Introduction
Ulcerative colitis (UC) is a sort of persistent colitis involving colorectal mucosa and submucosa, which is especially brought on by persistent irritation and ulceration. The incidence of UC is growing worldwide,1,2 and its medical manifestations are primarily diarrhea, mucous purulent and bloody stool with belly ache and acute extreme signs. It’s estimated that the whole annual financial burden of UC ranges from $8.1 to $14.9 billion in the US and from €12.5 to €29.1 billion in Europe.3 The period and in depth function of illness are thought-about as essential components related to an elevated danger of colorectal most cancers in sufferers with UC.4–6 Intense efforts have been made to elucidate the pathogeny of UC, however its molecular mechanisms are nonetheless poorly understood. Drug remedy is the primary alternative for UC, and the obtainable medicine embrace mesalazine, corticosteroids, immunosuppressive medicine, and monoclonal antibodies to TNF-α at the moment in Western world.6 Nonetheless, the restrictions of those medicine are additionally apparent, corresponding to poor therapeutic results, excessive value and bothersome negative effects.7
Lately, Conventional Chinese language Drugs (TCM) has been steadily acknowledged as an efficient methodology for treating varied ailments not solely in China and Southeast Asia, but in addition in lots of Western international locations.8,9 TCM additionally performs an essential position in drug growth, offering plentiful novel compound lead sources. Huangtu Decoction (HTD) was first recorded within the Treatise on “JingGuiYaoLue (Synopsis of Golden Chamber)” written round 205 A.D by Zhang Zhongjing, a well-known medical scientist within the Japanese Han Dynasty. Its prescription consists of seven medicinal herbs, together with Terra Flava Usta, Rehmanniae Radix, Scutellariae Radix, Aconiti Lateralis Radix Praeparata, Asini Corii Colla, Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma. HTD has been primarily used within the remedy of bleeding signs, corresponding to blood below stool, hematemesis, bleeding, metrorrhagia and boring blood colour, and extremely revered by medical doctors for hundreds of years in China.10,11 In fashionable instances, HTD is commonly used to deal with gastrointestinal bleeding, persistent UC, gastrointestinal tumor and so forth.12,13 Nevertheless, the energetic compounds and mechanism of HTD for the remedy of UC are nonetheless not clear.
Over the past a long time, bioinformatics evaluation was broadly used to display genetic alterations on the genome degree,14,15 in order to determine the differentially expressed genes (DEGs). Apart from, community pharmacology has been efficiently launched to disclose the mechanisms of TCM, which coincides with the connotation of multi-component, multi-targets, and multi-pathways holistic technique.16,17
Within the current research, community pharmacology, bioinformatics evaluation and in vivo experiments have been mixed to uncover the potential targets and mechanisms of HTD for treating UC, and the detailed flowchart of the research design is proven in Figure 1.
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Determine 1 Flowchart of the evaluation technique within the research. |
Supplies and Strategies
Reagents and Supplies
Terra Flava Usta (Batch No: 2006002) was bought from AnguoJuyaotang Pharmaceutical Co., Ltd. Rehmanniae Radix (Batch No: Y2105018), Scutellariae Radix (Batch No: Y2007053), Aconiti Lateralis Radix Praeparata (Batch No: Y062011002), Asini Corii Colla (Batch No: Y052105035), Atractylodis Macrocephalae Rhizoma (Batch No: Y2006043) and Glycyrrhizae Radix et Rhizoma (Batch No: Y2008055) have been bought from Sichuan Neautus Conventional Chinese language Drugs Co., Ltd. HPLC grade methanol was supplied by Fisher Scientific (USA). Different chemical compounds have been of analytical grade.
Animals and Diets
The C57BL/6 male mice have been supplied by the SPF (Beijing) Biotechnology Co., Ltd. (DCXK (Beijing) 2019–0010). The mice (6–8 weeks previous, n=24) have been randomly divided into 3 teams, and acclimated in impartial air flow cages (IVC) for six days with free entry to straightforward mouse chow and faucet water within the SPF facility below managed temperature (22 ± 2°C), humidity (55% ± 10%), and 12 light-dark cycle. Animal experiments have been carried out in response to the Advisable Tips for the Care and Use of Laboratory Animals issued by the Chinese language Council on Animal Analysis and authorised by the Ethics Committee of Chengdu College of TCM.
Preparation of the HTD Extracts
The HTD extracts have been ready in response to the recorded methodology in “JingGuiYaoLue”. Briefly, the seven herbs Terra Flava Usta (40 g), Rehmanniae Radix (15 g), Scutellariae Radix (15 g), Aconiti Lateralis Radix Praeparata (15 g), Asini Corii Colla (15 g), Atractylodis Macrocephalae Rhizoma (15 g) and Glycyrrhizae Radix et Rhizoma (6 g) have been blended with purified water, and Aconiti Lateralis Radix Praeparata have been soaked for 12 hours, whereas different herbs have been soaked for half an hour, then they have been boiled for an hour. The filtrate have been collected and centrifuged at 3000 rpm for five min. The supernatant was harvested and steamed dry, and the residue was dissolved to five mL with methanol. The pattern was filtered by way of a 0.22 μm microporous membrane for subsequent testing.
