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Objective To determine the effect of Salvia przewalskii extract of total phenolic acids (SPE) on puromycin aminonucleoside (PAN)-induced oxidative stress in podocytes of rats in vivo and the effect of SPE on PAN-induced oxidative stress in podocytes of mice in vitro. Methods (1) Nephropathy rat model was established by PAN and was given intervention with SPE and tacrolimus. The renal tissue samples were obtained for WT1 staining to calculate the number of podocytes on the 5th# 10th# 15th and 21st day. The intensities of 8-hydroxy-27-deoxyguanine (8-OHdG) were evaluated by immunofluorescence. (2)The podocytes of mice were exposed to PAN for 24 h in vitro#, and then SPE# salvianolic acid B (SalB) # rosmarinic acid (RA) or tacrolimus were added for 6# 12# 24# and 48 h culture. Then the cytoskeleton distribution of podocytes, indicated by F-actin# was observed by fluorescence microscopy, and the intracellular reactive oxygen species (ROS) production was measured by flow cytometry. Results (1) Decrease of podocytes per glomerular volume as measured by counting WT1-positive cells was started on day 5 in each group except normal control (NC) group# and on day 15 glomerular podocytes in PAN group was significantly less than that in the NC group ([14. 4 + 0. 7]/glomerular volume vs [37. 2 + 1. 5]/glomerular volume# P<0. 05). The numbers of glomerular podocytes in SPE group and positive group (tacrolimus group) were more than that in PAN group at all time points. The glomerular podocyte count of high-dose SPE group was similar to that of positive group on day 15 ([21. 7 + 1. 0]/glomerular volume vs [23. 6 + 1. 2]/glomerular volume# P<0. 05). After injection of PAN# 8-OHdG intensities were increased in each group except normal control group on day 5; and the intensities peaked on day 10 and then began to decrease# but still higher than that of the normal control group on day 15. The intensities of 8-OHdG in renal tissue was decreased after intervention# and those of the tacrolimus and high-dose SPE groups were similar. (2) In vitro study found that F-actin of podocytes was almost completely disrupted 24 h after PAN treatment# with disrupted filamentous structure. After the treatment with tacrolimus, SPE, SalB and RA# the PAN induced injury of podocytes was lessened# with reappeared polarity distribution of intracellular microfilaments. Compared with NC group# the ROS production in podocytes was significantly increased in PAN group (P<0. 05). After treatment of podocyte with drugs# the ROS production was decreased. The cellular ROS production of positive control group was similar to those in tacrolimus group, low-dose SPE group, high-dose SalB group and RA group at 24 h. Compared with RA,SalB had a better efficacy in reducing ROS# and the reducing effect had a positive relation with drug dose. Conclusion Our study suggests that SPE can protect podocytes from PAN-induced oxidant stress in vivo and in vitro.
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Objective:To examine the content changes of neochlorogenic acid, chlorogenic acid, 1, 5- dicaffeoylquinic acid and total phenolic acids in the water extract from raw and fried Xanthii Fructus. Methods:The stir-frying method was used to process fried Xanthii Fructus from different habitats. The contents of neochlorogenic acid, chlorogenic acid and 1,5-dicaffeoylquinic acid in the wa-ter extract were determined by HPLC, the total phenolic acids content was determined by UV. Results:The contents of neochlorogenic acid, chlorogenic acid, 1,5-dicaffeoylquinic acid and total phenolic acids in the water extract from fried Xanthii Fructus were all in-creased. Conclusion:Fried Xanthii Fructus can increase the contents of effective ingredients in the decoction resulting in the enhanc-ment of clinical curative effect.
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Objective To screen extraction process for Rosae Laevigatae Fructus.Methods With the content of total saponins and total phenolic acids from Rosae Laevigatae Fructus as the evaluation indexes, ethanol concentration, extraction time, solid-liquid ratio and extraction times as investigation factors, the test data were analyzed by Design-Expert8.0.5 software, and a multiple quadratic regression equation was set up to screen extraction process parameters of Rosae Laevigatae Fructus.Results The best extraction process parameters of Rosae Laevigatae Fructus were as follows:ethanol concentration was 62.79%;solid-liquid ratio was 13.12:1;the extraction time was 137.93 min;extraction times were 1.85 times. Considering the convenience for actual operation and the practicability of industrial production, the extraction process parameters should be ethanol concentration of 60%, revised solid-liquid ratio of 13:1, 2 hours' extracting time, and 2 extraction times.Conclusion Final selected parameters provide the basis for the extraction process of Rosae Laevigatae Fructus.
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Objective: To determine the contents of phenols in Taohong Siwu Decoction (TSD) by ISC-HPLC. Methods: The ISC-HPLC was used to determine the contents of hydroxysafflor yellow A, ferulic acid, gallic acid, protocatechuic acid, and chlorogenic acid. Spectrophotometric method was used to determine the contents of total phenolic acids. Results: The contents of hydroxysafflor yellow A, ferulic acid, gallic acid, protocatechuic acid, and chlorogenic acid in TSD had good linear relationships with the peak area in the ranges of 5.00-80.00, 1.25-20.00, 0.60-9.60, 0.90-14.40, and 1.00-16.00 μg/mL, respectively, and the average recoveries (n = 9) were 95.82%, 94.98%, 96.52%, 97.16%, and 98.14%, respectively. Conclusion: ISC-HPLC method is simple, rapid, accurate, and reproducible, and can be used for the quality control of TSD.
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Objective: To optimize the processing method and technology for the fruits of Capparis spinosa. Methods: The determination methods for the effective fractions (total phenolic acids and total glucosinolates) and the stimulating ingredients in the fruits of C. spinosa were established by UV and GC, respectively. Four processing methods (steaming, decocting, baking, and stir-frying) were optimized with the contents of effective fractions and the stimulating ingredients as indexes. The processing technology was optimized by the single factor and orthogonal design, and the data treated by the desirability functions. Results: The best processing method for the fruits of C. spinosa was stir-frying, and the best processing technology was suggested as follows, stir-fried for 15 min at 80°C with the drug powder of 40-50 meshes. Conclusion: The processed fruits of C. spinosa could ensure the effective components and reduce their stimulation.