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Background Manganese (Mn) is one of the environmental factors of Parkinson's disease (PD), and long-term exposure to Mn can cause nerve damage. It is important to explore the common mechanism of neurotoxic effects of Mn and neurodegenerative diseases (NDD), especially PD, for early diagnosis of the disease. Objective To comprehensively analyze the core messenger RNA (mRNA)-microRNAs (miRNAs) co-expressed in frontal cortex of NDD patients and neuronal cells exposed to Mn via bioinformatics, and to reveal the potential common mechanism between Mn-induced neurotoxicity and NDD, especially PD. Methods Difference of the mRNAs from frontal cortex of NDD patients (GSE150696) and human neuroblastoma (SH-SY5Y) cells exposed to Mn were analyzed by R software; Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed on the overlapping differentially expressed genes (DEGs). The miRNAs were predicted using the miRNet database, mRNA-miRNA interactions were identified by the starBase and miRTarBase databases, and mRNA-miRNA regulatory networks were constructed with Cytoscape software. The core miRNAs associated with PD (GSE77667) were incorporated into Weighted Gene Co-Expression Network Analysis (WGCNA) and the mRNA-miRNA regulatory network was comparatively analyzed. Results A total of 34 overlapping DEGs were identified in the frontal cortical of NDD patients and the neuronal cells exposed to Mn, mainly enriched in interleukin-17 (IL-17) signaling pathway, cyclic adenosine monophosphate (cAMP) signaling pathway, and primary immunodeficiency. Based on the results of database prediction, 52 miRNAs with 71 pairs of interaction relationships were finally included to construct the miRNA-mRNA regulatory network. Six core miRNAs were screened by WGCNA: hsa-let-7i-5p, hsa-mir-155-5p, hsa-mir-219-2-3p, hsa-mir-221-3p, hsa-mir-485-3p, and hsa-mir-509-3-5p, among which hsa-let-7i-5p interacted with the target gene FBXW2 and hsa-mir-155-5p interacted with the target gene CCL2. The results of the KEGG analysis indicated that CCL2 was closely related to the IL-17 signaling pathway. Conclusion There are similar molecular regulatory mechanisms involved in the neurotoxicity of Mn and NDD, and the IL-17 signaling pathway may play a role in Mn-related NDD through CCL2 and hsa-mir-155-5p.
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Objective To explore the mechanism of SH?SY5Y mitochondrial dysfunction treated by manganese to find a new potential therapeutic target. Methods Transmission Electron Microscopy(TEM)to observe the morphology of mitochondria. Cell treated with 250μmol/L for periods of time(2 h, 4 h, 6 h)while mitochondrial membrane potential(MMP)and ROS can be detected by FCM and fluorescence microplate reader. Results After treating with MnCl2 in 6 h, TEM images showed early vacuoles, lamellar structures of SH?SY5Y cells. Then test the mitochondrial membrane potential and showed that MMP would be decreased gradually. Meanwhile, analysis showed that in comparison with control, treatment group had a higher ROS level respectively (P < 0.05). Conclusion MnCl2 can cause mitochondrial damage through a mechanism closely related to disrupt the MMP or generate abundant ROS.
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BACKGROUND:Drug eluting stents and endothelium stents for clinical treatment of vascular stenosis can lead to delayed endothelialization and restenosis. A rapamycin eluting stent combined with CD34 antibody can play a synergistic role to offset delayed endothelialization and intimal hyperplasia due to antiproliferative drugs, but it is stil in the pilot phase. OBJECTIVE:To observe the ability of rapamycin eluting stent combined with CD34 antibody to capture endothelial progenitor cels, and to observe the differentiation characteristics of the captured cels. METHODS:Scanning electron microscope and indirect immunofluorescence were used to observe the morphology and differentiation characteristics of captured endothelial progenitor cels. Under a fluorescence microscope, we observed the captured endothelial progenitor cels and the degree of endothelialization after implantation of the rapamycin eluting stent combined with CD34 antibody into rabbit ear vein. RESULTS AND CONCLUSION:Under the scanning electron microscope, fusiform-like cels with a diameter of 6-8 μm were captured by the composite stent, and 24 hours later, the cels became ful-shaped. The captured cels had the appearance characteristics of endothelial progenitor cels. Results from indirect immunofluorescence observation showed that there were a lot of red fluorescent spots on the coating which represented adherent cels positive for vascular endothelial growth factor receptor-2; the composite stent was largely covered with vascular endothelial cels at 24 hours after stent implantation, and fuly covered at 48 hours, but there was no abnormal cel cluster. These findings indicate that the rapamycin eluting stent combined with CD34 antibody can be specific to rapidly capture endothelial progenitor cels in the peripheral blood, and the stent can be completely covered with vascular endothelial cels at 48 hours after stent implantation, thereby achieving rapid endothelialization and promoting the repair of endothelial cels.
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Objective:To study the determination method of scutellarin in BREVISCAPINE INJECTION using HPLC.Methods:The waters ODS C 18 column (5?m,3.9?150mm) was used. The methanol water glacial acetic acid (40∶60∶1) was used as a mobile phase. The detection wavelength of scutellarin was set at 335nm and internal standard baicalin at 280nm. The resolution of scutellarin's and baicalin peaks was at least 5. Number of theoretical plates of column is no less than 1000 to scutellarin. Results:The recovery of additive sample is 99.8%. RSD is no more 2.0%.Conclusion: It is generally in agreement with external standard method.