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Objective:To study the effect of Wuzhi capsules on tacrolimus trough concentration in kidney transplant recipients with different CYP3A5 genotypes.Methods:From June 2015 to October 2019, 162 patients who underwent renal transplantation for the first time were retrospectively analyzed. The patients were divided into two groups, combined and uncombined, according to whether combined with Wuzhi capsules. There were 81 cases in the uncombined group (55 males and 26 females), and 81 in the combined group (62 males and 19 females). There was no significant difference between the two groups( P=0.219). The ages of the uncombined group and the combined group were (39.26±11.91) years old and (37.21±10.88) years old ( P=0.103), the weights were (62.39±11.64) kg and (66.18±13.89)kg ( P=0.298), systolic blood pressure were (147.28±20.24) mmHg and (145.00±16.42) mmHg (1 mmHg=0.133 kPa)( P=0.276), diastolic blood pressure were (92.25±13.87) mmHg and (92.20±12.53) mmHg ( P=0.886), alanine aminotransferase were (12.24±8.59) U/L and (17.06±13.11) U/L ( P=0.015), aspartate aminotransferase were (17.76±9.12) U/L and (16.57±8.37) U/L ( P=0.463), fasting blood glucose were (8.70±3.48) mmol/L and (7.18±2.74)mmol/L ( P=0.006), hemoglobin were (98.96±17.53) g/L and (101.05±18.67) g/L ( P=0.789), creatinine were (665.22±296.55) μmol/L and (797.32±279.32) μmol/L ( P=0.007), estimated glomerular filtration rate were (11.47±14.11) ml/(min·1.73m 2) and (8.85±3.71) ml/(min·1.73m 2) ( P=0.130)in the kidney transplant recipients before surgery. Among the 162 cases in this study, there were 86 cases (53.09%) of CYP3A5*1*3 genotype, 17 cases (10.49%) of CYP3A5*1*1 genotype, 59 cases (36.42%) of CYP3A5*3*3 genotype, and the minimum allele frequency of CYP3A5*1 was 37.04%. In the uncombined group, CYP3A5*1*3 genotype 39 cases (48.15%), CYP3A5*1*1 genotype 5 cases (6.17%), and CYP3A5*3*3 genotype 37 cases (45.68%). In the combined group, CYP3A5*1*3 genotype 47 cases (58.02%), CYP3A5*1*1 genotype 12 cases (14.81%), and CYP3A5*3*3 genotype 22 cases (27.16%), with statistically significant differences in the two groups ( P=0.024). The patients were treated with a triple immunosuppressive regimen (tacrolimus+ mycophenolate mofetil+ glucocorticoid) based on tacrolimus [initial dose: 0.15-0.30 mg/(kg·d)], combination of Wuzhi capsules in the combination group (11.25 mg, twice a day). The trough concentration of tacrolimus was detected by enzyme-linked immunosorbent assay, compare the difference in the trough concentration of tacrolimus between the two groups. The relationship between the effect of Wuzhi capsules and CYP3A5 gene polymorphism was compared, and compare the changes before and after the application of CYP3A5 genotype combined with Wuzhi Capsules. The influencing factors of tacrolimus trough concentration were analyzed by multiple linear regression. Results:In the combined with Wuzhi capsules, the dose corrected trough concentration (C 0/D) of tacrolimus was higher than that in patients without Wuzhi capsules, and the extent of increase was related to genotype. The C 0/D of tacrolimus in patients with CYP3A5*3*3 genotype in the combination and non-combination groups were (12.15±2.95) (ng·ml -1/0.1mg·kg -1·d -1) and (9.99±2.33) (ng·ml -1/0.1mg·kg -1·d -1) ( P=0.004), CYP3A5*1*3 genotype were (11.11±3.20) (ng·ml -1/0.1mg·kg -1·d -1) and (6.86±1.62) (ng·ml -1/0.1mg·kg -1·d -1) ( P<0.001), and there were significant difference. However, CYP3A5*1*1 genotype were(8.29±2.64) (ng·ml -1/0.1mg·kg -1·d -1) and (6.16±2.87) (ng·ml -1/0.1mg·kg -1·d -1) ( P=0.160), there was no significant difference. The tacrolimus C 0/D of the combined group before and after the Wuzhi capsule were as follows: CYP3A5*3*3 genotype: (7.18±2.33)(ng·ml -1/0.1mg·kg -1·d -1) and (13.33±3.09) (ng·ml -1/0.1mg·kg -1·d -1) ( P<0.001); CYP3A5*1*3 genotype: (5.14±2.14) (ng·ml -1/0.1mg·kg -1·d -1) and (10.61±3.20) (ng·ml -1/0.1mg·kg -1·d -1) ( P<0.001); CYP3A5*1*1 genotype: (5.17±3.75) (ng·ml -1/0.1mg·kg -1·d -1) and (8.31±2.74) (ng·ml -1/0.1mg·kg -1·d -1)( P=0.002), and the differences were statistically significant. The results of multiple linear regression showed that the combination of Wuzhi capsules (β=0.508, P<0.001) and CYP3A5 genotype(CYP3A5*1*3 and CYP3A5*3*3: β=-0.361, P<0.001; CYP3A5*1*1 and CYP3A5*3*3: β=-0.425, P<0.001)could influence the trough concentration. The sex (β=-0.100, P=0.124) and age (β=-0.003, P=0.967) of renal transplant recipients had no statistical significance to tacrolimus C 0/D. Conclusions:In the renal transplant patients, CYP3A5 genotype and combined use of Wuzhi capsules are the main factors affecting tacrolimus C 0/D. In order to achieve the expected trough concentration as soon as possible, the interaction between CYP3A5 genotypes and drug combination should be considered.
