ABSTRACT
Objective To establish a quality control method for monitoring the blood concentrations of cyclosporin A and tacrolimus by HPLC-MS/MS,and to evaluate the quality control samples using the Westgard multi-rule theory.Methods HPLC-MS/MS was used to determine the concentration of cyclosporin A and tacrolimus in human whole blood.The quality control samples of low,medium and high concentration levels in the therapeutic drug monitoring process were statistically analyzed,Levery-Jennings and Z-score quality control charts were drawn,and the Westgard multi-rule theory was applied for in-house quality control evaluation.Results The established method was fully validated with linear ranges of 10.40-1 040.00 ng·mL-1 and 0.50-49.50 ng·mL-1,the quantification limits were 10.40 and 0.50 ng·mL-1,respectively.The extraction recoveries were 108.61％-113.24％and 101.99％-109.37％,respectively.The matrix factors normalized by internal standard were 106.68％-111.27％and 95.70％-97.81％for cyclosporin A and tacrolimus,respectively.The intra-day and inter-day accuracy and precision were less than 15.0％.Other parameters were also validated and met the acceptance criteria.Levery-Jennings and Z-score quality control charts showed that there were 4 warnings(violation of the 12s rule)in the results of the 26 groups of quality control samples in the third quarter of 2022,and no phenomenon was observed to be out of control.Conclusion The established in-house quality control system for therapeutic drug monitoring of cyclosporin A and tacrolimus can effectively ensure the accuracy of blood drug concentration detection.
ABSTRACT
High-fat diet (HFD) feeding is a risk factor for kidney damage with limited treatment options. This study explored the effect of total glucosides of paeony (TGP) for treating obesity-related kidney damage. C57BL/6 mice fed with HFD were used for in vivo experiments. Body weight, lipid and renal function indicators were measured. Hematoxylin and eosin, Masson, Sirius Red and F4/80 immunohistochemical staining were performed. Fibrosis levels, inflammatory factors, MyD88, IκB and p-Jun NH2-terminal kinase (JNK) were detected. SV40 cells were incubated with palmitic acid, transfected with siRNA MyD88 and/or treated with TGP. The expression of MyD88, IκB, p-JNK, fibrosis and inflammatory factors were detected. TGP considerably decreased HFD-induced body weight gain and serum lipid concentration but improved renal function. In addition, TGP inhibited renal fibrosis and inflammation and reduced MyD88 and p-JNK levels, whereas increased IκB levels. Moreover, silencing MyD88 decreased p-JNK levels while increasing IκB levels which may be the mechanism of TGP treatment. Our findings demonstrated that TGP treatment can ameliorate HFD-induced kidney injury via regulating JNK and IκB signals mediated by MyD88.