| Ulcerative colitis(UC) is an inflammatory bowel disease. Because of slow onset,repeated attack and implied risk of cancerization, and increasing incidence rate year byyear both home and abroad, UC has become one of the hot research areas. In this study,the molecular docking techniques were introduced and combined with the theory andmethods of the discipline of TCM and modern analysis technologies. The experiencedprescription “Wuwei Changyanning” as the research object was investigated, and thestudy focused on the antibacterial and anti-inflammatory chemical constitution ofWuwei Changyanning, its prescription analysis, TNBS/ethanol-induced rats of UCpharmacodynamic model, and multiple indicators of quantitative fingerprint. The mainconclusions were as follows:1)Based on the basic principles and research methods of molecular dockingsimulations,201compositions originated from5herbs and7chemical controlcompositions were obtained through a professional database and some literatures.208compositions were carried out molecular docking with5known antibacterial enzymesand2anti-inflammatory enzymes by using Surflex-dock software (8.0). Relativescore(R, R=the scoring result of medicinal material/the scoring result of chemicalcontrol composition) was used as the evaluation criterion. ⅰ) the scoring results of3compositions, including17-hydroxyhomodapniphyllic acid originated from Niuerfeng,Calyciphylline J originated from Niuerfeng, and Quercetin-3-sulfate originated fromLaliao, were superior to chemical control compositions(R≥1.0). Likewise, the scoringresults of61antibacterial compositions and30anti-inflammatory compositions wereclose to corresponding compared compositions respectively (0.8≤R<1.0). These resultsshowed that the experienced prescription has wide and reliable anti-bacterial andanti-inflammatory chemical constituents, which suggested that the treatment ofCompound Chinese Medicine was multi-targeted and multi-channel.ⅱ) The percentagesof antibacterial and anti-inflammatory compositions originated from5herbs wereobtained via the statistics R≥0.8as follows, respectively. Niuerfeng accounted for37.10%and34.37%, Laliao accounted for25.81%and28.13%, Baishu accounted for4.84%and9.38%, Fuling accounted for12.90%and12.50%, Guanghuoxiangaccounted for19.35%and15.63%. Therefore, the efficiencies of five herbs were rankedas follows:Niuerfeng(Jun drug)>Laliao(Chendrug)>Guanghuoxiang(Shi drug)>Fuling (Zuo drug)>Baishu(Zuo drug). This showed that the efficacies of5herbs were basicallyaccord with “Junchenzuoshi” principle of TCM therapy.2) According to the theory of disassembled prescriptions from TCM, theprescriptions were divided into five groups antimicrobial in vitro, and the animal modelwith anti-inflammatory, strengthening spleen, antidiarrheal, relieving spasm and pain.Prescription analysis showed that:ⅰ) According to the dry herbs, Wuwei Changyanninghad different degrees of inhibition on Escherichia coli (0.1984g/mL), Pseudomonasaeruginosa (0.0248g/mL,), Bacillus proteus (0.0992g/mL), Shigella dysenteriae(0.7936g/mL), Typhoid bacillus(0.3968g/mL) and Staphylococcus aureus (0.0496g/mL).The5herbs had different degrees of antibacterial effect.On the basis of high to low sortby MIC, it was Niuerfeng>Laliao> Guanghuoxiang(except Escherichia coli)>Fulingand Baishu (drug pair). The results between antibacterial activities in vitro andmolecular simulation were basically identical.ⅱ) Compared with the normal controlgroup, Wuwei Changyanning, Niuerfeng and Laliao had very significantlyanti-inflammatory effect (P<0.01), and Guanghuoxiang had significantlyanti-inflammatory effect (p<0.05). On the basis of high to low sort by swelling and theoptical density, it was Niuerfeng>Laliao> Guanghuoxiang>Fuling and Baishu (drugpair). The results between anti-inflammatory effect and molecular simulation werebasically identical. ⅲ)Compared with model group, Wuwei Changyanning andGuanghuoxiang had significantly strengthening spleen (P<0.05) respectively. However,Baishu and Fuling (drug pairs) had very significantly strengthening spleen (p<0.01). ⅳ)Compared with model group, Wuwei Changyanning, Niuerfeng, Laliao, Baishu andFuling(drug pairs) respectively had significantly antidiarrheal effect(p<0.05). ⅴ)Compared with the normal control group,Wuwei Changyanning, Niuerfengand Laliao could very significantly decrease the writhing times of mice (P<0.01) andvery significantly heighten the pain threshold value in60min and120min (P<0.01).Guanghuoxiang could significantly decrease the writhing times of mice (p<0.05) andsignificantly heightene the pain threshold value in60min and120min (p<0.05).However, Baishu and Fuling (drug pairs) only heightened the pain threshold value in120min significantly (p<0.05).3)TNBS/ethanol-induced rats of UC pharmacodynamic model showed that: ⅰ)The general conditions and colon tissue pathomorphology in rats had different degreesof recovery after being treated with high dose, mid dose and low dose from WuweiChangyanning. ⅱ)The contents of TNF-α, IL-8and IL-10in serum of the rats showed that Wuwei Changyanning could decrease proinflammatory cytokines such as TNF-αand IL-8, and increase anti-inflammatory cytokines such as IL-10. The immunologicfunction of rat was restored to normal immune function because Wuwei Changyanningwas inferred that it could keep balance between proinflammatory cytokines andanti-inflammatory cytokines. ⅲ)Wuwei Changyanning Granule could decrease MDA,and increase SOD and GSH-Px, which indicated that reduceing injury andanti-inflammation of the colonic mucosa were achieved.4)The multiple indicator quantitative fingerprint was investigated.ⅰ)The commonpattern of HPLC fingerprint was established including Wuwei Changyanning granuleand5herbs which compose the prescription. Based on the molecular docking resultsand literatures,6compositions including rutin, gallic acid, chlorogenic acid,atractylenolide Ⅰ, pachymic acid and apigenin, which originated from5herbsrespectively, were selected as quantitative compositions. And their contents weredetermined using HPLC from11batches granules and the corresponding5medicinalmaterials.ⅱ) Theprecision, stability and repeatability of fingerprinting wereinvestigated. In addition, common peaks number, the percentage of non-common peaks,and similarity were also studied. The results showed that the identification method offingerprinting was reliable.In summary, the results of molecular docking simulations, disassembledprescription analysis and TNBS/ethanol-induced rats of UC pharmacodynamic modeconsistently indicated that Wuwei Changyanning was an effective compound Chinesemedicine for the treatment of UC. The results of Surflex-Dock molecular docking couldaccurately reflect the true efficacy of the compound Chinese medicine, which is helpfulto effectively reveal the pharmacodynamic mechanism of the chemical composition.Furthermore, it plays a vital role in herbicide efficacy evaluation, pharmacodynamicmechanism and quality control of compound Chinese medicine. The related results andresearch methods can provide a reference for other compound Chinese medicinalpreparation. |
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