the high-fat diet groups, inulin, resistant starch RS3, and the compound could be distinguished by Bacteroides, Bifidobacterium and Alloprevotella respectively. In the low-fat diet groups, inulin, resistant starch RS3, and the composite groups could be distinguished by Coriobacteriaceae_UCG_002, Bacteroides and Helicobacter, respectively. Conclusion Inulin and resistant starch RS3 diet induction can significantly reduce the weight gain of C57BL/6 mice, change the structure of intestinal flora, and show the difference between high-fat and low-fat diets. Inulin and resistant starch RS3 may reduce body weight and promote fat metabolism by changing the structure of intestinal flora.

5.
Acta Pharmaceutica Sinica ; (12): 2580-2589, 2022.
Article in Chinese | WPRIM | ID: wpr-941522

ABSTRACT

As an essential amino acid, tryptophan (Trp) has various physiological functions and is of great significance in the metabolic process of tumors. In the human body, tryptophan is mainly transformed through kynurenine metabolic pathway, which not only promotes the inherent malignant properties of tumor cells, but also leads to immune-suppressive tumor microenvironment. Changes in tryptophan metabolism often occur in tumors, accompanied by abnormal gene expression of tryptophan-related enzymes, among which indoleamine 2,3-bioxygenase (IDO)-related gene expression and tryptophan 2,3-dioxygenase (TDO)-related gene changes are the most significant. A large number of clinical trials on IDO inhibitors, TDO inhibitors and combination therapy have been carried out. This paper reviewed the tryptophan metabolic pathway, regulation of IDO (TDO), kynurenine (KYN) and other related genes in tumor cells, and outlined the development of therapeutic schedule targeting tryptophan-related genes. The new progress provides new ideas for the further exploration of tumor treatment options.

6.
Article in Chinese | WPRIM | ID: wpr-941030

ABSTRACT

OBJECTIVE@#To investigate the effect of Enterococcus faecium QH06 on TNBS-induced ulcerative colitis (UC) in rats and explore the mechanisms in light of intestinal flora and intestinal immunity.@*METHODS@#Thirty-six male Wistar rats were randomized equally into control group, UC model group, and E.faecium QH06 intervention group. The rats in the latter two groups were subjected to colonic enema with 5% TNBS/ethanol to induce UC, followed by treatment with intragastric administration of distilled water or E.faecium QH06 at the dose of 0.21 g/kg. After 14 days of treatment, the rats were examined for colon pathologies with HE staining. The mRNA and protein expression levels of IL-4, IL-10, IL-12, and IFN-γ in the colon tissues were detected using RT-qPCR and ELISA, and the expression of TLR2 protein was detected with immunohistochemistry and ELISA. Illumina Miseq platform was used for sequencing analysis of the intestinal flora of the rats with bioinformatics analysis. The correlations of the parameters of the intestinal flora with the expression levels of TLR2 and cytokines were analyzed.@*RESULTS@#The rats with TNBS- induced UC showed obvious weight loss (P < 0.01) and severe colon tissue injury with high pathological scores (P < 0.01). The protein expression levels of IFN-γ, IL-12, and TLR2 (P < 0.01) and the mRNA expression levels of IFN-γ, IL-12 and IL-10 (P < 0.05) were significantly increased in the colon tissues of the rats with UC. Illumina Miseq sequence analysis showed that in UC rats, the Shannon index (P < 0.05) ACE (P < 0.01)and Chao (P < 0.05) index for the diversity of intestinal flora both decreased with a significantly increased abundance of Enterobacteriaceae (P < 0.01) and a lowered abundance of Burkholderiaceae (P < 0.05). Compared with the UC rats, the rats treated with E. faecium QH06 showed obvious body weight gain (P < 0.05), lessened colon injuries, lowered pathological score of the colon tissue (P < 0.05), decreased protein expressions of IFN- γ, IL- 12, and TLR2 and mRNA expressions of IFN- γ and IL-12 (P < 0.01 or 0.05), and increased protein expressions of IL- 4 (P < 0.05). The Shannon index ACE (P < 0.05) and Chao (P < 0.05) index of intestinal microflora were significantly increased, the abundance of Enterobacteriaceae was lowered and that of Burkholderiaceae and Rikenellaceae was increased in E.faecium QH06- treated rats (P < 0.01 or 0.05). Correlation analysis showed that IFN-γ was positively correlated with the abundance of Enterobacteriaceae, and IFN-γ was negatively correlated with the abundance of Prevotellaceae, Desulfovibrionaceae, norank_o_Mollicutes_RF39 and Clostridiales_vadinBB60_group; TLR2 was negatively correlated with Clostridiales_vadinBB60_group, norank_o_Mollicutes_RF39 and Prevotellaceae.@*CONCLUSION@#E.faecium QH06 can alleviate TNBS-induced colonic mucosal injury in rats, and its effect is mediated possibly by increasing the abundance of SCFA-producing bacteria such as Prevotellaceae and inhibiting abnormal immune responses mediated by TLR2.