Identification of Lively Compounds in HTD
The energetic compounds of HTD extracts have been qualitatively recognized by Q Exactive Orbitrap LC-MS/MS (Thermo Fisher Scientific, Waltham, MA, USA). The XBridgeBEHC18 (2.1×100 mm, i.d.; 2.5 μm, Waters, MA, USA) was utilized for pattern separation at 35°C. Methanol (A) and 0.1% formic acid-water (B) (95:5) have been used as cellular section. Detection wavelength was set at 280 nm. The circulate price and injection quantity have been set as 0.2 mL/min and 1.0 μL, respectively. Nitrogen was utilized as auxiliary fuel and sheath fuel at a circulate price of 12 L/min. The mass willpower was carried out primarily based on optimistic and damaging scanning mode with the m/z of 100–1000.
Acquisition of Corresponding Targets of Lively Compounds
All of the chemical constructions of the recognized energetic compounds have been imported into SwissTargetPrediction (http://www.swisstargetprediction.ch/) to foretell the targets.18 Targets of those compounds have been collected by way of high-throughput screening and reverse docking.
Acquisition of UC Targets
Gene Expression Omnibus (GEO) (https://www.ncbi.nlm.nih.gov/geo), a public practical genomics information repository, was used to determine UC targets library.19 Key phrases corresponding to “ulcerative colitis” or “ulcer colonitis” have been used to seek for the identified targets associated to the pathogenesis of UC from GEO database, in order to seek out out the distinction in gene expression datasets between intestinal tissue samples from sufferers with UC and regular intestinal tissue samples. Lastly, 3 datasets of UC have been downloaded from the GEO database: GSE75214 on the (HuGene-1_0-st) Affymetrix Human Gene 1.0 ST Array (GPL6244 platform), together with colon tissue samples from 97 UC sufferers and 11 wholesome folks; GSE87466 on the (HT_HG-U133_Plus_PM) Affymetrix HT HG-U133+ PM Array Plate (GPL13158 platform), together with colon tissue samples from 87 UC sufferers and 21 wholesome folks; GSE59071 on the (HuGene-1_0-st) Affymetrix Human Gene 1.0 ST Array (GPL6244 platform), together with colon tissue samples from 97 UC sufferers and 11 wholesome folks.
The datasets downloaded from the GEO database have been preprocessed by background correction, quantile normalization and base logarithm conversions. Lastly, the distinction in gene expression between regular human and UC sufferers was clarified. On this research, log FC (fold change) ≥1 and adjusted p-value <0.01 thought-about that the distinction was statistically vital, and the corresponding genes have been recognized as DEGs, listed within the UC goal library.
GO and Pathway Enrichment Evaluation
The GO and pathway enrichment evaluation was carried out by way of Metascape (https://metascape.org).20 GO functionally annotates key genes into 3 important phrases, ie mobile elements (CCs), molecular features (MFs), and organic processes (BPs). Apart from, CytoscapeClueGO plugin was used to additional analyze the enrichment of BPs. Pathway enrichment evaluation unveiled the potential BPs with key targets. As well as, the bubble chart of GO and pathway enrichment evaluation have been carried out on the bioinformatics platform (http://www.bioinformatics.com.cn/).
Development of Compound-Goal-Pathway Community
To characterize the therapeutic mechanisms of HTD for UC, the compound-target-pathway community was constructed utilizing Cytoscape model 3.7.2.21 Within the community, the nodes with totally different colours and shapes represented the drug, compounds, illness, goal genes, or disease-related pathways, respectively, and an “edge” was an affiliation between the nodes.
Experimental Validation
Dextran Sodium Sulfate (DSS)-Induced Acute UC Mannequin
Following the earlier strategies of Mariano et al and Meurer et al22,23 after adaptive feeding for six days, the mice in management group (n=8) have been fed with water, whereas these in mannequin group (n=8) and DSS + HTD group (n=8) freely drank 3% DSS (36,000–50,000 Da) resolution (ready by dissolving 3 g of DSS in 100 mL of deionized water). The consuming quantity in every group was recorded, and DSS resolution was up to date every single day. Moreover, the mice in DSS + HTD group have been administrated with 2.5 g/kg HTD extracts as soon as a day in response to their weight. The consuming quantity, physique weight, stool properties and blood in stool of mice in every group have been recorded every single day, and the colitis scores got by weight reduction and stool circumstances proven in Table 1. After investigations for one week, the mice within the 3 teams have been euthanized, and the serum and colon tissues have been collected for additional evaluation.
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Desk 1 Scoring Requirements of Ulcer Colitis |
Enzyme-Linked Immunosorbent Assay (ELISA)
The degrees of inflammatory components have been detected by ELISA kits. The mouse IL-1β ELISA equipment (Multi Sciences (Lianke) Biotech, CO., LTD., Hangzhou, China), and mouse IL-6 ELISA equipment (Multi Sciences (Lianke) Biotech, CO., LTD., Hangzhou, China) have been utilized in accordance with the instructions of the manufacture.
Histopathology
The colon samples have been minimize into small items, mounted in 4% paraformaldehyde, embedded in paraffin, and stained with hematoxylin and eosin (H&E). Then histopathological adjustments have been examined below a microscope.
Western Blotting (WB)
Proteins have been extracted from the colon tissues utilizing RIPA Lysis Buffer (Wuhan Servicebio Expertise Co., Ltd, China) and the protein focus was decided by bicinchoninic acid (BAC) methodology on the enzyme labelling instrument (Thermo Fisher Scientific, USA). The SDS-PAGE was used to separate the protein, and subsequently transferred to polyvinylidene fluoride (PVDF) membranes. The membranes have been first blocked for 1h with 5% BSA at room temperature, after which probed with major antibodies of MMP1 (Affinity, dilution of 1:1000), MMP3 (Abcam, dilution of 1:5000), MMP7 (Abcam, dilution of 1:1000), MMP9 (Abcam, dilution of 1:1000), MMP12 (Proteintech, dilution of 1:1000) and β-Actin (Affinity, dilution of 1:5000) at 4°C for in a single day after which incubated with HPR-conjugated antibody at 37°C for one hour. Lastly, the protein bands have been recorded by ELC (Thermo Fish Scientific, USA) and quantified by Picture J software program to judge the protein relative expressions examine with β-actin.
Quantitative Actual-Time- PCR (qRT-PCR)
The mRNA expression of UC was detected by qRT-PCR. The colon tissues in every group of mice have been pulverized in liquid nitrogen earlier than RNA extraction. The full RNA was remoted and reversely transcribed into cDNA utilizing the RT-qPCR SimpleTM (One Step)-SYBR Inexperienced I equipment (Foregene; Chengdu, China), adopted by synthesis by way of reverse transcription utilizing the Direct RT Combine within the equipment, and amplification utilizing Direct qPCR Combine-SYBR of equipment in response to the producer’s directions. Then qPCR was carried out on a Bio-Rad real-time PCR system (CFXConnect, USA) for 40 cycles. The expression ranges of goal genes have been normalized to the management housekeeping gene GAPDH. The gene expression was analyzed by the two−ΔΔCt methodology. Primer sequences are proven in Table 2.
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Desk 2 Primer Sequences for qRT-PCR |
Immunohistochemistry and Staining Depth
Immunolabeling for matrix metalloproteinase-1 (MMP1), MMP3, MMP7, MMP9 and MMP12 was carried out on mounted and paraffin-embedded sections. Briefly, following deparaffinization and rehydration, slides have been subjected to antigen retrieval in citrate buffer (pH 6.0) for 20 min in a steamer, rinsed in PBS (0.01 mol/L phosphate buffer containing 0.15 mol/L NaCl, pH7.4), and incubated for 10 min in 3% H2O2 to dam endogenous peroxidases. After that, the sections have been incubated with anti-MMP1 (1:200; Bioss, China), anti-MMP3 (1:200; Bioss, China), anti-MMP7 (1:200; Bioss, China), anti-MMP9 (1:200; Bioss, China), and anti-MMP12 (1:200; Bioss, China) major antibodies at 4°C in a single day in a moist field, after which with HRP-labeled anti-rabbit secondary antibodies for 30 min, adopted by counterstaining with hematoxylin, rehydration and mounting. Staining depth was quantified by Picture-Professional Plus 6.0 (Media Cybernetics) and expressed as imply utilizing the system “imply density = built-in optical density/space of curiosity”.
Statistical Evaluation
Every experiment was repeated 3 instances, and the outcomes have been expressed as imply ± commonplace deviation (SD). Vital variations between means have been in contrast utilizing the Bonferroni and Dunnett’s T3 take a look at within the case of homogeneity of variance with the p-value <0.05.
Outcomes
Compounds of HTD Extract and Corresponding Targets
The primary compounds of HTD extracts have been analyzed qualitatively primarily based on Q Exactive Orbitrap LC-MS/MS. The full ion chromatogram (TIC) is proven below positive-ion polarity mode (A) and negative-ion polarity mode (B) in Figure 2.
