Résumé
Objective To investigate the effect of cyclosporine A (CsA) on autophagylysosomal pathway in tubular epithelial cells.Methods Human renal tubular epithelial cell line (HK-2 cell) was treated with different concentrations (3,5 and 10 μmol/L) of CsA for 24 h.Then the viability and apoptosis of cells were measured by MTT assay or AnnexinV-PI staining followed by flow cytometry analysis,respectively.Autophagy-related protein LC3-Ⅱ and p62 were detected by immunofluorescence assay.Autophagic flux was analyzed in HK-2 cells transfected with a tandem mRFP-GFP fluorescent-tagged LC3 (ffLC3) plasmid by laser confocal microscope.The lysosomal degradation was evaluated by DQ-ovalbumin staining followed by flow cytometry analysis.Results The viability of HK-2 cells was significantly decreased with CsA stimulation when compared with control group (P ＜ 0.01),but the number of apoptotic cells was markedly increased by CsA treatment (P ＜ 0.05).Compared with the control group,different doses of CsA dramatically increased the expressions of LC3-Ⅱ (P ＜ 0.01) and p62 (P ＜ 0.05) in HK-2 cells.Moreover,HK-2 cells treated with CsA displayed a significant increase in autophagosomes but a marked decrease in autolysosomes.In HK-2 cells,exposured to CsA caused a decrease in lysosomal degradation by DQ-ovalbumin staining when compared with control group (P ＜ 0.01).Conclusion Blockade of autophagy via disrupting lysosome degradation may represent a novel mechanism of CsA-induced tubular epithelial cells injury.
Résumé
Objective To seek the ancients’ method for acupuncture treatment of epigastric pain and explore the ancients’ regularities in point selection for acupuncture treatment of this disease to provide a reference and basis for guidance of modern clinical treatment. Method Search words related to epigastric pain were established. A database was set up. Frequency analysis and data mining technique-associated rules were used to investigate the use frequencies of different acupoints and meridians, and regularities in acupoint combination in ancient acupuncture treatment of epigastric pain. Result In ancient acupuncture treatment of epigastric pain, the points more frequently used and more accepted were Zhongwan (CV12), Zusanli (ST36), Neiguan (PC6), Shangwan (CV13), Geshu (BL17), Gongsun (SP4) and Jianli (CV11). Meridian points were the main ones. The points of meridians going through the epigastrium were often selected. Acupoint combination was mainly based on syndrome differentiation-based point selection plus symptomatic point selection.