Objective To explore the protective effect of hydroalcoholic extract of Portulaca oleracea L.(POHA)on ulcerative colitis(UC)and bone loss in mice.Methods The C57BL/6 mice were treated with dextran sulfate sodium(DSS)to establish UC model.A total of 50 mice were randomly assigned to including control group,DSS group,mesalazine(MS)group,low dose of POHA(POHAL)group,or high dose of POHA(POHAH)group.The control group freely drank drinking water,while the DSS,MS,POHAL and POHAH groups drank drinking water containing DSS for 8 weeks.Since the 2nd week,the control group and DSS group were given normal saline by gavage.The MS group was given MS(100 mg/kg)by gavage.The POHAL group and POHAH group were given POHA(1 000 mg/kg and 2 000 mg/kg)by gavage,respectively.Body weight and disease activity index(DAI)were recorded and calculated every 2 d.On the 56th day,the colon weight index,liver index,and spleen index were calculated,and the histological changes of colon were observed.Serum levels of bone metabolism markers and microstructure parameters of femur were detected.Results Compared with the control group,the DSS group showed significantly increased DAI score,colon weight index,liver index,and spleen index(all P＜0.01).The DSS group exhibited significant pathological damage in colon tissues and significantly increased serum levels of osteocalcin,C-terminal peptide of collagen type Ⅰ,and tartrate-resistant acid phosphatase 5b(P＜0.01).The bone loss was significant in the DSS group,manifested by markedly decreased bone mineral density(BMD),bone tissue volume to tissue volume ratio(BV/TV),trabecular bone number(Tb.N),and trabecular bone thickness(Tb.Th),and markedly increased bone surface to bone volume ratio(BS/BV)and trabecular bone separation(Tb.Sp)(P＜0.05 or P＜0.01).Compared with the DSS group,the BMD,BV/TV,Tb.N and Tb.Th of the femur in the MS group and POHAH group of mice were all increased(P＜0.05 or P＜0.01),the BS/BV all decreased(P＜0.05 or P＜0.01),and the Tb.Sp all decreased without significant differences(all P＞0.05).The above bone microstructure parameters in the POHAL group showed no significant differences compared with those in the DSS group(all P＞0.05).Conclusion POHA has protective effect on DSS-induced UC and bone loss,and the mechanism may be related to the inhibition of hyperactive bone metabolism.

A new rapid, quality control method based on quantitative water tests has been established for the quality evaluation of Indigo Naturalis. The Turbiscan stability index (TSI) of 26 batches of Indigo Naturalis was measured by a stability analyzer. The parameters, including the method by which the ingredients are added, their particle size, amount, and the testing temperature, were systematically optimized and the methodological indexes such as repeatability and stability were determined. The content of indigo and indirubin in 26 batches of Indigo Naturalis was determined by high performance liquid chromatography and the total ash was measured. The correlation analysis between the active ingredients, total ash content and TSI value of Indigo Naturalis was determined by SPSS 26.0 and Origin 2021. This research shows that the best way to prepare samples for testing is to add 0.2 g of Indigo Naturalis powder which has passed through a No. 7 sieve but failed to pass through a No. 9 sieve to a glass bottle containing 20 mL pure water by a funnel and scan at 25 ℃ with a stability analyzer. Consistency analysis showed that the content ranking of indigo and indirubin is opposite to the TSI value, and the content ranking of total ash is generally consistent with the TSI value. Correlation analysis showed that the correlation coefficients of indigo and indirubin content and TSI value were -0.850 and -0.801, respectively, and R² was 0.72 and 0.64. The correlation coefficient between total ash content and TSI value was 0.724, R² was 0.52. Using the change in TSI value of Indigo Naturalis powder in water, this study establishes the range of classification of Indigo Naturalis decoction pieces and the content of relevant components, which can be used to authenticate Indigo Naturalis and evaluate its quality.

The soaking and fermentation of Baphicacanthus cusia( Nees),the important intermediate link of Indigo Naturalis processing,facilitates the synthesis of indigo and indirubin precursors and the dissolution of endogenous enzymes and other effective components,while the role of microorganisms in the fermentation is ignored. The present study investigated the changes of microbial community structure in Indigo Naturalis processing based on 16 S amplicon sequencing and bioinformatics. Meanwhile,the contents of indigo,indirubin,isatin,tryptanthrin,indole glycoside,etc. were determined to explore the correlation between the microorganisms and the alterations of the main components. As demonstrated by the results,the microbial diversity decreased gradually with the fermentation,which bottomed out after the addition of lime. Proteobacteria,Bacteroidetes,and Firmicutes were the main dominant communities in the fermentation. The relative abundance of Proteobacteria declined gradually with the prolongation of fermentation time,and to the lowest level after the addition of lime. The relative abundance of Firmicutes increased,and that of Bacteroidetes decreased first and then increased. The contents of effective substances in Indigo Naturalis also showed different variation tendencies. As fermentation went on,indole glycoside decreased gradually; indigo first increased and then decreased; indirubin and isatin first decreased and then increased; tryptanthrin gradually increased. Those changes were presumedly related to the roles of microorganisms in the synthesis of different components. This study preliminarily clarified the important role of microorganisms in the soaking and fermentation and provided a scientific basis for the control of Indigo Naturalis processing and the preparation of high-quality Indigo Naturalis.
Fermentation ; Indigo Carmine ; Indigofera ; Indoles ; Microbiota

