Anti-inflammatory effect, plasma effective components and therapeutic targets of Huanglian Jiedu Decoction on ulcerative colitis mice / 中国中药杂志

This paper was to investigate the effect of Huanglian Jiedu Decoction(HLJD) on ulcerative colitis(UC) in mice, and determine the effective components in plasma, and virtually screen its therapeutic target, and predict its mechanism. Sixty Balb/c mice were randomly divided into blank group, model group, mesalazine treatment group(0.3 g·kg~(-1)), and HLJD treatment groups(24.66, 12.33, 6.17 g·kg~(-1)). Excepted for the blank group, all the mice in HLJD and mesalazine treatment groups were gavage administration. All mice freely drank 2.5% DSS solution for seven days to induce UC. The disease activity index(DAI) was detected each day. At the end of the experiment, HE staining was used to observe the pathological changes in colon. The content of IL-1β, IL-6 and TNF-α in colon were determined by ELISA. The effective components in plasma were determined by UPLC-Q-TOF-MS. The reverse docking in PharmMapper was used to screen the component targets. The disease targets of UC were collected by searching TTD, OMIM and GeneCards databases. The intersection of the component targets and disease targets was selected as the therapeutic targets. Then the therapeutic targets were imported into the STRING for GO and KEGG enrichment analysis. Discovery Studio was used to simulate the docking between the components and the targets. RESULTS:: showed that the DAI in the model group increased significantly(P<0.05), and the number of inflammatory cells and infiltration degree increased significantly compared with the blank group. The DAI in HLJD treatment group was significantly reduced(P<0.05), and the number and infiltration degree of inflammatory cells were reduced compared with the model group. The ELISA results showed that the levels of IL-1β, IL-6 and TNF-α were increased significantly in the model group(P<0.01) compared with the blank group, and significantly down regulated in the HLJD treatment group(P<0.05) compared with the model group. After UPLC-Q-TOF-MS analyse, ten components were identified. The network pharmacology analysis showed that the action targets were significantly enriched in 129 of biological processes, such as response to organic substance, chemical and oxygen-containing compound, etc., as well as 16 of signal pathways, such as IL-17, TNF and hepatitis B signal pathways, were enriched too. The results of molecular docking showed that limonin, palmatine and berberine could bind to CASP3 and MMP9 by hydrogen bond. In conclusion, HLJD could alleviate the colonic mucosal inflammatory infiltration and mucosal damage in UC mice. The mechanism may be related to the anti-inflammatory effect on UC mice by reducing the levels of IL-1β, IL-6 and TNF-α in colon through limonin, palmatine and berberine regulating IL-17 signal pathway and TNF signal pathway via CASP3 and MMP9 meditated.

Effect and mechanism of total flavonoids of Lichi Semen on CCl_4-induced liver fibrosis in rats, and prediction of Q-marker / 中国中药杂志

This paper was to investigate the effect of total flavonoids of Lichi Semen(TFL) on carbon tetrachloride(CCl_4)-induced liver fibrosis in rats, analyze and predict its mechanism of action and potential quality markers(Q-marker). Firstly, male SD rats were taken and injected subcutaneously with a 40% CCl_4-vegetable oil solution twice a week for 8 consecutive weeks to establish a rat model of liver fibrosis. The rats with liver fibrosis were randomly divided into model group, silybin group(43.19 mg·kg~(-1)), Fuzheng Huayu Capsules group(462.75 mg·kg~(-1)), and TFL groups(100 mg·kg~(-1) and 25 mg·kg~(-1)), with normal rats as a blank group, 10 rats in each group. Except for the blank group, the rats in the other groups were subcutaneously injected with 40% CCl_4-vegetable oil solution of a maintenance dose, once a week. The rats in various treatment groups received corresponding doses of drugs, while the rats in the blank group and model group received the same volume of normal saline once a day for 4 weeks. At the end of the experiment, blood was collected from the abdominal aorta and the liver tissues were collected. The levels of total bilirubin(TBiL), direct bilirubin(DBiL), indirect bilirubin(IBiL), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) in serum were detected by using an automatic biochemical detector. Masson staining was used to observe the histopathological changes of rat liver. Then, the chemical compositions of TFL were collected, and the action targets of these chemical compositions were predicted through SWISS database and reverse molecular docking server(DRAR-CPI). After screening of disease targets of liver fibrosis by Gene Cards database, the protein-protein interaction was analyzed with use of STRING database, and GO(gene ontology) analysis and KEGG(Kyoto encyclopedia of genes and genomes) enrich analysis were also carried out. Moreover, an iTRAQ proteomics technology was used to determine protein expression in liver tissues of rats in TFL, model and blank groups to verify the targets. Furthermore, Cytoscape software was used to establish and visualize the network of chemical components, targets and pathways, and predict the potential Q-marker of TFL. The results showed that the levels of TBiL, DBiL, IBiL, ALT, and AST in the model group were significantly higher than those in the blank normal group(P<0.05), and the above levels in the treatment groups were lower than those in the model group, but with no significant differences. Masson staining showed that the liver damage and the degree of fibrosis were severe in the model group, and were relieved to different degrees in the treatment groups. Then, 74 chemical components were screened, which could act on 865 targets such as EGFR and SRC, participating in the regulation of cancer pathways, PI3 K-Akt signaling pathway, HIF-1 signaling pathway and other signaling pathways closely related to liver fibrosis. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin showed the highest correlation with liver fibrosis-related targets and pathways. Proteomics results showed that a total of 18 proteins among the 45 proteins predicted by internet pharmacology were identified, among which 6 proteins were significantly expressed, including 5 up-regulated proteins and 1 down-regulated protein. The protein expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1 was significantly returned to a normal state in the TFL treatment groups. In conclusion, TFL may demonstrate the anti-hepatic fibrosis and potential hepatoprotective effects by regulating the expression of ALB, PLG, HSP90 AA1, EGFR and MAP2 K1, which may be associated with the regulation of multiple signaling pathways related to liver fibrosis such as PI3 K-Akt pathway. Pinocembrin, quercetin, epicatechin, procyanidin A2, naringenin, nobiletin, phlorizin and rutin could be regarded as potential Q-markers of TFL for quality control.

