ABSTRACT
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.
Subject(s)
Animals , Mice , Anti-Inflammatory Agents/therapeutic use , Colitis, Ulcerative/drug therapy , Colon , Dextran Sulfate/therapeutic use , Drugs, Chinese Herbal , Molecular Docking Simulation , PlasmaABSTRACT
Objective: To investigate the effects of Chinese Dragon’s Blood (CDB) on DSS-induced ulcerative colitis (UC) in mice, and predict the potential main active ingredients, targets and signaling pathways of CDB by network pharmacology. Methods: Mouse UC model was induced by DSS. Balb/c mice were randomly divided into control group, UC group, CDB high, medium, and low dose group, mesalazine control group, 10 mice in each group. Except the control group, each group drank 3.0% DSS solution freely for seven consecutive days. The CDB group and mesalazine group were given corresponding medicines for gastric perfusion during the modeling period. The mice were weighed regularly every day to conduct occult blood test and observe the animals. The fecal traits were evaluated by DAI. At the end of the experiment, the mice were sacrificed and HE stained and scored. The “compound-compound target-disease target” network was constructed by online databases such as TCMSP, String, Cytoscape, etc, and the target targets of core compound targets and compounds were extracted, GO and KEGG enrichment analysis was performed on related targets. Treatment of biological processes and mechanisms of action that UC may regulate were analyzed. Results: Compared with the control group, the infiltration of inflammatory cells in the model group was obvious, and a large number of cup cells disappeared, and the degree of lesions spread to the muscle layer or even the whole layer; The cup cells in the CDB group re-grew, the extent of the lesion was limited to the mucosa, and the inflammation was alleviated. Through the network pharmacology, 112 target targets of CDB were screened, and 14 core compound targets such as ABL1, F2, and JAK2 could be applied to 11 disease targets such as ICAM1, IL-6, PTGS2, and MTOR. The GO analysis contained 415 enrichment pathways, including 389 biological processes, nine molecular functions, and 17 cell components. The KEGG database was used to enrich the relevant pathways, and 84 pathways such as PI3K-Akt were screened for UC. Conclusion: CDB can alleviate colonic mucosal injury induced by DSS in UC mice. It may regulate the expression of ABL1, F2, JAK2 and other target proteins through flavonoids, and then indirectly regulate the expression of IL-6, PTGS2 and other disease proteins. The JAK2/STAT3 and PI3K-Akt-mTOR pathways regulate the levels of inflammatory factors, inhibit the inflammatory response, and ultimately alleviate UC colonic mucosal damage.
ABSTRACT
BACKGROUND: Peripheral nerve injury can lead to extensive changes in central nervous system, and exercise training can promote the recovery of locomotor function following central nervous system injury. OBJECTIVE: To observe the changes of locomotor function and the expression levels of vesicular glutamate transporter VGLUT1 in the spinal cord in a rat model of dorsal root ganglion resection after treadmill exercise and to explore the effect of treadmill training on the locomotor function after peripheral nerve injury. METHODS: Thirty-nine 10-week-old male Wistar rats were randomized into experimental (n=15), control (n=15) and sham operation (n=9) groups. The rats in the experimental and control groups received the dorsal root ganglion resection at L3and L4segments to establish the model of peripheral nerve injury under local anesthesia, while the rats in the sham operation group were only subjected to dorsal root ganglion exposure. The rats in the experimental group underwent 15 m/minute treadmill training at postoperative 7 days, while rats in the other two groups were in free movement. Gait analysis was performed at preoperative 3 days, postoperative 7, 14, 21, and 28 days, respectively, and the behavioral changes of rats were observed. The tissue sections were obtained from L3segment at postoperative 7, 14, and 28 days to detect the expression levels of VGLUT1 in the spinal cord by immunohistochemistry. RESULTS AND CONCLUSION: The peroneal nerve function index in the experimental and control groups was lower than that before surgery and that in the sham operation group at postoperative different time points (all P ＜ 0.05). The index in the experimental and control groups was the lowest on day 7 postoperatively (P ＜ 0.05), then the index gradually increased, but was still lower than the preoperative level (P ＜ 0.05). The index in the experimental group was significantly higher than that in the control group at postoperative 21 and 28 days (P ＜ 0.05). The expression levels of VGLUT1 in the lamina IX in the experimental and control groups were significantly lower than those in the sham operation group at different time points after surgery (P ＜ 0.01). The levels in the experimental group were significantly higher than those in the control group at postoperative 14 and 28 days (P ＜ 0.05). The levels in both groups on a decline after surgery, especially the control group (P ＜ 0.05). These results suggest that treadmill can promote the recovery of locomotor function post peripheral nerve injury.