As an effective antipyretic medicine,Indigo Naturalis has a long history of application in the field of Chinese medicine.The content of organics,mainly indigo and indirubin,is about 10%. However,the active ingredients and mechanism of its antipyretic effect have not yet been fully elucidated. In view of this,they were investigated in this study with the rectal temperature change as an indicator and 2,4-dinitrophenol-induced fever rats as subjects. The content of PGE2 and c AMP in the hypothalamus and the serum levels of TNF-α,IL-1β and IL-6 were determined by ELISA. Moreover,the plasma samples of fever rats were analyzed by metabonomics in combination with UPLC-Q-TOF-MS for the exploration of potential biomarkers and the discussion on the antipyretic mechanism of Indigo Naturalis and its active ingredients. The results showed that the rising trend of rectal temperature in rats was suppressed 0. 5 h after the treatment with Indigo Naturalis,organic matter,indigo or indirubin as compared with the rats of model group( P < 0. 05),among which Indigo Naturalis and organic matter had better antipyretic effect. ELISA results showed that organic matter and indigo can inhibit the expression of PGE2 and c AMP( P<0. 01),while Indigo Naturalis and organic matter were effective in curbing the increase in TNF-α( P<0. 05). A total of 21 endogenous metabolites were identified from the plasma samples of the Indigo Naturalis,organic matter,indigo and indirubin groups,which were mainly involved in glycerophospholipid metabolism.
2,4-Dinitrophenol ; Animals ; Antipyretics ; Drugs, Chinese Herbal ; Indigo Carmine ; Indigofera ; Rats

Objective: To investigate the effect of Coptidis Rhizoma on intestinal microflora of normal and type 2 diabetic rats before and after being steamed with rice wine based on 16S rRNA sequencing technology. Method: The type 2 diabetic rat model was established by high-fat diet and streptozotocin. Normal and model rats were given the decoction of Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine every day (0.8 g·kg^-1, according to raw drug quantity), the positive drug group was given metformin solution every day (0.25 g·kg^-1), continuous administration for 30 d. Blood glucose was measured once a week and oral glucose tolerance was measured on the 27^th day of administration. On the 30^th day, feces were taken and the rats were anesthetized. Blood was taken for enzyme linked immunosorbent assay (ELISA). Colon was dissected for hematoxylin-eosin (HE) staining. Based on 16S rRNA gene sequencing technology, the alpha diversity, beta diversity, abundance and composition, and the correlation with anti-inflammatory and antioxidant activities of intestinal microflora were comprehensively analyzed and evaluated. Result: After giving normal rats Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine, there were some inflammatory reactions, imbalance of oxidation-antioxidant system, pathological damage of colon and changes of intestinal microflora, which showed some toxic and side effects, but the toxic and side effects of Coptidis Rhizoma steamed with rice wine were lower than those of Coptidis Rhizoma. Coptidis Rhizoma could reduce fasting blood glucose and improve glucose tolerance in type 2 diabetic rats before and after steaming. Although the effect of Coptidis Rhizoma steamed with rice wine had an increasing trend, there was no significant difference between before and after processing. Type 2 diabetic rats appeared mild inflammation and imbalance of oxidation-antioxidant system, characterized by significant increase of contents of interleukin (IL)-6, nuclear transcription factor (NF)-κB and malondialdehyde (MDA) (P<0.05, P<0.01), significant decrease of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) activities (P<0.01). Coptidis Rhizoma could inhibit inflammation and imbalance of oxidation-antioxidant system, and alleviate pathological damage of colon before and after steaming in type 2 diabetic tats, the effect of Coptidis Rhizoma steamed with rice wine was better than that of Coptidis Rhizoma. Significant intestinal flora disorders were observed in type 2 diabetic rats, including down-regulation of Bacteroidetes, Firmicutes and Kiritimatiellaeota, up-regulation of Proteobacteria, Actinobacteria, Acidobacteria, Chloroflexi, Cyanobacteria and Verrucomicrobia. Coptidis Rhizoma could alleviate intestinal microflora disturbance before and after steaming, and the regulation effect of Coptidis Rhizoma on Kiritimatiellaeota, Cyanobacteria and Verrucomicrobia was stronger than that of Coptidis Rhizoma steamed with rice wine. Coptidis Rhizoma steamed with rice wine could significantly reduce the level of Chloroflexi, but Coptidis Rhizoma had no such effect. The intestinal microflora may be correlated with the antioxidant capacity of Coptidis Rhizoma before and after steaming. Conclusion: Coptidis Rhizoma can treat type 2 diabetic rats by regulating intestinal microflora before and after steaming with rice wine, and the effect of Coptidis Rhizoma steamed with rice wine is better. The toxic and side effects of Coptidis Rhizoma on intestinal flora are reduced by steamed with rice wine. The processing mechanism of increasing efficacy and decreasing toxicity of Coptidis Rhizoma steamed with rice wine is closely related to intestinal microflora.