Subject(s)
Animals , Colitis, Ulcerative/drug therapy , Colon/metabolism , Interleukin-10 , Interleukin-12/therapeutic use , Male , RNA, Messenger/metabolism , Rats , Rats, Wistar , Toll-Like Receptor 2/metabolism
7.
Article in Chinese | WPRIM | ID: wpr-940786

ABSTRACT

Chronic renal failure (CRF), a common outcome of various chronic kidney diseases, is characterized by retention of metabolites and toxins, water-electrolyte imbalance, acid-base disturbance, and various symptoms in diverse systems. The incidence and progression of this disease are influenced by many factors, particularly the change of intestinal flora. Previous research shows that the intestinal flora interacts with CRF. For CRF patients, the metabolic waste fails to be cleared in time due to the gradual decline of renal function and thus accumulates in vivo. Moreover, CRF changes the composition of intestinal flora, damages intestinal barrier, and accelerates the synthesis of intestinal uremic toxins and the accumulation in blood. As a result, the renal injury is aggravated. The imbalance of intestinal flora can induce acute kidney injury, increase cardiovascular complications, stimulate immune inflammatory responses, and thus aggravate the progression of CRF. Microbiota-targeted therapy for CRF has become the research focus. According to traditional Chinese medicine, kidney disease is related to the intestine and kidney disease should be treated from the intestine. Spleen and kidney are in the closest relationship with the pathogenesis of CRF and the intestinal flora. Chinese medicine, which features multiple targets, multiple effects, and multiple components, acts on the "gut-kidney axis". It is thus superior in the clinical treatment of CRF and the regulation of intestinal flora. To be specific, it intervenes in intestinal flora to delay the process of CRF. In this paper, based on the correlation of traditional Chinese medicine theory with intestinal flora and CRF, this paper reviewed the interaction between intestinal flora and CRF and traditional Chinese medicine intervention in the intestinal flora for the treatment of CRF, which is expected to serve as a reference for the clinical treatment of this disease and the drug development.

8.
Article in Chinese | WPRIM | ID: wpr-940723

ABSTRACT

ObjectiveTo study the effect of Siwutang (SWT) on intestinal flora in rats with diminished ovarian reserve (DOR) induced by Tripterygium wilfordii polyglycoside (TWP) based on 16S rRNA sequencing. MethodTwenty 8-week-old female SD rats were randomly assigned into four groups: blank group, model group, SWT high-dose group, and SWT low-dose group. Except the blank group, the other three groups were underwent intragastric administration of TWP tablets at a dose of 50 mg·kg-1 for 14 days. On day 15, the high-dose group was administrated at 3 times of the human dosage (40 g/person/day), the low-dose group at 1.5 times of the human dosage, and the model group and the blank group with the same volume of normal saline for 18 days. Then, feces were collected for 16S rRNA sequencing. One hour after administration, blood was sampled from abdominal aorta after anesthesia for the measurement of hormone levels by radioimmunoassay, and ovaries were sampled, embedded, sliced, and stained with haematoxylin-eosin (HE) for pathological observation. ResultThe model group had higher level of luteinizing hormone (LH, P<0.05) and lower level of estradiol (E2, P<0.05) than the blank group. The SWT high-dose group and low-dose group had lower LH levels (P<0.05) and higher E2 levels than the model group (P<0.05). SWT reversed the elevation in follicle-stimulating hormone (FSH) and LH levels and the decline in E2 and progesterone (P) levels caused by TWP to some extent. There were a large number of follicles at different developmental stages in the blank group, while atretic follicles increased significantly in the model group. A large number of mature follicles, secondary follicles, and primary follicles were observed in the high-dose SWT group, and primordial follicles, secondary follicles, and increased corpus luteum in the low-dose SWT group. Compared with that in the blank group and the administration group, the abundance of Verrucomicrobia and Epsilonbacteraeota in the model group significantly reduced. Compared with the blank group, the model group had different intestinal flora in phylum, class, order, family, and genus levels. Specifically, the model group had increased proportions of Firmicutes and Bacteroidetes. After TWP modeling, the abundance of Lachnospiraceae decreased significantly while that of Ruminococcaceae UCG-005 increased significantly. SWT groups, blank group, and model group can be clearly distinguished, and SWT groups had a tendency to approach the blank group. ConclusionSWT may improve the ovarian function of rats with TWP-induced DOR by regulating intestinal flora diversity.