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Determine 2 Chromatograph of the extracts of HTD by Q Exactive Orbitrap LC-MS/MS on positive-ion polarity mode (A) and negative-ion polarity mode (B). |
Greater than 60 main compounds have been eluted by the Q Exactive Orbitrap LC-MS/MS system from the HTD extracts inside 30 min. Amongst then, a complete of 47 compounds have been lastly recognized by evaluating with the identified chemical constituents within the reported literature information, and deduced in response to their mass spectrometry and fragment ion traits, as proven in Table 3 and Figure 3.
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Desk 3 Compositions and Their Product Ions in Huangtu Decoction |
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Determine 3 Compounds detected within the extracts of HTD by Q Exactive Orbitrap LC-MS/MS. |
A complete of 511 corresponding targets of compounds have been acquired from Swiss Goal Prediction.
Prediction of Targets of HTD for UC
After information preprocessing, a complete of 470 DEGs have been extracted from 3 datasets (GSE87466, GSE59071, and GSE75214) downloaded from the GEO database, as proven within the volcano map and scorching map (Figure 4A and B). Lastly, 154 upregulated and 316 downregulated genes have been recognized in UC tissues in contrast with regular colon tissues.
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Determine 4 The overlapped targets screening technique. (A) The volcano plot of the differentially expressed genes (DEGs) in datasets of GSE87466, GSE59071, and GSE75214.The pink nodes characterize up-regulated DEGs, whereas inexperienced nodes characterize down-regulated DEGs; the horizontal dashed strains present the P worth is lower than 0.05 and the vertical dashed strains point out the worth of /log2 fold change (FC)/ is greater than 1. (B) Heatmap of DEGs. The vertical axis represents the genes. The horizontal axis reveals the change of expression ranges of DEGs in datasets of GSE87466, GSE59071, and GSE75214 by -log10Pvalue. (C) Overlap genes of DEGs and predicted compound targets. |
Lastly, a complete of 29 overlapping genes (OGs) of compound targets and UC-related targets, particularly HSD17B2, KDR, PLAU, ABCG2, MMP7, NOS2, MMP3, FADS1, CXCR1, AKR1B10, CDC25B, SELL, CD38, MAOA, EDNRA, LRRK2, CA2, ABCB1, STS, PDGFRB, SELP, PTGS2, PFKFB3, PIM2, MMP1, IGFBP5, EPHX2, MMP12 and MMP9, have been considered potential targets of HTD for the remedy of UC (Figure 4C).
GO and Enrichment Evaluation
As proven in Figure 5A and Table 4, the highest phrases (P<0.01) within the BPs, CCs, and MFs have been chosen. The enrichment of BPs concerned the collagen catabolic course of, collagen metabolic course of, extracellular matrix disassembly, regulation of neuroinflammatory response, extracellular matrix group, extracellular construction group, exterior encapsulating construction group, response to oxidative stress, reactive oxygen species metabolic course of, and response to inorganic substance. Apart from, additional evaluation of the community was carried out by Cytoscape ClueGO plugin, and it was discovered that the collagen catabolic course of, optimistic regulation of clean muscle cell proliferation, long-chain fatty acid biosynthetic course of, and optimistic regulation of phospholipase exercise have been primarily concerned in BPs (Figure 5B). The enrichment of CCs primarily concerned secretory granule membrane, and different cell elements. The enrichment of MFs concerned the metalloendopeptidase exercise and metallopeptidase exercise.
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Desk 4 GO Practical Enrichment Evaluation Outcomes of Overlapping Genes (OGs) |
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Determine 5 Bubble diagram of GO practical enrichment evaluation of overlapped genes (A) The bubble chart. (B) The visualization evaluation of BPs. |
The sign transduction pathways are proven in Table 5 and Figure 6 (P<0.01). The results of pathway enrichment evaluation of the HTD for UC revealed that the OGs have been considerably related to MMPs, imatinib, persistent myeloid leukemia, and photodynamic therapy-induced NF-κB survival signaling, in WikiPathways, with IL-4 and IL-13 signaling and collagen degradation in Reactome Gene Units, and likewise with MicroRNAs in most cancers and IL-17 signaling pathway in KEGG.
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Desk 5 Pathway Enrichment Evaluation Outcomes of Overlapping Genes (OGs) |
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Determine 6 Enriched pathway evaluation of potential targets of HTD compounds towards UC. |
Compound-Goal- Pathway Community
As proven in Figure 7, the compound-target-pathway community included 46 nodes (11 compounds, 18 targets, and 17 pathways) and 158 edges. Based on the evaluation of the community, every energetic compound acted on not less than 2 goal genes, every gene was regulated by not less than 2 energetic compounds, and not less than 18 genes have been doubtlessly concerned in every pathway associated to UC, indicating the motion traits of a number of energetic compounds and targets of HTD within the remedy of UC.
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Determine 7 Compound-Goal- Pathway Community, the pink triangle node are the sign transduction pathways; inexperienced hexagon nodes are OGs, blue rectangle nodes are the compounds of HTD extract, strains characterize the interactions between them. |
Experimental Validation
Impact of HTD on Histopathological Modifications
As proven in Figure 8A and B, the mice in mannequin group and DSS+HTD group manifested various levels of free stool and weight reduction in contrast with these in management group. The pathological circumstances of colon tissues of the mice handled with DSS+HTD have been improved in contrast with these in mannequin group. The histopathological adjustments in every group are proven in Figure 8C. The outcomes of H&E staining confirmed that DSS induced huge inflammatory infiltration, mucosal edema, and hyperplasia in colonic tissues of mannequin group, however much less necrosis was noticed in DSS+HTD group.
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Determine 8 HTD ameliorates DSS-induced colitis. (A) Curve of every day Colitis scores. (B) Curve of every day physique weights. (C) Colonic mucosa of regular group mice. (D) Colon of a DSS-treated mouse, extreme inflammatory infiltration, edema, lack of crypts and ulcerations are seen. (E) HTD decreased DSS-induced colonic irritation. (F and G) Impact of HTD on serum degree of IL-1βand IL-6. Information are expressed as imply±SD (n = 8). *P < 0.05, **P < 0.01 and ***P < 0.001 versus mannequin group. |
Impact of HTD on Serum Ranges of IL-1β and IL-6
As proven in Figure 8F and G, the serum ranges of IL-1β and IL-6 have been considerably elevated in mannequin group in contrast with these in management group (P<0.01). In comparison with the mannequin group, serum IL-1β and IL-6 ranges decreased significantly in DSS+HTD group (P<0.01).
Impact of HTD on MMP Expressions
The expression ranges of MMP1, MMP3, MMP7, MMP9 and MMP12 in colon tissues have been investigated by way of immunohistochemical staining (Figure 9) and WB (Figure 10A and B). In distinction with mannequin group, the expressions of MMP1, MMP3 and MMP9 in colon tissues in DSS+HTD group have been considerably decrease, and the expressions of MMP7 and MMP12 declined with no vital distinction. As well as, qRT-PCR was utilized to discover the position of HTD in regulating the expression of MMPs in colon tissues. As proven in Figure 10C, DSS considerably upregulated the expressions of MMP1, MMP3, MMP7, MMP9 and MMP12 in colon tissues, however after HTD remedy, the upregulation of MMP1, MMP3 and MMP9 was reversed, whereas MMP7 and MMP12 had no vital adjustments.
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Determine 9 Consultant immunohistochemical staining with MMP1, MMP3, MMP7, MMP9 and MMP12 in colon tissues in mice handled with DSS. (A) Immunohistochemical staining for MMP1, MMP3, MMP7, MMP9 and MMP12 in regular, mannequin and DSS+HTD group. (B) Statistical evaluation of relative immunohistochemical staining depth for MMP1, MMP3, MMP7, MMP9 and MMP12 in regular, mannequin and DSS+HTD group.*P < 0.05, **P < 0.01 and ***P < 0.001 versus mannequin group. |
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Determine 10 Results of HTD on the expression of MMPs. (A) Western blot of MMP1, MMP3, MMP7, MMP9 and MMP12 in colon tissues. (B) Quantification of MMP1, MMP3, MMP7, MMP9 and MMP12. (C) Decreased protein expressions of MMP1, MMP3 MMP7, MMP9 and MMP12 within the colon obtained from mice handled with DSS. *P < 0.05, **P < 0.01 and ***P < 0.001 versus mannequin group. |
Dialogue
UC is a persistent illness with an growing incidence worldwide over current decade, particularly in creating nations.2 HTD is a really efficient prescription for the medical remedy of hematochezia in TCM, and has been reported as non-surgical remedy of UC with minimal negative effects. Nevertheless, there’s a lack of contemporary animal experimental proof and no literature report on its energetic compounds and mechanism. On this research, the UC mannequin was established to judge the therapeutic results of HTD on UC, and the potential mechanism of this TCM system for treating UC was additionally explored primarily based on community pharmacology.
HTD incorporates 7 medicinal herbs, together with Rehmanniae Radix, Scutellariae Radix, Aconiti Lateralis Radix Praeparata, Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma, Terra Flava Usta and Asini Corii Colla. Within the current research, HPLC-MS/MS was used to research the chemical compositions of HTD, and a complete of 47 compounds have been recognized from it, largely flavones and alkaloids. Moreover, community pharmacology was used to discover its potential mechanism. The compound-target-pathway community confirmed that wogonin with the very best diploma was considered the simplest compound, adopted by LicoricesaponinG2, Liquiritigenin, Retrochalcone, Genistein, Atractylenolide III, Glycyrrhizic acid, Pinobanksin, Pinocembrin, Mussaenosidic acid and Hesperetin. These compounds recognized on this system could present fundaments for establishing Q-markers for additional dependable high quality management of HTD. As well as, the potential molecular mechanism and signaling pathway for HTD have been additionally predicted primarily based on GO and KEGG enrichment evaluation. The outcomes revealed that the enrichment of BPs of HTD concerned collagen catabolic course of, collagen metabolic course of, and extracellular matrix disassembly. The pathway enrichment evaluation outcomes confirmed that the pathways coated MMPs, MicroRNAs in most cancers, IL-17 signaling pathway and so forth. Subsequently, the prediction outcomes have been additional verified by in vivo animal experiments, and an identical outcome to community pharmacology was obtained. HTD was efficient within the remedy of UC by way of lowering the protein and mRNA expressions of MMP1, MMP3 and MMP9. MMPs are a household of proteolytic enzymes characterised by the flexibility to degrade extracellular matrix (ECM) proteins corresponding to collagen, proteoglycan, fibronectin, elastin, and laminin.41,42 Continual irritation and aberrant tissue transforming with extreme accumulation or degradation of ECM elements are markers in UC. Therefore, MMPs are regarded as predominant proteases concerned within the pathogenesis of UC.41–43 MMPs have been acknowledged as noninvasive diagnostic instruments for UC in lots of research.41,42 Particular artificial MMP inhibitors and a few pure merchandise or therapeutic interventions have been described to ameliorate intestinal irritation.43
Conclusion
In conclusion, HTD can alleviate the colonic irritation by way of inhibiting MMPs together with MMP1, MMP3 and MMP9. The current research will profit the additional growth of this basic TCM system.
Acknowledgments
The authors want to thank the editor and the nameless reviewers whose constructive feedback are very useful in strengthening the presentation of this paper. The analysis was supported by Chengdu Excessive-level Key Medical Specialty Development Venture.
Disclosure
The authors report no conflicts of curiosity on this work.
References
1. Du L, Ha C. Epidemiology and pathogenesis of ulcerative colitis. Gastroenterol Clin North Am. 2020;49(4):643–654. doi:10.1016/j.gtc.2020.07.005
2. da Silva BC, Lyra AC, Rocha R, et al. Epidemiology, demographic traits and prognostic predictors of ulcerative colitis. World J Gastroenterol. 2014;20(28):9458–9467. doi:10.3748/wjg.v20.i28.9458
3. Cohen RD, Yu AP, Wu EQ, et al. Systematic evaluation: the prices of ulcerative colitis in Western international locations. Aliment Pharmacol Ther. 2010;31(7):693–707. doi:10.1111/j.1365-2036.2010.04234.x
4. Chen R, Lai LA, Brentnall TA, et al. Biomarkers for colitis-associated colorectal most cancers. World J Gastroenterol. 2016;22(35):7882–7891. doi:10.3748/wjg.v22.i35.7882
5. Nehme F, Schneider A, Hamid F. Appendiceal adenocarcinoma related to ulcerative colitis. ACG Case Rep J. 2019;6(11):e00255. doi:10.14309/crj.0000000000000255
6. Ordás I, Eckmann L, Talamini M, et al. Ulcerative colitis. Lancet. 2012;380(9853):1606–1619. doi:10.1016/s0140-6736(12)60150-0
7. Wehkamp J, Stange EF. Latest advances and rising therapies within the non-surgical administration of ulcerative colitis. F1000 Res. 2018;7:1207. doi:10.12688/f1000research.15159.1
8. Zhang Y, Lengthy Y, Yu S, et al. Pure unstable oils derived from natural medicines: a promising remedy manner for treating depressive dysfunction. Pharmacol Res. 2021;164:105376. doi:10.1016/j.phrs.2020.105376
9. Xu Q, Guo Q, Wang C, et al. Community differentiation: a computational methodology of pathogenesis prognosis in conventional Chinese language drugs primarily based on techniques science. Artif Intell Med. 2021;118:102134. doi:10.1016/j.artmed.2021.102134
10. Shao MJ, Yan YX, Qi Q, Tang W, Zuo JP. Utility of energetic elements from conventional Chinese language drugs in remedy of inflammatory bowel illness. Zhongguo Zhong Yao Za Zhi. 2019. 44(3):415–421. doi:10.19540/j.cnki.cjcmm.20180907.001. Chinese language. PMID: 30989902.
11. Liu F, Liu Y, Tian C. Impact of Rhizoma Atractylodis extract in defending gastric mucosa and modulating gastrointestinal immune perform in a rat mannequin of spleen deficiency. Nan Fang Yi Ke Da Xue Xue Bao. 2015;35(3):343–7, 354. Chinese language. PMID: 25818777.
