Effect of Huangqin Tang on Colonic Gene Expression in Rats with Ulcerative Colitis

Wang, Dunfang; Shi, KaiFeng; Wang, Yanli; Zou, Dixin; Guo, Shanshan; Li, Tao; Xu, Hangyu; Ma, Xuran; Liu, JiaXing; Song, HongXin; Yang, Weipeng; Li, Yu

doi:https://doi.org/10.1155/2020/4238757

International Journal of Genomics

On this page

Abstract Introduction Materials and Methods Results Discussion Data Availability Conflicts of Interest Authors’ Contributions Acknowledgments References Copyright Related Articles

Research Article | Open Access

Volume 2020 | Article ID 4238757 | https://doi.org/10.1155/2020/4238757

Effect of Huangqin Tang on Colonic Gene Expression in Rats with Ulcerative Colitis

Dunfang Wang,¹KaiFeng Shi,²Yanli Wang,¹Dixin Zou,¹Shanshan Guo,¹Tao Li,³Hangyu Xu,¹Xuran Ma,¹JiaXing Liu,¹HongXin Song,¹Weipeng Yang,¹and Yu Li⁴

Academic Editor: Ernesto Picardi

Received17 Oct 2019

Revised24 Dec 2019

Accepted13 Jan 2020

Published27 Mar 2020

Abstract

In this study, we explored the pharmacological mechanisms of Huangqin Tang (HQT; a traditional Chinese medicine formula) in ulcerative colitis (UC) and provided evidence for potential roles HQT plays by gene expression profiling. The UC rat model was made via a compound method (trinitrobenzene sulfonic acid plus ethanol). After a ten-day treatment, microarray analysis was performed from the colon segment of the rats. Biological functions and specific signaling pathways were enriched based on differentially expressed genes (DEG), and corresponding gene networks were constructed via Ingenuity Pathway Analysis (IPA). Through the network, we screened the potential “candidate targets,” such as ITGB1, FN1, CASP3, and ITGA5 and FABP1, ABCB1, FABP2, and SLC51B. These potential candidate targets were functionally related to immune responses, inflammation, and metabolism. Moreover, HQT significantly decreased serum levels of proinflammatory factors nitrogen monoxide (NO), proinflammatory cytokines interleukin- (IL-) 17, and prostaglandin E2 (PGE2). The degree of HE staining of colonic tissue was severe in the model group but reduced significantly in the HQT group. HQT exhibited protective effects against colon damage by inhibiting the inflammatory response.

1. Introduction

UC is a chronic nonspecific inflammatory disease involving the rectum and colon, characterized by mucosal T cell dysfunction, abnormal cytokine production, and cellular inflammation that lead to damage of the distal small intestine and the colonic mucosa [1, 2]. In severe cases, visible focal hemorrhages are apparent and the colonic mucosa becomes brittle and bleeds easily [3]. The importance of mucosal healing is now acknowledged in new guidelines, which recommend incorporating complete mucosal healing (along with symptom resolution) in all therapeutic studies as the primary endpoints of remission [4]. Most current therapies for UC include corticosteroids, glucocorticosteroids, aminosalicylates, and immunosuppressive agents. However, side effects are serious and the recrudescence rates of UC are high [5, 6]. Compared with chemical treatment, herbal medicine has the advantage of less toxic side effects, which is increasingly attracting researchers’ attention as a drug candidate for the treatment of such diseases [7].

HQT can be used to treat UC based on traditional Chinese medicine (TCM) theory. It has been widely used to treat gastrointestinal diseases, such as diarrhea, abdominal spasms, fever, headache, vomiting, nausea, extreme thirst, and subcardiac distention [8]. The major effective components of the four herbs are flavonoids (e.g., wogonin, baicalein, and oroxylin-A) and flavonoid glycosides (e.g., wogonoside, baicalin, and oroxylin-A-glucoside) [9–11], terpenoids, isoflavones, polysaccharides, and volatile oils. Phytochemical studies have shown that these compounds have a wide range of pharmacological characteristics [12–14], such as anti-inflammatory, antitussive, tumor suppressor, analgesic, and immunomodulatory properties [15–18]. Flavonoids from Radix Scutellariae are the major active ingredients from HQT.

With the soaring development of systems biology and polypharmacology, “network pharmacology” [19, 20] has shifted the paradigm of “one gene, one drug, one disease” to the “multi-component, network target” strategy [21], of which the key idea is in line with the holistic theory of TCM. To explore the pharmacological mechanisms of HQT in UC, we used genome-wide microarray detection and ingenuity pathway analysis (IPA) to identify 569 various genes (199 upregulated and 370 downregulated genes, , value < 0.05) in the model group compared to the normal group. Further, after data processing of the 569 genes, we found 162 genes (, value < 0.05) in the HQT group compared to the UC model group. We were then able to construct the network using the 162 the potential regulating genes of HQT by Cytoscape. Among them, we analyzed potential “candidate targets” (ITGB1, FN1, CASP3, and ITGA5 and FABP1, ABCB1, FABP2, and SLC51B) in terms of the “degree” ≥10 or | in the network. These potential candidate targets were functionally correlated with proliferation, inflammatory response, and metabolism, and these genes were revealed to be crucial components in the pathways of inflammation. Moreover, portions of gene expression values were validated by qRT-PCR. All in all, the integrative research combining microarray gene expression profiling, network analysis, and inflammation experiment reveals convincing evidence that HQT may alleviate UC via regulating immune or metabolism-related genes in a holistic way.

2. Materials and Methods

2.1. Animals

Male Wistar rats were obtained from the Laboratory Animal Center of the Academy of Military Medical Sciences; Production license No. was SCXK 2012–0004, weight 180–200 g. All rats were housed at . Animal experiment process was conducted in accordance with the ethical guidelines for local animal care and usage.

2.2. Preparations of HQT

Scutellaria baicalensis Georgi, Paeonia lactifloraPall, Glycyrrhiza uralensis Fisch, and Ziziphus jujuba Mill (weight ratio 3 : 2 : 2 : 3) were weighed and mixed. For the first decoction, the mixture was refluxed with water (1 : 10, ) for 1.5 h. The filtrates were collected; then, the residues in water (1 : 8, ) were boiled for an additional 1 h. Two batches of filtrates were gathered. Hereafter, the sample is dried under reduced pressure to obtain HQT extract. In addition, the major active chemical compounds of each herb containing in HQT were determined by liquid chromatography–tandem mass spectrometer (LC-MS).

2.3. UC Rat Model Construction and Grouping

TNBS colitis in rats was induced according to previously reported methods [22]. 24 h fasted Wistar rats were moderately anesthetized with pentobarbital then carefully inserted a 0.56 mm catheter into the colon with the tip 8 cm proximal to the anus. To break the intestinal epithelial barrier and induce colitis, the mixed reagent containing 0.25 ml of 50% ethanol and TNBS (100 mg/kg) was slowly administered into the lumen by the catheter fitted onto a 1 ml syringe. Rats in the control group were all administered an equal volume of saline. Animals were kept in an upside-down position for 2 min and returned to their cages.

For microarray analysis, male rats were divided into three groups randomly: a normal control group, a UC model group, and a group in which UC rats were treated with HQT formula (HQT group). Each group has 10 rats. The HQT group was administered orally at a daily dose of 20 g/kg for 10 consecutive days. Rats in the control and UC model groups received an equal volume of saline.

2.4. Gene Expression Data Analysis

On the 10th day, rats were killed under anesthesia. The colonic segments were collected from all three rat groups, freed of adherent adipose tissue, and washed with cool saline. Colonic segments were then frozen in liquid nitrogen immediately for further microarray detection. Whole transcriptome analysis was developed by Affymetrix GeneChip®Rat Gene 2.0 Array chips. Gene array data was uploaded to the IPA system. A cutoff was set to identify genes whose expression was significantly differentially regulated; these molecules were known as Network Eligible Molecules (NEMs). Networks of NEMs were then algorithmically formed based on their connectivity. The probability of the assignment was expressed by a value calculated using the right-tailed Fisher’s exact test. The level of statistical significance was set at . Subsequently, signal pathways with a value < 0.05 and were screened out and analyzed [23–28].

2.5. Serum Cytokine Detection

After a ten-day treatment, blood samples were collected by eyelid method and then centrifuged (3,000 rpm, 15 min) to get serum. Production of NO in serum was measured by Griess assay, and the levels of proinflammatory cytokines IL-17 and PGE₂ in serum were detected by ELISA kits according to the manufacturer’s specifications.

2.6. Histological Examination

After tissue collection, the tissue was embedded and sliced as usual. Then, 4 μm-thick tissue sections were prepared and stained with hematoxylin-eosin (HE) for histological studies. The extent of colonic lesions was compared based on ulcer size, inflammatory infiltration, and structural damage.

2.7. Reverse Transcription-PCR

SYBR Green 1 kit was applied under the manufacturer’s instructions, and the control housekeeping gene GAPDH was utilized as a reference gene. Then, 2 μg of total RNA was performed for reverse transcription using ABI High-Capacity cDNA reverse transcription Kits. Primer sequences of target genes are given in Table 1. Moreover, the mixtures were diluted tenfold and final reaction volumes of 20 μl were used for data analysis using BIO-RAD CFX Manager Version 3.0 software. Reactions were run in triplicate. The relative levels of target genes were calculated by the method [29].

2.8. Statistical Analysis

SPSS version 11.0 (SPSS Inc., Chicago, IL, USA) was used for statistical analysis. Values displayed represent the . Differences between mean values were analyzed by one-way analysis of ANOVA followed by LSD method. Significance of differences were accepted at values less than 0.05.

3. Results

3.1. Chemical Compounds of HQT

LC-MS was performed to identify the chemical compounds of each herb contained in HQT. The 10 main active ingredient chemical compositions were as follows: Baicalin (10.4149%), Wogonoside (2.4179%), Oroxylin-A-glucoside (0.8229%), Baicalein (0.6754%), Wogonin (0.2215%), Oroxylin-A (0.0967%), Liquiritin (0.0384%), Isoliquiritin apioside (0.0354%), Liquiritigenin (0.0280%), Isoliquiritoside (0.0865 μg/mL), and Isoliquiritigein (0.0049%) [30].

3.2. HQT Promoted Recovery of the UC Rats

The UC rats induced by TNBS showed a series of symptoms within 3 days, such as apathetic, drowsiness, dull hair, poor appetite, weight loss, diarrhea, and purulent stools. After prolonged HQT exposure, rats_, response to above symptoms were all improved to varying degrees. Their appetite was stimulated with decreased diarrhea. There was no significant improvement in symptoms in the model group rats.

3.3. HQT Decreases Serum Level of Inflammatory Cytokines

The levels of proinflammatory factors NO, IL-17, and PGE2 in the model group were increased remarkably in model group. In addition, levels of NO, IL-17, and PGE2 in the HQT group were clearly lower than those in the model group (Table 2).

3.4. Histological Study

The rats with TNBS-induced colitis in the mucosa of colons revealed inflammatory cell infiltration, loss or enlarged of goblet cells and epithelium, and edema. Part of the colon gland disappeared. The histologic sections of the HQT treatment group showed progressive restoration, improvement of intestinal ulcer, and reduction in infiltration of inflammatory cells and edema compared to that of the TNBS control group (Figure 1(b)). Considering internal organs as the executors of the physiological function of the body, the organ coefficient can be approximated to reflect the functional state of organs and disease. In this experiment, we observed that the weight of the spleen and kidney were, respectively, increased (, ) in the UC model group compared to the normal group; after treatment with HQT, the two parameters were both lower than those in UC model rats (). While the weight of the thymus was reduced () in the UC model group compared to the normal group, after the administration of HQT, the indicator was higher than those in UC model rats with no statistical significance (Figure 1(a)).

(a)

(b)

Figure 1

Effect of HQT on TNBS-induced ulcerative colitis rats. (a) Weights of the spleen and kidney were, respectively, increased in the UC model group compared to the normal group; after treatment with HQT, the two parameters were both lower than those in UC model rats. Weight of the thymus was reduced in the UC model group compared to the normal group; after treatment with HQT, the indicator was higher than that in UC model rats; (b) histological observations of the colon tissues in different groups (HE staining), (). A: normal control; B: UC model control; C: HQT 20 g·kg⁻¹. Data are represented as the . vs. normal control.

3.5. Heat Maps of Relative Expression Changes

By data processing and DEG screening, there were 199 upregulated and 370 downregulated genes in UC model rats compared to the normal rats. After HQT-treated, there were 114 upregulated and 90 downregulated genes in HQT rats compared to the UC model rats. In addition, unsupervised hierarchical clustering analysis (Figure 2(a)) of the dysregulated genes presented a good differentiation of normal and model samples, suggesting the successful construction of the UC model induced by TNBS. After HQT treatment, dysregulated genes tended to callback (Figure 2(b)).

(a)

(b)

3.6. HQT Alleviates UC Via Regulating Potential Key Genes

According to the IPA analysis, 162 genes (genes with a value of <0.05 and a , but with a after being HQT-treated) were viewed as significantly differentially expressed between the model and HQT groups. We then constructed the network utilizing interaction information among HQT-regulated genes (Figure 3). According to the pathway enrichment analysis and top diseases and functions based on IPA, the potential “candidate targets” (including ITGB1, FN1, and ITGA5) of the network were frequently involved Actin Cytoskeleton Signaling, Agranulocyte Adhesion and Diapedes, Integrin Signaling, NF-κB Activation by Viruses, FAK Signaling, PAK Signaling, and ERK/MAPK Signaling.

Figure 3

Analyses of interactions among 162 genes. Red and green nodes represent the differentially expressed genes identified in these networks. A red node denotes an upregulated gene, a green node denotes a downregulated gene, and a yellow node denotes a related inflammation gene. Genes in gray notes were not identified as differentially expressed in our experiment and were integrated into the computationally generated networks based on the evidence stored in the IPA database, indicating relevance to these networks.

3.7. Top Diseases and Functions

Based on potential gene interactions among the HQT-regulated genes, we applied the “Core” program of the IPA Software to identify biological and functional networks. The majority of these genes were classified into ten networks comprising functions (Table 3).

3.8. Experimental Verification of mRNA Levels

Real-time PCR experimental verification showed that the therapeutic effects of HQT on UC rats were consistent with findings based on microarray dates (Figure 4). Moreover, the results of real-time PCR analyses indicated that the mRNA levels of FN1 (), FABP1 (), FABP2 (), and SLC51B () were up- or downregulated with statistical significance while ITGB1 and ITGA5 were upregulated with no statistical significance. FABP1, FABP2, and SLC51B expressions in the colon tissues of UC rats were lower than those in normal rats, but could be effectively increased by HQT treatment, and the increased FN1 expression in UC model rats markedly reduced in the HQT group.

(a)

(b)

4. Discussion

From a systematic perspective and at a molecular level, we combined genome-wide microarray detection based on the colon tissues of UC rats and network target analysis to define HQT in the current study. Notably, a list of differentially expressed genes screened based on fold change and degrees in the interaction network were identified including FABP1, ABCB1, FABP2, SLC51B, ITGB1, FN, and ITGA5 (Tables 4 and 5). According to the top diseases and functions, these potential candidate targets were most significantly related to cellular growth and proliferation, cellular recombination, repair, and metabolism, all of which appear to be involved in UC progression. Moreover, the potential regulating genes were also found as crucial components in some pathways. These genes drew our attention, and we further analyzed them to explore pharmacological mechanisms of HQT acting in the setting of UC.

Integrins including ITGB1 are membrane receptors involved in cell adhesion and several processes, including immune response. ITGB1 is expressed during hypoxic conditions and can serve as an indicator of intestinal wound repair [31]. The increased levels of ITGB1 observed in the inflamed colon are consistent with previous studies that reported murine ITGB1 to be induced in a TNBS concentration-dependent manner [31]. Considering the ITGB1 gene, which functions in regulating cell adhesion, we analyzed HQT-mediated modulation of cell adhesion-related genes that may affect the attachment of one cell to another cell, suppress inflammation, and play a critical role in intestinal epithelium protection.

FN1, related to migration processes and cell adhesion, is poorly expressed in normal adult tissue but overexpressed in wound healing [32]. At the same time, our study here found FN1 to be elevated in the colon of the UC rat and HQT to effectively inhibit the expression of FN1. As another member of the integrin family, ITGA5 can combine with ITGB1 and form a heterodimer “Integrin α5β1.” Integrin α5β1 is a fibronectin (FN) receptor. Signal transduction could be triggered when Integrin α5β1 combined with FN1 to activate a variety of signaling molecules and participated in physiological processes such as cell adhesion and cell skeleton reconstruction [33]. According to canonical pathways, the genes ITGB1, ITGA5, and FN1 all associate with the pathways, such as agranulocyte adhesion and diapedes, NF-κB activation by viruses, FAK signaling, PAK signaling, and ERK/MAPK signaling. Activation of the ERK/MAPK and NF-κB pathways could trigger the expression of a series of proinflammatory factors such as IL-1β, NO, and PGE2. Destruction of the body’s immune homeostasis ultimately leads to UC [34, 35]. Our previous studies have shown that HQT can effectively inhibit the activation of ERK/MAPK and NF-κB pathways, thereby downregulating inflammatory mediators [36]. Experiments also show that HQT inhibits high expression of IL-17, NO, and PGE2. It is suggested that ITGB1, ITGA5, and FN1 may be involved in the regulation of inflammation by ERK/MAPK and NF-κB pathways.

Intestinal-type fatty acid-binding protein (I-FABP) has been used as a marker for the detection of acute intestinal injury [37]. The transport of fatty acids is believed to be performed by the FABP family. The result showed that FABP1 and FABP2 were downregulated in the inflamed UC mucosa. Our previous studies have shown that there was a large accumulation of fatty acids in UC rats [38]. The large accumulation of fatty acids can exacerbate the inflammatory response. The expression of FABP1 and FABP2b was upregulated in the HQT group. We raise the possibility that HQT enhanced the fatty acid transport and thus reduced the inflammatory response in UC rats.

Expression of SLC51B, which has a role in bile acid transport, was decreased in the UC group. The diminished uptake of bile acids into ileocytes is most likely responsible for the decreased expression of SlC51B in the ileum [39]. HQT can effectively increase SLC51B and FABP family gene expression, and we assume that the cells exist as a whole in response to disturbances. This leads to the activation of a range of pathways including functions such as metabolic regulation.

It is known that ABCB1 gene product P-glycoprotein (P-gp) exerts its barrier function by means of an efflux transport of drugs [40]. As ABCB1 mediates the efflux transport of xenobiotics and bacterial toxins, its downregulation in inflamed UC intestinal regions may be associated with compromised barrier function.

Analyzing the combination of organ coefficient, the thymus, as a central control organ of the immune system was decreased, and coefficients of the kidney and spleen, which are closely associated with energy metabolism, were increased in the UC model group as compared to the normal group. A previous study [41] suggests that persistence of disordered lipid metabolism can lead to kidney damage. We suspect that the changes of organ coefficients may be linked to metabolic disorder. After administration of HQT, compared to the UC model group, organ coefficients and the genes with the function of metabolism or transport of the HQT group rendered a callback trend, indicating that HQT could regulate dysfunctions of lipid and fatty acid metabolism, further improving the state of organ function.

In summary, this integrative study offers convincing evidence that HQT may alleviate UC via regulating the aforementioned potential key genes, which are involved in immune responses, inflammation, and metabolism. These novel findings may provide a novel and powerful mean to clarify HQT as an efficient drug candidate in therapy of UC.

Data Availability

The datasets used and analyzed during the current study are available from the corresponding author upon reasonable request.

Conflicts of Interest

The authors declare no conflict of interest.

Authors’ Contributions

Dunfang Wang and KaiFeng Shi made equal contributions to this work.

Acknowledgments

The work was supported by the National Natural Science Foundation of China (Nos. 81273662 and 81473592) and the Operational Expenses for Basic Research of the China Academy of Chinese Medical Sciences (No. ZZ2014020).

References

D. C. Baumgart and W. J. Sandborn, “Inflammatory bowel disease: clinical aspects and established and evolving therapies,” Lancet, vol. 369, no. 9573, pp. 1641–1657, 2007.
View at: Publisher Site | Google Scholar
H. Cheng, B. Xia, Q. S. Guo et al., “Sinomenine attenuates 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis in mice,” International Immunopharmacology, vol. 7, no. 5, pp. 604–611, 2007.
View at: Publisher Site | Google Scholar
G. R. Lichtenstein and P. Rutgeerts, “Importance of mucosal healing in ulcerative colitis,” Inflammatory Bowel Diseases, vol. 16, no. 2, pp. 338–346, 2010.
View at: Publisher Site | Google Scholar
G. D'Haens, W. J. Sandborn, B. G. Feagan et al., “A review of activity indices and efficacy end points for clinical trials of medical therapy in adults with ulcerative colitis,” Gastroenterology, vol. 132, no. 2, pp. 763–786, 2007.
View at: Publisher Site | Google Scholar
D. M. Abdallah and N. R. Ismael, “Resveratrol abrogates adhesion molecules and protects against TNBS-induced ulcerative colitis in rats,” Canadian Journal of Physiology and Pharmacology, vol. 89, no. 11, pp. 811–818, 2011.
View at: Publisher Site | Google Scholar
X. Liu and J. Wang, “Anti-inflammatory effects of iridoid glycosides fraction of Folium syringae leaves on TNBS-induced colitis in rats,” Journal of Ethnopharmacology, vol. 133, no. 2, pp. 780–787, 2011.
View at: Publisher Site | Google Scholar
Y. Jia, Q. Guan, Y. Jiang et al., “Amelioration of dextran sulphate sodium-induced colitis in mice by echinacoside-enriched extract of Cistanche tubulosa,” Phytotherapy Research, vol. 28, no. 1, pp. 110–119, 2014.
View at: Publisher Site | Google Scholar
S. H. Liu and Y. C. Cheng, “Old formula, new Rx: the journey of PHY906 as cancer adjuvant therapy,” Journal of Ethnopharmacology, vol. 140, no. 3, pp. 614–623, 2012.
View at: Publisher Site | Google Scholar
S. Ikemoto, K. Sugimura, N. Yoshida et al., “Antitumor effects of Scutellariae radix and its components baicalein, baicalin, and wogonin on bladder cancer cell lines,” Urology, vol. 55, no. 6, pp. 951–955, 2000.
View at: Publisher Site | Google Scholar
M. Li-Weber, “New therapeutic aspects of flavones: the anticancer properties of Scutellaria and its main active constituents Wogonin, Baicalein and Baicalin,” Cancer Treatment Reviews, vol. 35, no. 1, pp. 57–68, 2009.
View at: Publisher Site | Google Scholar
C. Li, G. Lin, and Z. Zuo, “Pharmacological effects and pharmacokinetics properties of Radix Scutellariae and its bioactive flavones,” Biopharmaceutics & Drug Disposition, vol. 32, no. 8, pp. 427–445, 2011.
View at: Publisher Site | Google Scholar
S. H. Wu, D. G. Wu, and Y. W. Chen, “Chemical constituents and bioactivities of plants from the genus Paeonia,” Chemistry & Biodiversity, vol. 7, no. 1, pp. 90–104, 2010.
View at: Publisher Site | Google Scholar
Q. Zhang and M. Ye, “Chemical analysis of the Chinese herbal medicine Gan-Cao (licorice),” Journal of Chromatography. A, vol. 1216, no. 11, pp. 1954–1969, 2009.
View at: Publisher Site | Google Scholar
S. Guo, Y.-P. Tang, J.-A. Duan, S.-L. Su, and D.-W. Qian, “Chemical constituents from the fruits of Ziziphus jujuba,” Chinese Journal of Natural Medicines, vol. 7, no. 2, pp. 115–118, 2009.
View at: Publisher Site | Google Scholar
D. Y. He and S. M. Dai, “Anti-inflammatory and immunomodulatory effects of Paeonia lactiflora Pall., a traditional Chinese herbal medicine,” Frontiers in Pharmacology, vol. 2, p. 10, 2011.
View at: Publisher Site | Google Scholar
J. Kamei, R. Nakamura, H. Ichiki, and M. Kubo, “Antitussive principles of Glycyrrhizae radix, a main component of the Kampo preparations Bakumondo-to (Mai-men-dong-tang),” European Journal of Pharmacology, vol. 469, no. 1-3, pp. 159–163, 2003.
View at: Publisher Site | Google Scholar
M. N. Asl and H. Hosseinzadeh, “Review of pharmacological effects of Glycyrrhiza sp. and its bioactive compounds,” Phytotherapy Research, vol. 22, no. 6, pp. 709–724, 2008.
View at: Publisher Site | Google Scholar
J. G. Jiang, X. J. Huang, J. Chen, and Q. S. Lin, “Comparison of the sedative and hypnotic effects of flavonoids, saponins, and polysaccharides extracted from semen Ziziphus jujube,” Natural Product Research, vol. 21, no. 4, pp. 310–320, 2007.
View at: Publisher Site | Google Scholar
A. L. Hopkins, “Network pharmacology,” Nature Biotechnology, vol. 25, no. 10, pp. 1110-1111, 2007.
View at: Publisher Site | Google Scholar
A. L. Hopkins, “Network pharmacology: the next paradigm in drug discovery,” Nature Chemical Biology, vol. 4, no. 11, pp. 682–690, 2008.
View at: Publisher Site | Google Scholar
S. Li, “Network target: a starting point for traditional Chinese medicine network pharmacology,” Zhongguo Zhong Yao Za Zhi, vol. 36, no. 15, pp. 2017–2020, 2011.
View at: Google Scholar
Y. W. Wang, H. H. Zhang, Y. L. Wang et al., “Effect of Huangqin Tang on the regulatory NF-κB p65 signal pathway in rats with ulcerative colitis,” Yao Xue Xue Bao, vol. 50, no. 1, pp. 21–27, 2015.
View at: Google Scholar
Y. Chen, C. Zhou, Y. Yu et al., “Variations in target gene expression and pathway profiles in the mouse hippocampus following treatment with different effective compounds for ischemia-reperfusion injury,” Naunyn-Schmiedeberg's Archives of Pharmacology, vol. 385, no. 8, pp. 797–806, 2012.
View at: Publisher Site | Google Scholar
Y. Chen, F. Meng, H. Fang et al., “Hierarchical profiles of signaling pathways and networks reveal two complementary pharmacological mechanisms,” CNS & Neurological Disorders Drug Targets, vol. 12, no. 6, pp. 882–893, 2013.
View at: Publisher Site | Google Scholar
J. Liu, C. X. Zhou, Z. J. Zhang, L. Y. Wang, Z. W. Jing, and Z. Wang, “Synergistic mechanism of gene expression and pathways between jasminoidin and ursodeoxycholic acid in treating focal cerebral ischemia–reperfusion injury,” CNS Neuroscience & Therapeutics, vol. 18, no. 8, pp. 674–682, 2012.
View at: Publisher Site | Google Scholar
Y. Y. Zhang, H. X. Li, Y. Y. Chen et al., “Convergent and divergent pathways decoding hierarchical additive mechanisms in treating cerebral ischemia-reperfusion injury,” CNS Neuroscience & Therapeutics, vol. 20, no. 3, pp. 253–263, 2014.
View at: Publisher Site | Google Scholar
Z. Wang, Z. W. Jing, C. X. Zhou et al., “Fusion of core pathways reveals a horizontal synergistic mechanism underlying combination therapy,” European Journal of Pharmacology, vol. 667, no. 1-3, pp. 278–286, 2011.
View at: Publisher Site | Google Scholar
Y. Zhang, M. Bai, B. Zhang et al., “Uncovering pharmacological mechanisms of Wu-tou decoction acting on rheumatoid arthritis through systems approaches: drug-target prediction, network analysis and experimental validation,” Scientific Reports, vol. 5, no. 1, 2015.
View at: Publisher Site | Google Scholar
M. Ballester, A. Castello, E. Ibanez, A. Sanchez, and J. M. Folch, “Real-time quantitative PCR-based system for determining transgene copy number in transgenic animals,” BioTechniques, vol. 37, no. 4, pp. 610–613, 2004.
View at: Publisher Site | Google Scholar
T. Li, S. Zhuang, Y. Wang et al., “Flavonoid profiling of a traditional Chinese medicine formula of Huangqin Tang using high performance liquid chromatography,” Acta Pharmaceutica Sinica B, vol. 6, no. 2, pp. 148–157, 2016.
View at: Publisher Site | Google Scholar
S. Keely, L. E. Glover, C. F. MacManus et al., “Selective induction of integrin beta1 by hypoxia-inducible factor: implications for wound healing,” The FASEB Journal, vol. 23, no. 5, pp. 1338–1346, 2009.
View at: Publisher Site | Google Scholar
K. Goossens, A. Van Soom, A. Van Zeveren, H. Favoreel, and L. J. Peelman, “Quantification of fibronectin 1 (FN1) splice variants, including two novel ones, and analysis of integrins as candidate FN1 receptors in bovine preimplantation embryos,” BMC Developmental Biology, vol. 9, no. 1, p. 1, 2009.
View at: Publisher Site | Google Scholar
S. M. Xu, The Role of Integrin-a5(ITGA5)in the Proliferation, Migration and Osteo-/Odontogenic Differentiation of Human Dental Pulp Stem Cells, Southern Medical University, 2013.
A. S. Baldwin Jr., “Series introduction: the transcription factor NF-kappaB and human disease,” Journal of Clinical Investigation, vol. 107, no. 1, pp. 3–6, 2001.
View at: Publisher Site | Google Scholar
X. L. Li, Y. Q. Cai, H. Qin, and Y. J. Wu, “Therapeutic effect and mechanism of proanthocyanidins from grape seeds in rats with TNBS-induced ulcerative colitis,” Canadian Journal of Physiology and Pharmacology, vol. 86, no. 12, pp. 841–849, 2008.
View at: Publisher Site | Google Scholar
H. H. Zhang, Anti-inflammatory effect and mechanism of Huangqin Tang[D], China Academy of Chinese Medical Sciences, 2014.
M. M. Pelsers, Z. Namiot, W. Kisielewski et al., “Intestinal-type and liver-type fatty acid-binding protein in the intestine. Tissue distribution and clinical utility,” Clinical Biochemistry, vol. 36, no. 7, pp. 529–535, 2003.
View at: Publisher Site | Google Scholar
D. F. Wang, Y. L. Wang, Y. W. Wang et al., “Effect of Huangqin Tang on serum metabolic profile in rats with colitis based on UHPLC-MS,” Acta Pharmaceutica Sinica, vol. 52, no. 8, pp. 1306–1312, 2017.
View at: Google Scholar
P. A. Dawson, M. Hubbert, J. Haywood et al., “The heteromeric organic solute transporter alpha-beta, Ostalpha-Ostbeta, is an ileal basolateral bile acid transporter,” The Journal of Biological Chemistry, vol. 280, no. 8, pp. 6960–6968, 2005.
View at: Publisher Site | Google Scholar
F. J. Sharom, “ABC multidrug transporters: structure, function and role in chemoresistance,” Pharmacogenomics, vol. 9, no. 1, pp. 105–127, 2008.
View at: Publisher Site | Google Scholar
P. Hu and C. X. Jing, “Dyslipidemia in the progression of renal disease,” Journal of Clinical Pediatric Dentistry, vol. 9, pp. 794–796, 2007.
View at: Google Scholar

Copyright

Copyright © 2020 Dunfang Wang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

560

Downloads

840

Citations