9.
Article in Chinese | WPRIM | ID: wpr-940696

ABSTRACT

ObjectiveTo investigate the therapeutic effect and the possible mechanism of Mankuining decoction on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. MethodA total of 90 male SPF C57BL/6 mice were randomly classified into normal group, model group, mesalazine group (0.266 g·kg-1), and high-, and medium-, low-dose (20, 10, 5 g·kg-1) Mankuining decoction groups, with 15 rats in each group. Mice, except the normal group, drank 3% DSS solution for 7 days to induce UC, and administration (ig) started on the day of modeling. The model group and the normal group were given equivalent amount of 0.9% normal saline once a day for 7 days. The general conditions of mice were recorded every day and the disease activity index (DAI) was calculated. On the 8th day, mice were killed by cervical dislocation. All the colons and feces were collected. The length of colon was recorded, and the histopathological changes of colon were observed based on hematoxylin-eosin (HE) staining. The content of inflammatory factors in colon was detected by enzyme-linked immunosorbent assay (ELISA), and the changes of intestinal flora in mouse feces were determined based on 16SrRNA sequencing. ResultCompared with the normal group, the model group had severe colon damage, reduction in colon length (P<0.01), increase in DAI (P<0.01), decrease in interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) in colon(P<0.01), rise of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-17α (IL-17α), and tumor necrosis factor-α (TNF-α) in colon (P<0.05, P<0.01), and decrease in abundance and diversity of intestinal flora. Compared with the model group, mesalazine and high-, medium-, low-dose Mankuining decoction alleviated the colon injury, recovered the length of colon (P<0.01), decreased DAI (P<0.01), increased IL-10 and TGF-β1 in colon (P<0.01), and decreased IL-1β, IL-6, IL-17α, and TNF-α in colon (P<0.01). Moreover, they raised the abundance and diversity of intestinal flora compared with the model group, as manifested by the increase in the abundance of Firmicutes, Akkermansia, Dubosiella, and Blautia and the decrease in the abundance of Bacteroidetes, Muribaculaceae, Clostridia_UCG-014, and Alistipes. ConclusionMankuining decoction has definite effect in treating UC mice, and the effect is positively correlated with the concentration. In addition, different concentration has different influence on the structure of flora. The mechanism is the likelihood that it alleviates the disorder of intestinal flora to restore intestinal immune balance and further promote the recovery of colonic mucosa.

10.
Article in Chinese | WPRIM | ID: wpr-940693

ABSTRACT

ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation (Sham) group, a model (OVX) group, an estrogen (E) group, and an Erxian decoction (EXD) group, with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction, respectively, and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks, the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted, followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group, the OVX group showed reduced serum estrogen level (P<0.01) and increased serum levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05). Compared with the OVX group, the E group and the EXD group showed increased serum estrogen level (P<0.01) and reduced TC and LDL-C (P<0.05). Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera, the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera, and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1, Anaerovibrio, and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes, Firmicutes, Prevotella 9, Ruminococcaceae UCG-014, Ruminococcus 1, and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously, and Erxian decoction could ameliorate the change.

11.
Article in Chinese | WPRIM | ID: wpr-940619

ABSTRACT

ObjectiveTo explore the therapeutic effect and possible mechanism of Banxia Xiexintang and its disassembled prescriptions in regulating the flora disorder induced by mixed antibiotics in young rats. MethodSeventy male BALB/C young rats were randomly assigned into 7 groups: blank group, model group, Bifidobacterium tetralogy viable tablets (0.68 g·kg-1) group, Banxia Xiexintang (9.1 g·kg-1) group, Xinkai (3.19 g·kg-1) group, Kujiang (1.82 g·kg-1) group, and Ganbu (4.1 g·kg-1) group, with 10 rats in each group. Except the blank group, the other groups were given mixed antibiotics by gavage to induce intestinal flora disorder. After 14 days, the rats in different drug groups were administrated with corresponding drugs by gavage, and those in the blank group and model group with the same amount of normal saline once a day for 14 days. After that, fecal samples were collected aseptically for 16S rDNA sequencing of intestinal flora, and lipopolysaccharide (LPS, 10 mg·kg-1) was injected intraperitoneally to induce inflammatory reaction. The tissue morphology of colonic mucosa was observed via hematoxylin-eosin (HE) staining, and the macrophage infiltration of colonic mucosa was observed via toluidine blue staining and immunohistochemistry. The expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) mRNA were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the blank group, the modeling changed the intestinal flora structure of the young rats (P<0.01), damaged the colonic mucosa, reduced the macrophage infiltration, and down-regulated the mRNA levels of IL-1β, IL-6, IL-8, TNF-α, and IL-10 (P<0.01). Compared with the model group, bifidobacterium quadruple viable tablets, Banxia Xiexintang and its disassembled prescriptions increased the diversity of intestinal flora and the relative abundance of beneficial bacteria such as Bacteroidetes and Firmicutes (P<0.01). At the same time, they ameliorated colonic mucosal injury (P<0.05, P<0.01), increased macrophage infiltration (P<0.05, P<0.01), and up-regulated the mRNA levels of IL-6, IL-8, and TNF-α (P<0.01). The mRNA level of IL-1β was up-regulated in Bifidobacterium tetralogy viable tablets, Banxia Xiexintang, Kujiang, and Ganbu groups (P<0.01), and that of IL-10 was up-regulated in Bifidobacterium tetralogy viable tablets, Banxia Xiexintang, Xinkai, and Ganbu groups (P<0.01). ConclusionBanxia Xiexintang and the disassembled prescriptions can adjust the intestinal flora of young rats exposed to antibiotics and protect the immune barrier of colonic mucosa after intestinal flora disorder. In particularly, the whole prescription of Banxia Xiexintang demonstrates the best performance.

12.
Article in Chinese | WPRIM | ID: wpr-940596

ABSTRACT

ObjectiveTo explore the structural characteristics and functional differences of intestinal flora in patients with type 2 diabetes mellitus (T2DM) of dampness heat trapping spleen(DHTS) syndrome and Qi-Yin deficiency(QYD) syndrome. MethodFrom June 2018 to January 2020,62 T2DM patients with DHTS syndrome and 60 with QYD syndrome were selected from Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine. Serum and fecal samples were collected to compare body mass index(BMI),glucose and lipid metabolism,fasting insulin (FINS) and fasting C-peptide (FCP) levels,and homeostasis model assessment of insulin resistance(HOMA-IR) of the two syndrome types. Fecal samples were extracted for DNA database construction,and 16S rDNA high-throughput sequencing was used to analyze and compare the intestinal flora and metabolic pathways. Result① The BMI,fasting plasma glucose(FPG),2-hour postprandial blood glucose (2 h PBG),total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),FINS,FCP,and HOMA-IR were higher in patients with DHTS syndrome than in patients with QYD syndrome,and the high density lipoprotein(HDL) of the former was lower than that of the latter,(P<0.05,P<0.01). ② In terms of species composition and differences,Bacteroidetes, Clostridia and Gammaproteobacteria were dominant at the class level,and the relative abundance of Clostridia,Mollicutes and Verrucomicrobiae in QYD syndrome group was higher than that in DHTS syndrome group. At the order level,Bacteroidales,Clostridiales and Enterobacteriales were mainly found. The relative abundance of Clostridiales,Erysipelotrichales and Verrucomicrobiales in QYD syndrome group was obviously higher than that in DHTS syndrome group,while Aeromonadales in the former was lower than that in the latter (P<0.05). At the family level,Bacteroidaceae,Prevotellaceae and Ruminococcaceae were predominant. The relative abundance of Ruminococcaceae,Porphyromonadaceae and Erysipelotrichaceae in QYD syndrome group was higher than that in DHTS syndrome group(P<0.05). At the genus level,Bacteroides,Prevotella and Parabacteroides were mainly found. The relative abundance of Parabacteroides,Butyrivibrio and Ruminiclostridium in QYD syndrome group was higher than that in DHTS syndrome group,while that of Klebsiella and Megasphaera in DHTS syndrome group was higher than that in QYD syndrome group(P<0.05). ③ Through Venn analysis of operational taxonomic units(OTU),it was found that there were 49 OTUs in patients with DHTS syndrome patients and 47 OTUs in QYD syndrome patients. ④ The results of OTU β diversity and α analysis showed that Shannon and Simpson indexes had statistical differences,while Ace and Chao indexes had no statistical differences. The intestinal microbial diversity of patients with QYD syndrome was higher than that of patients with DHTS syndrome(P<0.05). The analysis of similarities (ANOSIM) showed that the difference of β diversity between the two groups was significant(P<0.05). ⑤ Linear discriminant analysis Effect Size(LEfSe) results demonstrated that Klebsiella,Megasphaera and Aeromonadales could be selected as the key biomarkers for DHTS syndrome; 14 bacteria such as Ruminiclostridium,Burkholderiaceae,Lautropia,Butyrivibrio,Erysipelotrichales can be selected as the key biomarkers for QYD syndrome. ⑥Functional annotation and analysis showed that the DHTS syndrome involved 9 metabolic pathways,including arginine and proline metabolism,lipopolysaccharide biosynthesis,nicotinic acid and nicotinamide metabolism,while the QYD syndrome involved 10 metabolic pathways,including acarbose and valinomycin biosynthesis,glucagon signaling pathway and NOD-like receptor signaling pathway. ConclusionThere are obvious differences in intestinal flora and functions in T2DM patients of DHTS syndrome and QYD syndrome,which can be used as reference for traditional Chinese medicine (TCM) syndrome differentiation and the target of TCM treatment.