12. Tang XD, Lu B, Li ZH, et al. Therapeutic impact of Chang’an I recipe (I) on irritable bowel syndrome with diarrhea: a multicenter randomized Double-Blind placebo-controlled medical trial. Chin J Integr Med. 2018;24(9):645–652. doi:10.1007/s11655-016-2596-9
13. Zhang Q, Duan H, Li R, et al. Inducing apoptosis and suppressing inflammatory reactions in synovial fibroblasts are two essential methods for Guizhi-Shaoyao-Zhimu decoction towards rheumatoid arthritis. J Inflamm Res. 2021;14:217–236. doi:10.2147/JIR.S287242
14. Manfredi M, Brandi J, Di Carlo C, et al. Mining most cancers biology by way of bioinformatic evaluation of proteomic information. Knowledgeable Rev Proteomics. 2019;16(9):733–747. doi:10.1080/14789450.2019.1654862
15. Tang S, Jing H, Huang Z, et al. Identification of key candidate genes in neuropathic ache by built-in bioinformatic evaluation. J Cell Biochem. 2020;121(2):1635–1648. doi:10.1002/jcb.29398
16. Li ZT, Zhang FX, Fan CL, et al. Discovery of potential Q-marker of conventional Chinese language drugs primarily based on plant metabolomics and community pharmacology: periplocae cortex for example. Phytomedicine. 2021;85:153535. doi:10.1016/j.phymed.2021.153535
17. Pan L, Li Z, Wang Y, et al. Community pharmacology and metabolomics research on the intervention of conventional Chinese language drugs Huanglian decoction in rats with sort 2 diabetes mellitus. J Ethnopharmacol. 2020;258:112842. doi:10.1016/j.jep.2020.112842
18. Daina A, Michielin O, Zoete V. SwissTargetPrediction: up to date information and new options for environment friendly prediction of protein targets of small molecules. Nucleic Acids Res. 2019;47(W1):W357–W364. doi:10.1093/nar/gkz382
19. Clough E, Barrett T. The gene expression omnibus database. Strategies Mol Biol. 2016;1418:93–110. doi:10.1007/978-1-4939-3578-9_5
20. Zhou Y, Zhou B, Pache L, et al. Metascape supplies a biologist-oriented useful resource for the evaluation of systems-level datasets. Nat Commun. 2019;10(1):1523. doi:10.1038/s41467-019-09234-6
21. Doncheva NT, Morris JH, Gorodkin J, et al. Cytoscape StringApp: community evaluation and visualization of proteomics information. J Proteome Res. 2019;18(2):623–632. doi:10.1021/acs.jproteome.8b00702
22. Mariano LNB, Arruda C, Somensi LB, et al. Brazilian inexperienced propolis hydroalcoholic extract reduces colon damages brought on by dextran sulfate sodium-induced colitis in mice. Inflammopharmacology. 2018;26(5):1283–1292. doi:10.1007/s10787-018-0467-z
23. Meurer MC, Mees M, Mariano LNB, et al. Hydroalcoholic extract of Tagetes erecta L. flowers, wealthy within the carotenoid lutein, attenuates inflammatory cytokine secretion and improves the oxidative stress in an animal mannequin of ulcerative colitis. Nutr Res. 2019;66:95–106. doi:10.1016/j.nutres.2019.03.005
24. Farooq SM, Hou Y, Li H, et al. Disruption of GPR35 exacerbates dextran sulfate sodium-induced colitis in mice. Dig Dis Sci. 2018;63(11):2910–2922. doi:10.1007/s10620-018-5216-z
25. Tune W, Qiao X, Chen Ok, et al. Biosynthesis-based quantitative evaluation of 151 secondary metabolites of licorice to distinguish medicinal Glycyrrhiza species and their hybrids. Anal Chem. 2017;89(5):3146–3153. doi:10.1021/acs.analchem.6b04919
26. Tanaka Ok, Hayashi Ok, Fahad A, Arita M. Multi-stage mass spectrometric evaluation of saponins in Glycyrrhiza radix. Nat Prod Commun. 2011;6(1):7–10. PMID: 21366035.
