The capacity of reverse cholesterol transport (RCT) of lipid loaded cells is critical of atherosclerosis(AS) development.Autophagy is an important regulatory mechanism promoting RCT.But the exact effects of autophagy activation or inhibition on RCT remain unclear in AS.So in this review we suggest that partly inhibition of autophagy or induction of lysosomal biogenesis improves the autophagy capacity of lipid loaded cells.

OBJECTIVETo study the expression of Toll-like receptor 4 (TLR4) in renal tubular epithelial cells exposed to high glucose and the effect of spironolactone on the TLR4 expression.

METHODSIn vitro renal tubular epithelial cells (NRK-52E) were randomly exposed to DMEM culture solution with low glucose (5 mmol /L), high glucose (25 mmol/L) or 10(-7) mol/L spironolactone plus 25 mmol/L glucose. Immunohistochemistry, RT-PCR and Western blot were used to determine TLR4 protein and mRNA expression. The levels of IL-6 and TNF-alpha in the cell culture supernatant were determined using ELISA.

RESULTSThe expression of TLR4 mRNA in the high glucose group began to increase 6 hrs and remained at a higher level up to 24 hrs after exposure as compared with the low glucose group. The TLR4 mRNA expression in the spironolactone treatment group was significantly lower than that in the high glucose group, although it was higher than that in the low glucose group between 6 and 24 hrs after exposure. TLR4 protein expression increased significantly in the high glucose group 24 and 48 hrs after exposure compared with that in the low glucose group. The TLR4 protein expression in the spironolactone treatment group was lower than that in the high glucose group, but higher than that in the low glucose group. IL-6 and TNF-alpha expression in the supernatant from the NRK-52E cells in the high glucose groups increased significantly as compared with the low glucose group. The spironolactone treatment group had significantly reduced IL-6 and TNF-alpha expression compared with the high glucose group.

CONCLUSIONSHigh glucose triggers an increase in the expression of TLR4 and inflammatory factors in NRK-52E cells. TLR4 may participate in the progress of diabetic nephropathy. Spironolactone can reduce expression of TLR4 and inflammatory factors, which might be attributed to one of the mechanisms of protection by spironolactone against diabetic nephropathy.

Cells, Cultured ; Diabetic Nephropathies ; etiology ; Epithelial Cells ; metabolism ; Humans ; Hyperglycemia ; metabolism ; Immunohistochemistry ; Interleukin-6 ; analysis ; Kidney Tubules ; metabolism ; RNA, Messenger ; analysis ; Spironolactone ; pharmacology ; Toll-Like Receptor 4 ; analysis ; genetics ; physiology ; Tumor Necrosis Factor-alpha ; analysis

To establish the suitable modern drying processing parameters for Scrophulariae Radix (SR). With reference to the traditional drying processing method of SR and the characteristics of modern drying equipment, the drying process for SR was simulated as the following three stages: temperature-controlled drying-tempering-temperature-controlled drying. Eighteen batches of SR samples were obtained by the drying methods after the orthogonal design experiment with seven factors namely temperature, wind speed, and target moisture for the first stage, tempering time and temperature, as well as temperature and wind speed for the second stage. UPLC-TQ-MS was applied for determination of nine target compounds including catalpol, harpagide, verbascoside, ferulic acid, angroside-C, aucubin, harpagoside, cinnamic acid and ursolic acid in those dried samples and another 19 batches of SR samples collected from genuine producing area. Principal Component Analysis (PCA) was performed, and total energy consumption was also taken into consideration for analysis and evaluation. Results showed that the optimal drying processing method for SR was as follows: drying temperature of 60 ℃, drying wind speed of 50 Hz, and 50% for target moisture in the first stage; 24 h for tempering time and temperature of 20 ℃ in the second stage; drying temperature of 60 ℃, and drying wind speed of 30 Hz in the third stage. The medicinal materials with optimized modern drying processing method were extremely similar to those collected from genuine producing area in the aspect of both external properties and target compounds, and they were in line with the 2015 version of "Chinese Pharmacopoeia" requirements. In addition, they could help to shorten the drying time and increase the efficiency of primary processing, and thus promote the normalization and standardization of primary drying processing for SR.