13.
Article in Chinese | WPRIM | ID: wpr-940491

ABSTRACT

ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout (ApoE-/-) mice, and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group, rosuvastatin group (10 mg·kg-1), high-, low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis (75, 25 mg·kg-1), with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration, blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin (HE) staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group, the plaque area of the model group was significantly increased with inflammatory infiltration, the contents of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), inflammatory factors and inducible nitric oxide synthase (iNOS) were increased, while the content of high-density lipoprotein cholesterol (HDL-C) was decreased. Compared with the model group, rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque, reduce the contents of TG, TC, LDL-C, inflammatory factors and iNOS, and increase the content of HDL-C. Compared with the blank group, the relative abundances of Firmicutes and Proteobacteria in the model group increased, while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated, and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group, ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids, which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors, improve antioxidant capacity and repair the disorder of intestinal flora structure.

14.
Article in Chinese | WPRIM | ID: wpr-940481

ABSTRACT

ObjectiveTo investigate the effect of Shaoyaotang on diarrhea, inflammation, and intestinal flora in rats with dampness-heat diarrhea and explore the mechanism of therapeutic principle "treating incontinent syndrome with dredging method" of Shaoyaotang. MethodThe dampness-heat diarrhea model was induced by high temperature, high humidity, high sugar and fat diet, and pathogenic factors. The rats were divided into normal group, model group (normal saline), Shaoyaotang group (5.62 g·kg-1), Rhei Radix et Rhizoma (RRER)-free Shaoyaotang group (5.15 g·kg-1), and RRER group (0.01 g·kg-1). The rats were treated correspondingly for five days, twice a day in the morning and evening. The diarrhea index was used to evaluate the antidiarrheal effect of each group three hours after the administration in the evening. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-2, and IL-6 in the serum were detected by enzyme-linked immunosorbent assay (ELISA) three hours after the last administration. The structure of intestinal flora in feces was characterized by 16sDNA. ResultCompared with the model group, the Shaoyaotang group, the RRER-free Shaoyaotang group, and the RRER group showed reduced diarrhea index (P<0.01), with the onset rates ranking as the Shaoyaotang group>the RRER-free Shaoyaotang group>the RRER group. Those three groups with drug intervention all showed decreased levels of inflammatory factors (P<0.01), especially the Shaoyaotang group, and no significant difference was observed between the RRER group and the RRER-free Shaoyaotang group. The abundance of pathogenic bacteria and conditioned pathogens (e.g. Escherichia-Shigella, Prevotella, Enterorhabdus, and Bacteroides) was reduced and the proliferation of probiotics (such as Ruminococcus, Turicibacter, and Lachnospiraceae) was increased in the groups with drug intervention (P<0.01). For the structure of intestinal flora, the RRER group and the Shaoyaotang group were close to the normal group, and the RRER-free Shaoyaotang group was different from the other three groups (P<0.01). ConclusionShaoyaotang can improve the outcome of rats with dampness-heat diarrhea through anti-inflammation and regulation of intestinal flora disorders. RRER in the prescription plays a key role in reducing the abundance of harmful bacteria and promoting the proliferation of probiotics, which is the key of Shaoyaotang in promoting the re-balance of intestinal flora. It also confirms the scientificity of treating dampness-heat diarrhea with RRER following the therapeutic principle "treating incontinent syndrome with dredging method".

15.
Article in Chinese | WPRIM | ID: wpr-940459

ABSTRACT

ObjectiveTo investigate the effect of Shunao Jieyu decoction on intestinal flora in patients with post-stroke depression. MethodSixty patients with post-stroke depression of Qi stagnation, blood stasis, and phlegm obstruction were selected and divided into a treatment group (30 cases, Shunao Jieyu decoction) and a control group (n=30, paroxetine hydrochloride tablets) according to the random number table. All patients were treated correspondingly for eight weeks. The scores of traditional Chinese medicine(TCM) syndrome, Hamilton rating scale for depression(HAMD), National Institutes of Health stroke scale(NIHSS), and activities of daily living(ADL)before and after treatment were compared between the two groups. High-throughput sequencing was used to analyze the diversity of fecal flora and the distribution of taxonomical levels in two groups before and after treatment. ResultThe post-treatment TCM syndrome score, HAMD score, and NIHSS score were lower than those before treatment in the same group (P<0.05), while the post-treatment ADL score was higher than that before treatment (P<0.05). Compared with the control group after treatment, the treatment group showed decreased TCM syndrome score (P<0.05). No significant difference was observed in the HAMD score, NIHSS score and ADL score between the two groups after treatment. The total effective rate of the treatment group was 90% (27/30), which was superior to 66.3% (19/30) of the control group (χ2=5.863, P<0.05). After treatment, the average values of Chao1 index, Observed species index, Shannon index, Simpson index, and Pielou's evenness index of intestinal flora diversity in the treatment group increased without significant difference, while the average value of the Good's Coverage index remained unchanged in the same group. At the phylum level, the abundance of Bacteroidetes increased. At the family level, the abundance of Bacteroidaceae increased. At the genus level, the abundance of Bacteroidetes increased. ConclusionShunao Jieyu decoction can effectively improve the clinical TCM symptoms of patients with post-stroke depression, relieve neurological impairment, improve the ability of daily living, and change the diversity and abundance of the intestinal flora of patients at different taxonomic levels.

16.
Article in Chinese | WPRIM | ID: wpr-940444

ABSTRACT

Intestinal flora is the largest microbial community in human body, which consists of more than 1 000 species. Its structure and metabolites change dynamically with the age, diet and intestinal environment of the host. Study shows that the intestinal microbes play a pivotal role in regulating human physiological and pathological processes, and intestinal flora imbalance may be the key factors affecting the occurrence and development of bone and joint diseases, including osteoporosis, osteoarthritis, rheumatoid arthritis and gouty arthritis. At present, calcitonin, estrogen, nonsteroidal anti-inflammatory drugs, immunosuppressants, xanthine oxidase inhibitors and other western drugs are mostly used to treat the above diseases. However, long-term use of western drugs leads to poor compliance and obvious gastrointestinal adverse reactions among patients. Traditional Chinese medicine (TCM) predominates in the treatment of bone and joint diseases due to its low price, high efficacy and slight side effects, with the advantages of multi-targets, multi-mechanism and multi-levels. In recent years, many scholars have carried out experiments and clinical studies on the treatment of bone and joint diseases by TCMs on the basis of the liver and kidney theory such as "tonifying liver and kidney and strengthening muscles and bones". Gratifying results have been achieved. However, the mechanism of action has not been fully clarified. Intestinal flora becomes a hot spot in medical research, and a close relationship between intestinal flora and bone and joint diseases has been unveiled. Relevant literature in China and abroad showed that TCM has a significant effect on the treatment of bone and joint diseases by regulating intestinal flora. In this paper, the relationship between intestinal flora and bone and joint diseases was summarized and the intervention of TCM active ingredients and compounds on intestinal flora was reviewed to facilitate the prevention and treatment of bone and joint diseases by TCM.