27. Chang GH, Bo YY, Cui J, et al. Major chemical constituents in aerial elements of Glycyrrhiza uralensis by UPLC-Q-Exactive Orbitrap-MS. Zhongguo Zhong Yao Za Zhi. 2021;46(6):1449–1459. doi:10.19540/j.cnki.cjcmm.20201225.301
28. Fong SY, Wong YC, Zuo Z. Improvement of a SPE-LC/MS/MS methodology for simultaneous quantification of baicalein, wogonin, oroxylin A and their glucuronides baicalin, wogonoside and oroxyloside in rats and its software to mind uptake and plasma pharmacokinetic research. J Pharm Biomed Anal. 2014;97:9–23. doi:10.1016/j.jpba.2014.03.033
29. Chai CC, Cao Y, Mao M, et al. Comparability of chemical compositions earlier than and after wine-frying of Scutellaria baicalensis primarily based on HPLC attribute chromatogram, UPLC-Q-TOF/ MS qualitative and multi-component quantitative evaluation. Chin Tradit Herb Medicine. 2020;51(9):2436–2447. doi:10.7501/j.issn.0253-2670.2020.09.019
30. Wei Y, Wang SY, Wu SY, et al. Qualitative characterization of flavonoids in Scutellariae Radix by utilizing PREC-IDA-EPI. Zhongguo Zhong Yao Za Zhi. 2018;43(2):345–352. doi:10.19540/j.cnki.cjcmm.20171027.004
31. Cui YY, Zhou YF, Ma YQ, et al. Variations evaluation of chemical composition of uncooked and fried Glycyrrhiza uralensis primarily based on UPLC-QTOF-MS. Chin Pharm. 2020;31(9):1049–1053. doi:10.6039/j.issn.1001-0408.2020.09.06
32. Solar X, Wen HM, Cui XB, et al. Qualitative analysis of Atractylodis Macrocephalae rhizoma from totally different habitats by HPLC-PDA fingerprint mixed with UFLC-Q-TOF/MS qualitative identification. Chin Tradit Herb Medicine. 2016;47(19):3494–3501. doi:10.7501/j.issn.0253-2670.2016.19.023
33. Cao G, Li Q, Cai H, et al. Investigation of the chemical adjustments from crude and processed Paeoniae Radix Alba-Atractylodis Macrocephalae rhizoma natural pair extracts by utilizing Q exactive high-performance benchtop quadrupole-orbitrap LC-MS/MS. Evid Based mostly Complement Alternat Med. 2014;2014:170959. doi:10.1155/2014/170959
34. Zhang BY, Jiang ZZ, Wang YF, et al. Evaluation of chemical constituents in contemporary, dried and ready Rehmanniae Radix by UPLC/ESI-Q-TOF MS. Chin Tradit Patent Med. 2016;38(5):1104–1108. doi:10.3969/j.issn.1001-1528.2016.05.029
35. Qiao X, Li R, Tune W, et al. A focused technique to research untargeted mass spectral information: speedy chemical profiling of Scutellaria baicalensis utilizing ultra-high efficiency liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry and key ion filtering. J Chromatogr A. 2016;1441:83–95. doi:10.1016/j.chroma.2016.02.079
36. Wang L, Tan N, Wang H, et al. A scientific evaluation of pure alpha-glucosidase inhibitors from flavonoids of Radix scutellariae utilizing ultrafiltration UPLC-TripleTOF-MS/MS and community pharmacology. BMC Complement Med Ther. 2020;20(1):72. doi:10.1186/s12906-020-2871-3
37. Zhang DK, Han X, Rui YL, et al. Evaluation on attribute constituents of crude Aconitum carmichaelii in several areas primarily based on UPLC-Q-TOF-MS. Zhongguo Zhong Yao Za Zhi. 2016;41(3):463–469. doi:10.4268/cjcmm20160318
38. Ye X, Wu J, Zhang D, et al. How Aconiti Radix cocta can deal with gouty arthritis primarily based on systematic pharmacology and UPLC-QTOF-MS/MS. Entrance Pharmacol. 2021;12:618844. doi:10.3389/fphar.2021.618844
39. Zhou SS, Ma ZC, Liang QD, et al. UPLC/Q-TOF-MS-based chemical profiling method to judge the chemical structure of Radix Aconiti lateralis preparata within the means of decoction. Zhong Xi Yi Jie He Xue Bao. 2012;10(8):894–900. doi:10.3736/jcim20120810
40. Zhu H, Wang C, Qi Y, et al. Fingerprint evaluation of Radix Aconiti utilizing ultra-performance liquid chromatography-electrospray ionization/ tandem mass spectrometry (UPLC-ESI/MS n) mixed with stoichiometry. Talanta. 2013;103:56–65. doi:10.1016/j.talanta.2012.10.006
41. O’Shea NR, Smith AM. Matrix metalloproteases position in bowel irritation and inflammatory bowel illness: an updated evaluation. Inflamm Bowel Dis. 2014;20(12):2379–2393. doi:10.1097/MIB.0000000000000163
42. Maronek M, Marafini I, Gardlik R, et al. Metalloproteinases in inflammatory bowel ailments. J Inflamm Res. 2021;14:1029–1041. doi:10.2147/JIR.S288280
43. de Bruyn M, Vandooren J, Ugarte-Berzal E, et al. The molecular biology of matrix metalloproteinases and tissue inhibitors of metalloproteinases in inflammatory bowel ailments. Crit Rev Biochem Mol Biol. 2016;51(5):295–358. doi:10.1080/10409238.2016.1199535
