Mechanisms underlying the therapeutic effects of Semen cuscutae in treating recurrent spontaneous abortion based on network pharmacology and molecular docking.

Background: This paper aims to analyse the active components of Semen cuscutae (SC) by network pharmacology and screen the most stable compounds with tumour necrosis factor-alpha (TNF-α) by molecular docking to explore the mechanisms of SC treatment of recurrent spontaneous abortion (RSA) and provide a theoretical basis for drug development. Methods: The active compounds of SC and the potential inflammatory targets of RSA were obtained from the Traditional Chinese Medicine Systems Pharmacology database and GeneCards, respectively. The RSA-SC target gene interaction network was obtained and visualized using the STRING database and Cytoscape software. GO and KEGG pathway enrichment analyses were obtained from DAVID to further explore the RSA mechanism and therapeutic effects of SC. Interactions between TNF-α and drugs were analysed by molecular docking. Treatment of human trophoblast cells with sesamin and TNF-α was carried out to detect their proliferative and apoptotic abilities, and WB assay was carried out to detect EGFR, PTGS2, and CASP3 protein expression. Results: Ten compounds and 128 target genes were screened from SC, of which 79 overlapped with RSA target inflammatory genes, which were considered potential therapeutic targets. Network pharmacological analysis showed that sesamin, matrine, matrol, and other SC compounds had a good correlation with the inflammatory target genes of RSA. Related genes included PGR, PTGS1, PTGS2, TGFB1, and CHRNA7. Several signalling pathways are involved in the pathogenesis of RSA, such as the TNF-α signalling pathway, HIF-1 signalling pathway, oestrogen signalling pathway, proteoglycans in cancer cells, and FoxO signalling pathway. Molecular docking results suggested that sesamin was the most suitable natural tumour necrosis factor inhibitor (TNFi). Sesamin can promote proliferation and inhibit apoptosis in human trophoblasts by downregulating EGFR and CASP3 expression and upregulating PTGS2 expression. Conclusion: Our findings play an important role and basis for further research into the molecular mechanism of SC treatment of RSA and drug development of TNFi.

Generation and characterization of iPS cell line (CTGUi001-A) from skin fibroblasts of a patient with Fabry disease.

We have generated an iPSCs line (CTGUi001-A) from dermal fibroblasts of a 16-year-old male Fabry disease patient with a novel GLA gene mutation (c.156C > A) using Sendai virus encoding the four Yamanaka factors OCT4, SOX2, KLF4, and c-MYC. The CTGUi001-A iPSC line displayed typical embryonic stem cell-like morphology, carried the GLA gene mutation, expressed several pluripotent stem cell makers, retained normal karyotype (46, XY) and was capable of forming teratomas containing three germ layers.

miR-146a-5p-mediated suppression on trophoblast cell progression and epithelial-mesenchymal transition in preeclampsia

OBJECTIVE: This study aims to identify the effect of miR-146a-5p on trophoblast cell invasion as well as the mechanism in preeclampsia (PE). METHODS: Expression levels of miR-146a-5p and Wnt2 in preeclamptic and normal placentae were quantified. Trophoblast cells (HTR-8) were separately transfected with miR-146a-5p mimic, miR-146a-5p inhibitor, pcDNA3.1-Wnt2 or sh-Wnt2, and then the expression levels of miR-146a-5p, Wnt2, and epithelial-mesenchymal transition (EMT)-related proteins (Vimentin, N-cadherin and E-cadherin) were measured. Moreover, the proliferative, migratory and invasive capacities of trophoblast cells were detected, respectively. Dual luciferase reporter assay determined the binding of miR-146a-5p and Wnt2. RESULTS: Compared with normal placental tissues, the placentae from PE patients showed higher miR-146a-5p expression and lower Wnt2 expression. Transfection of miR-146a-5p inhibitor or pcDNA3.1-Wnt2 exerted pro-migratory and pro-invasive effects on HTR-8 cells and encouraged EMT in HTR-8 cells; transfection with miR-146a-5p mimic or sh-Wnt2 weakened the proliferative, migratory and invasive capacities as well as reduced EMT process of HTR-8 cells. Moreover, Wnt2 overexpression could partially counteract the suppressive effects of miR-146a-5p overexpression on the progression and EMT of HTR-8 cells. CONCLUSION: miR-146a-5p mediates trophoblast cell proliferation and invasion through regulating Wnt2 expression.

The rational discovery of multipurpose inhibitors of the ornithine decarboxylase.

Metabolic reprograming is a hallmark of cancer, and the polyamine metabolic network is dysregulated in many cancers. Ornithine decarboxylase (ODC) is a rate-limiting enzyme for polyamine synthesis in the polyamine metabolic network. In many cancer cells, ODC is over-expressed, so this enzyme has been an attracting anti-cancer drug target. In the catalysis axis (pathway), ODC converts ornithine to putrescine. Meanwhile, ODC's activity is regulated by protein-protein interactions (PPIs), including the ODC-OAZ1-AZIN1 PPI axis and its monomer-dimer equilibrium. Previous studies showed that when ODC's activity is inhibited, the PPIs might counteract the inhibition efficiency. Therefore, we proposed that multipurpose inhibitors that can simultaneously inhibit ODC's activity and perturb the PPIs would be very valuable as drug candidates and molecular tools. To discover multipurpose ODC inhibitors, we established a computational pipeline by combining positive screening and negative screening. We used this pipeline for the forward screening of multipurpose ligands that might inhibit ODC's activity, block ODC-OAZ1 interaction and enhance ODC non-functional dimerization. With a combination of different experimental assays, we identified three multipurpose ODC inhibitors. At last, we showed that one of these inhibitors is a promising drug candidate. This work demonstrated that our computational pipeline is useful for discovering multipurpose ODC inhibitors, and multipurpose inhibitors would be very valuable. Similar with ODC, there are a lot of proteins in human proteome that act as both enzymes and PPI components. Therefore, this work is not only presenting new molecular tools for polyamine study, but also providing potential insights and protocols for discovering multipurpose inhibitors to target more important protein targets.

[Influence of Rhubarb extract on the expression of NGAL and IL-18 in the kidney of rabbits after CPR].

OBJECTIVE: To study the preventive effect and mechanism of Rhubarb extract on renal injury after cardiopulmonary resuscitation (CPR) in rabbits with cardiac arrest (CA). METHODS: Twenty-four male Japanese big-ear rabbits were divided into sham operation group, CPR model group and Rhubarb pretreatment group by random number table method, with 8 rabbits in each group. The rabbits in Rhubarb pretreatment group were treated with Rhubarb extract 5 mL×kg-1×d-1 for 7 days; and those in sham operation group and CPR model group were fed with 0.9% normal saline 10 mL/d for 7 days. After 7 days, ventricular fibrillation (VF) was produced in CPR model group and Rhubarb pretreatment group by 50 V alternating currents stimulation through bottom of the heart leads to the apex to prepare CPR model. The rabbits of the CPR model group and Rhubarb pretreatment group were sacrificed at 2 hours after successful resuscitation, and the animals in the sham operated group were sacrificed directly after anesthesia. The levers of blood urea nitrogen (BUN) and creatinine (Cr) in serum were examined by automatic biochemical analyzer. The positive expression area of neutrophil gelatinase-associated lipocalin (NGAL) and interleukin-18 (IL-18) in kidney were examined by immunohistochemistry. RESULTS: Compared with the sham operation group, the levels of BUN and Cr were significantly increased in the CPR model group and Rhubarb pretreatment group [BUN (mmol/L): 15.53±3.90, 10.51±3.16 vs. 7.03±2.23, Cr (µmol/L): 137.20±12.23, 86.80±7.67 vs. 66.39±5.47, both P < 0.05]. Compared with the CPR model group, the levels of BUN and Cr were significantly decreased in the Rhubarb pretreatment group [BUN (mmol/L): 10.51±3.16 vs. 15.53±3.90, Cr (µmol/L): 86.80±7.67 vs. 137.20±12.23, both P < 0.05]. Immunohistochemical staining showed that the expression of NGAL and IL-18 mainly existed in glomerular and tubular cells in patina. Compared with the sham operation group, the positive expression areas of NGAL and IL-18 in kidney were significantly increased in the CPR model group and Rhubarb pretreatment group [NGAL (µm2): 208.26±7.58, 136.74±5.33 vs. 98.93±7.83, IL-18 (µm2): 256.48±4.64, 113.22±6.98 vs. 77.06±6.47, all P < 0.05]. Compared with the CPR model group, the positive expression areas of NGAL and IL-18 were significantly decreased in the Rhubarb pretreatment group [NGAL (µm2): 136.74±5.33 vs. 208.26±7.58, IL-18 (µm2): 113.22±6.98 vs. 256.48±4.64, both P < 0.05]. CONCLUSIONS: CA can lead to acute kidney injury (AKI). Rhubarb extract can reduce the expression of NGAL and IL-18 in kidney of rabbits after CPR, and protect the kidney after CPR.