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OBJECTIVE：To inve stigate the effects of polymorphism of CYP3A4，CYP3A5 and other genes on the increasing of tacrolimus blood concentration by Wuzhi capsules in renal transplantation patients. METHODS ：Totally 194 patients in Han nationality underwent renal transplantation and rece ived outpatient follow-up after surgery were selected from the First Affiliated Hospital of Anhui Medical University during Aug. 2015-Nov. 2018，and then divided into single drug group （107 cases）and combination group （87 cases）according to using of Wuzhi capsules or not. Single drug group was given Tacrolimus capsules 0.1-0.15 mg/kg，po，bid+Mycophenolate mofetil capsule s 0.5-0.75 g，bid or Mycophenolate sodium enteric-coated tablets 360-540 mg，bid+Prednisone acetate tablets 10 mg，qd；combination group was additionally given two Wuzhi capsules ，bid，on the basis of single drug group. Two groups were treated at least 7 d. The clinical data （such as patients ’sex and age ）were collected ，and enzyme amplification immunoassay was used to detect steady-state valley concentration of tacrolimus ，and calculate valley concentration/daily dose （C/D）value. PCR was adopted to detect patient genotyping of CYP3A5 gene rs 4646457，rs15524 and rs776746 locus，CYP3A4 gene rs 4646437，rs2242480 and rs 35599367 locus，ABCB1 gene rs 1128503 locus，ABCC2 gene rs3740066 locus，NR1I2 gene rs 3814055 locus，POR gene rs 1057868 locus，PPARA gene rs 4253728 locus，IL-3 gene rs 181781 locus， IL-10 gene rs 1800896 locus，CTLA4 gene rs 4553808 locus，CYP2C19 gene rs 4244285 and rs 4986893 locus，respectively. The correlation of each factor and tacrolimus C/D value was analyzed by Kruskal-Wallis test or Spearman rank test. Multivariate linear regression was conducted. RESULTS ：In 194 renal transplanation patients ，only wild type at rs 3599367 locus of CYP3A4 gene and rs4253728 locus of PPARA gene were detected ，and each genotype distribution of other genes was consistent with the Hardy-Weinberg equilibrium （P＞0.05）. Single factor analysis and multiple linear regression analysis showed that the combination of Wuzhi capsules and rs 776746 polymorphism of CYP3A5 gene were related to tacrolimus C/D value（P＜0.05）. There was statistical significance in tacrolimus C/D values among different genotypes of CYP3A5 gene in single drug group and combnation group（P＜0.05）. C/D value of GG and AG genotype in single drug group were significantly lower than combination group （P＜ 0.05），while there was no statistical significance in tacrolimus C/D value of AA genotype between 2 groups （P＞0.05）. CONCLUSIONS：Combination of Wuzhi capsules or not and polymorphism of CYP3A5 gene rs 776746 locus are associated with blood concentration of tacrolimus in renal transplantation patients in Han nationality. Combined use of Wuzhi capsules can increase blood concentration of tacrolimus in GG and AG genotype ，but have no significant effect on AA genotype.
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OBJECTIVE： To establish a method for the concentration determination of apatinib mesylate in plasma of rats， and to investigate the effects of single and multiple administration of Wuzhi capsules on the pharmacokinetic behavior of apatinib mesylate in rats. METHODS： LC-MS/MS method was used to detect the plasma concentration of apatinib mesylate in rats. Using carbamazepine as internal standard， the determination was performed on Waters XBridge BEH C18 column with mobile phase consisted acetonitrile-0.1％ formic acid solution （45 ∶ 55，V/V） at the flow rate of 0.3 mL/min. The column temperature was 40 ℃. The temperature of injector was 15 ℃， and the sample size was 2 μL. ESI was used for positive ion scanning in MRM mode. The ion pairs m/z used for quantitative analysis were 398.1→212.1 （apatinib mesylate） and 237.2→194.2 （internal standard）. The rats were randomly divided into control group Ⅰ， observation group Ⅰ， control group Ⅱ， observation group Ⅱ， with 6 rats in each group. Control group Ⅰ were given single administration of apatinib mesylate intragastrically （50 mg/kg， similarly hereinafter）. Observation group Ⅰ was given Wuzhi capsules intragastrically （450 mg/kg， similarly hereinafter）， and then 10 min later given apatinib mesylate intragastrically. Control group Ⅱ was given normal saline intragastrically， once a day， for consecutive 7 d， and then were given single administration of apatinib mesylate. Observation group Ⅱ was given Wuzhi capsules intragastrically， once a day， for consecutive 7 d， and then 10 min later were given single administration of apatinib mesylate. The blood samples were collected from intraocular canthus vein plexus and determined 0.25， 0.5， 1.0， 2.0， 2.5， 3.0， 4.0， 6.0， 8.0， 12.0， 24.0 h after intragastric administration， respectively. Pharmacokinetic parameters were apatinib mesylate were calculated and compared among those groups by using DAS 2.1 software and t-test. RESULTS： The linear range of apatinib mesylate were 2-2 000 ng/mL. The lower limit of quantitation was 2 ng/mL. RSDs of intra- day and inter-day were all lower than 10％， and the accuracy were 94.93％-104.68％. Matrix effect did not affect the quantitative analysis of the substance to be measured. Compared with control group Ⅰ， cmax， AUC0-24 h and AUC0-∞ of observation group Ⅰ were increased significantly， CLZ was decreased significantly （P＜0.05）. Compared with control group Ