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Objective::To explore the active components, potential targets and signaling pathways of Rhei Radix et Rhizoma in the treatment of renal fibrosis based on the network pharmacology method, and then to verify the target genes in vitro. Method::Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicine Integrated Database (TCMID) were retrieved to obtain the main active ingredients of Rhei Radix et Rhizoma. The potential anti-renal fibrosis targets of Rhei Radix et Rhizoma were predicted by similarity ensemble approach (SEA), Swiss Institute of Bioinformatics (SIB) and GeneCards Database. Target protein-protein interaction (PPI) network was constructed by using String Version 10.5 database. David 6.8 software was used for gene ontology (GO) enrichment analysis and the Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of the key targets. Cytoscape Version 3.6.0 software was used for visualized analysis of PPI network, active ingredient-key target network and the ingredient-target-signal pathway network. In combination with Malachards database, the signal pathways with high correlation with renal fibrosis were screened. Then, cell experiments were used for verification: HK-2 cells were selected to establish fibrosis model by transforming growth factor-β1 (TGF-β1) stimulation. The cells were treated with rhein for 48 hours. Western blot assay was used to detect the protein expression level of hypoxia inducible factor-1 α (HIF-1 α), vascular endothelial growth factor (VEGF), and platelet-derived growth factor receptor-α (PDGFR-α). Protein expression levels of E-cadherin and α smooth muscle actin (α-SMA) were detected by immunofluorescence. Apoptosis was detected with flow cytometry. Result::Totally 17 active ingredients of Rhei Radix et Rhizoma and 424 targets of anti-renal fibrosis effect were screened out, including five key targets: protein kinase B(Akt)1, mitogen activated protein kinases 3(MAPK3), epidermal growth factor receptor(EGFR), interleukin(IL)-6 and VEGFA in turn. The biological process of GO enrichment mainly involved signal transduction, cell proliferation and apoptotic process. The results of KEGG pathway enrichment showed that phosphatidylinositol 3-kinase(PI3K)/Akt, HIF-1, VEGF, and forkhead transcription factor (FoxO) pathways were related to the anti-renal fibrosis mechanism of Rhei Radix et Rhizoma. Results of the in vitro experiment proved that rhein could inhibit the expression of E-cadherin, α-SMA, HIF-1α, VEGF and PDGFR-α. In addition, rhein inhibited apoptosis induced by TGF-β1 in HK-2 cells. Part of the prediction results of network pharmacology were verified. Conclusion::This study reflects the multi-component, multi-target and multi-pathway mechanism characteristics of Rhei Radix et Rhizoma. The mechanisms of its anti-renal fibrosis effects may be related to inhibiting HIF-1 α / VEGF /PDGFR-α signaling pathway, apoptosis and epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells.
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Objective:To explore the effect of different effective parts of Taohe Chengqitang on the synthesis and degradation of extracellular matrix in human kideny-2（HK-2） cells induced by transforming growth factor-β1（TGF-β1）. Method:Petroleum ether extract, ethyl acetate extract, n-butanol extract, raffinate and polysaccharide extract, mirabilite extract were extracted with 70% ethanol by systematic solvent method. The HK-2 cell fibrosis model induced by TGF-β1 was built to intervene the cells in different parts of Taohe Chengqitang with different concentrations (0, 50, 100, 200, 400, 800 mg·L-1). Enzyme-linked immunosorbent assay（ELISA）kit assay was used to detect collagen（Col）-Ⅰα1 and fibronectin （FN）in supernatant to screen out the main active parts. Cell counting kit-8 （CCK-8）method was used to determine the best concentration of intervention site of bioactive components. Western blot analysis was used to detect the expression levels of Col-Ⅰ, Col-Ⅲ, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase inhibitor2 (TIMP2), and connective tissue growth factor (CTGF). Immunofluorescence assay was used to detect the expression of α-smooth muscle actin（α-SMA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） analysis was used to detect the mRNA expression of plasminogen activator inhibitor-1（PAI-1）. Result:ELISA kit assay demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract significantly reduced the Col-Ⅰα1 and FN content at the concentrations of 200 and 400 mg·L-1 (P<0.05, P<0.01). CCK-8 assay showed that the cells viability was significantly inhibited with drug intervention at the concentrations of 400 and 800 mg·L-1 (P<0.01). Western blot demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract decreased the expression levels of Col-Ⅰ, Col-Ⅲ, TIMP2 and CTGF in HK-2 cells induced by TGF-β1, and increased the expression of MMP-2 (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of immunofluorescence showed that ethyl acetate extract, n-butanol extract and chloroform extract could inhibit the expression of α-SMA (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of Real-time PCR showed that ethyl acetate extract and chloroform extract inhibited mRNA expression of PAI-1 (P<0.05), with more significant effect in n-butanol extract (P<0.01). Conclusion:The extracts of ethyl acetate, n-butanol and chloroform are the active parts of Taohe Chengqitang with the anti-renal fibrosis effect, with n-butanol extract as the most active part. The mechanism on anti-renal fibrosis may be related to its regulation of extracellular matrix (ECM) synthesis and degradation.