Effect and mechanism of YAO medicine compound containing Cissus pteroclada on rheumatoid arthritis in rats and its Q-marker prediction / 中草药

Objective: To evaluate the anti-rheumatism effect, and predict the mechanism and Q-marker of YAO medicine compound containing Cissus pteroclada (Sifangteng in Chinese, SFT) in the treatment of rheumatoid arthritis (RA) in rats. Methods: SD rats were randomly divided into control group, model group, tripterygium glycosides group and SFT high/low dose (28.7, 7.2 g/kg) groups with eight rats in each group. Except the control group, the RA models in rats induced by Collange II collagen were established. The SFT group and the tripterygium glycosides group were given corresponding drugs by intragastric administration during the modeling period, while the other two groups were given the same volume of saline once daily for 28 d. The degree of foot swelling was measured and scored during the experiment. The levels of TNF-α and IL-1β in serum were measured by ELISA at the end of the experiment. SFT chemical components and predicting targets were searched and screened through TCMSP and Drugbank databases. The target of RA disease was searched by TTD database. The protein interaction network was constructed and visualized by String database and Cytoscape software, cluster analysis was analyzed by MCODE. GO and KEGG enrichment analysis was carried out using String database. Finally, combined with the validity and measurability of chemical components, the Q-marker of SFT was predicted. Results: Compared with the model group, the foot swelling of rats in SFT high and low dose groups and positive group was significantly reduced (P < 0.01), and the serum levels of IL-1β and TNF-α were significantly decreased (P < 0.01). There were 89 disease targets of RA. The pathogenesis of RA was related to abnormal cytokine-receptor pathway and RA pathway. A total of 31 components in SFT were screened and its 119 target proteins were predicted, 12 of them belong to disease targets were involved in 1 112 biological processes, such as regulation of stimulation response, regulation of cell proliferation, regulation of cell metabolism, regulation of intracellular signal transduction, and regulation of 113 signaling pathways, such as RA pathway and TNF pathway, which ultimately play a role in the treatment of RA. At the same time, 11 components were predicted to be Q-markers of SFT, including apigenin, resveratrol, bergenin, nitidine, osthol, linalool, ammidin, ethoxychelerythrine, coptisine, hesperidin, and sesamin. Conclusion: SFT can significantly reduce acute inflammation in RA rats. SFT may act on PTGS2-based targets through resveratrol and other components, and participate in regulation of RA pathway, TNF pathway and other inflammatory and immune pathways. Apigenin, resveratrol and bergenin, nitidine, osthol, linalool, ammidin, ethoxychelerythrine, coptisine, hesperidin, sesamin can be used as Q-markers for SFT quality control.

THE RESEARCH OF FOUR DRUG RESISYANCE GENE IN MYCOBACTERIUM TUBERCULOSIS / 微生物学通报

objective To study the reationship beween drug-resistance gene mutation and drug-resistance level in M. tuberculosis. Methods 108 M. tuberculosis clinical isolated strains from sputum specimens were analyzed by PCR-SSCP and traditional drug susceptibility tests. Results the gene mutation rate of SM, RFP, INH and EMBresistance climical isolated strains was 78.5%, 68.2%, 70.5% and 48.6% respectively, and the mutation rate of SM, RFP, INH and EMB high concentration resistance isolated strains was 86.5% , 89% , 84% and 48.6% respectively, but 28.5 % , 16.6% and 7.1% was the mutation rate of low concentration resistance strains. Conclusion The gene mutation was in relation with drug resistance level of M. tube rculosis. The gene mutation rate was hiher in high concentration resistance isolated strains than in low concentration resistane isolated strains.