17.
Article in Chinese | WPRIM | ID: wpr-940429

ABSTRACT

ObjectiveTo observe the clinical efficacy of Maxingshigantang enema in the treatment of infant viral pneumonia by comparing related indicators, and comprehensively evaluate the effect of traditional Chinese medicine (TCM) enema on the intestinal microenvironment. MethodSixty infants with viral pneumonia were selected and randomly divided into 3 groups. The dosage of enema drugs in high- (0.117 g·mL-1) and low-concentration (0.07 g·mL-1) TCM enema groups was same (3.5 g per time), and the control group received normal saline enema, once a day for 7 days. Finally, the curative effect, total symptom score, salivary secretory immunoglobulin A (sIgA), human beta defensin 2 (hBD2) and fecal calprotectin (CALP) of each group were statistically analyzed by SPSS 21.0, and the clinical efficacy of TCM enema in treating children with pneumonia and asthma was comprehensively evaluated. ResultThe curative effect of high-concentration TCM enema group (total effective rate 100%, χ2=7.059) was equivalent to that of low-concentration TCM enema group (total effective rate 95%, χ2=4.329), higher than that of control group (total effective rate 70%) (P<0.017). After treatment, compared with control group and low-concentration TCM enema group, high-concentration TCM enema group had higher total symptom score of children (P<0.05, P<0.01). The proportion of coccobacillus was reduced in three groups, with high- and low-concentration TCM enema groups lower than control group (P<0.05). The salivary sIgA concentration was increased in three groups (P<0.05), with high-concentration TCM enema group higher than the other groups (P<0.01). The hBD2 concentration was decreased in three groups, with high- and low-concentration TCM enema groups lower than control group (P<0.05). The three groups reduced the fecal CALP concentration, and high-concentration TCM enema group had the highest reduction, followed by low-concentration TCM enema group (P<0.01). ConclusionTCM enema outweighs western medicine in improving clinical symptoms, intestinal flora, and mucosal immune function, and reducing inflammation in children, and the high-concentration TCM enema group has better curative effect. Therefore, with easiness to operate, high compliance, and significant therapeutic effect, TCM enema is worthy of clinical promotion.

18.
Article in Chinese | WPRIM | ID: wpr-940409

ABSTRACT

Alzheimer's disease (AD) is a neurodegenerative disease. With the acceleration of aging process, the number of AD patients increases year by year. This threatens the health and even life of patients, and causes heavy economic burden and mental pressure to patients, families and society. In traditional Chinese medicine (TCM), AD belongs to the category of dementia, and tonifying kidney is the main treatment. Based on the basic theory of TCM and combined with clinical manifestations of AD, AD is closely correlated with liver and spleen. Therefore, "simultaneous regulation of three Yin" of liver, spleen and kidney will be an important way for the prevention and treatment of AD. Hei Xiaoyaosan, a representative prescription of "simultaneous regulation of three Yin" of liver, spleen and kidney, has theoretical, experimental and clinical basis in preventing and treating AD. Modern studies have shown that neurofibrillary tangle formed by tau hyperphosphorylation is a main pathological feature of AD, and trimethylamine oxide (TMAO) is closely related to tau hyperphosphorylation. Therefore, regulating TMAO metabolism to inhibit tau hyperphosphorylation is a new target for the prevention and treatment of AD. On the basis of the above theory and previous studies, this paper put forward the hypothesis that Hei Xiaoyaosan regulates the trimethylamine(TMA)/heparin monooxygenase 3(FMO3)/TMAO metabolic pathway of intestinal flora through "simultaneous regulation of three Yin" of liver, spleen and kidney, and then inhibits tau hyperphosphorylation in brain hippocampus, thereby protecting nerve cells, improving learning and memory, and preventing AD. This paper explored the role and mechanism of Hei Xiaoyaosan in the prevention and treatment of AD from the perspective of inhibiting tau hyperphosphorylation by regulating the TMA/FMO3/TMAO metabolic pathway of intestinal flora, which provided new ideas and strategies for in-depth study of Hei Xiaoyaosan in the prevention and treatment of AD.

19.
Article in Chinese | WPRIM | ID: wpr-940296

ABSTRACT

ObjectiveTo study the effect of Huazhuo Jiedu Huoxue Tongluo (HJHT) prescription on the intestinal flora in rats with cerebral ischemia-reperfusion injury, and to explore the mechanism of Chinese medicinal prescription regulating intestinal flora to restore the balance of brain-gut axis. MethodFifty male SPF SD rats were randomly assigned into sham group, model group, high-dose HJHT group (25.0 g·kg-1), medium-dose HJHT group (12.5 g·kg-1), and low-dose HJHT group (6.25 g·kg-1), with 10 rats in each group. The rat model of permanent middle cerebral artery infarction was established according to Longa method and previous research experience, and reperfusion was performed 2 h after ischemia. The recovery of neurological function deficit and the percentage of cerebral infarction area were detected 72 h after administration. Real-time PCR was performed to detect the mRNA levels of Occludin and zonula occludens-1 (ZO-1) in rat colon. Hematoxylin-eosin (HE) staining was conducted to reveal the intestinal damage. The feces of 6 rats in each group were collected for 16S rRNA sequencing. The expression of Treg and Th17 in intestinal tissue, peripheral blood, and brain tissue were detected. ResultCompared with the sham group, the model group showed obvious neurological deficit (P<0.05) and large cerebral infarction area (P<0.05). High-dose and medium-doses HJHT alleviated the symptoms of neurological impairment (P<0.05) and reduce the cerebral infarction area (P<0.05) compared with the model group. Compared with the sham group, the model group showed destroyed structure of colonic mucosa and incomplete epithelial cells and goblet cells, while high-dose and medium-doses HJHT alleviated such changes. The mRNA levels of Occludin and ZO-1 in the model group were lower than those in the sham group (P<0.05),and the high-dose HJHT groups were higher than the model group (P<0.05). The intestinal flora structure was significantly different between the model group and the sham group while similar between the high-dose HJHT group and sham group. Compared with the sham group, the model group showed down-regulated expression of Treg and up-regulated expression of Th17 in the intestinal tissue, peripheral blood, and brain tissue, and high-dose and medium-dose HJHT alleviated the changes in the expression of Treg and Th17 in the model group (P<0.05). ConclusionHuazhuo Jiedu Huoxue Tongluo prescription may improve the permeability of intestinal wall by adjusting the abundance and diversity of intestinal microorganisms to reduce the migration of intestinal Th17 cells toward the ischemic lateral brain tissue, mitigate the inflammatory response, and thus alleviate the cerebral ischemia-reperfusion injury in rats.

20.
Article in Chinese | WPRIM | ID: wpr-940228

ABSTRACT

With the gradual aggravation of aging in China, the prevalence of osteoporosis is increasing year by year. Osteoporosis has become a major public health problem threatening the health of middle-aged and elderly people, especially middle-aged and elderly women. There are many predisposing factors and complex pathogenesis of osteoporosis. The interpretation of osteoporosis has been the focus of clinical research in recent years. How to prevent and treat osteoporosis more effectively has also become a major problem faced by researchers. In recent years, the balance and homeostasis of calcium and phosphorus regulated by intestinal absorption, renal excretion and bone have become one of the hot topics, and the balance and homeostasis of calcium and phosphorus in vivo are the key to normal bone homeostasis. At the same time, as a complex microbial community living in the gastrointestinal tract, intestinal flora can produce a variety of regulators affecting metabolism. It has been widely confirmed that it acts on the body indirectly or directly, in multiple ways and targets to prevent and treat osteoporosis. Therefore, further exploring the role and mechanism of intestine kidney bone axis in osteoporosis plays a far-reaching significance for the prevention and treatment of osteoporosis. In recent years, scholars have made a lot of exploration on the prevention and treatment of osteoporosis with traditional Chinese medicine (TCM), and found that TCM can intervene the expression of intestinal flora and play the effect of prevention and treatment of osteoporosis. Based on the "intestine kidney bone axis", this paper briefly discusses the integrated traditional Chinese and western medicine of kidney and osteoporosis, intestine and osteoporosis, intestine kidney axis, the treatment of kidney from intestine, intestine and osteoporosis, and the application of TCM in regulating intestinal flora in osteoporosis, in order to provide new ideas for the prevention and treatment of osteoporosis.

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