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Objective: To investigate the influence of ECG-triggered technique on imaging quality and parameters in diffusion weighted intravoxel incoherent motion imaging (IVIM DWI). Methods: Twenty young healthy volunteers underwent liver IVIM DWI examination were enrolled, the images were obtained with ECG-triggered and free breath, and SNR, ADC and parameters of IVIM (true-diffusion coefficient [D], pseudo-diffusion coefficient [D*], perfusion fraction [f]) of the left and right lobe of liver were acquired. The scores of liver imaging quality was analyzed with Wilcoxon sign rank sum test. The SNR, ADC, D, D* and f values were analyzed with paired-samples t test. The consistency of ADC, D, D*, and f were assessed with the Bland-Altman method. Results: The scores of liver imaging quality of ECG-triggered were significantly higher than those of free breath (all P<0.01). Under different b values, SNR of left lobe of liver with ECG-triggered were higher than those of the free breath (all P<0.01). The values of ADC and f of left lobe of liver with ECG-triggered were statistically lower than those of free breath (both P<0.05), but the values of D were higher than that of free breath (P<0.05). The consistency of left lobe of liver with ECG-triggered was better than that with free breath. Conclusion: IVIM DWI combined with ECG-triggered can be used to obtain better imaging quality and effectively overcome the interference of heart beating on parameters measuring in left lobe of liver.
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The original version of this article unfortunately contained a mistake. The presentation of the affiliation number was incorrect. The corrected one is given below.Zhong-zhu AI () 1†.
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Objective To evaluate the reproducibility of normal liver ADC measurements by different respiratory motion compensation techniques. Methods A total of 31 young healthy volunteers who are 20 to 40 years old without any hepatic diseases were selected to research. Each volunteer underwent liver DWI twice in 24 hours with the same parameters and location. The imaging was performed with free-breath(FB), breathhold(BH), rspiratory-triggered(RT)and navigator-triggered(NT)techniques, and the ADC values of the left hepatic lobe and right hepatic lobe (upper, middle and lower) was acquired with two scans. Analysis the the ADC values of various anatomic locations of liver with two-way analysis of variance of randomized block design. Reproducibility of ADCs was assessed with the Bland-Altman method. Analysis of variance and paired-sample t test was used to assess ADCs from both right and left liver lobe among the four techniques. Result The ADC values acquired from the four techniques were significant differences (P<0.01),and the ADC values of the right lobe were less than the left lobe's(P<0.01). It showed a trend to decrease moving from superior to inferior levels in both left and right lobes, and the ADC values among The middle and lower were significant differences (P<0.01). The limit of agreement of ADC of twice imaging among the four techniques were as follow: the right lobe was less than the left lobes, and the breathhold was less the others. As the result, reproducibility in the right liver lobe was better to that in the left and the reproducibility with breathhold was better than the other respiratory motion compensation techniques. Conclusions Both anatomic location and DWI technique influence the liver ADC measurements and their reproducibility. The reproducibility of BH is the best.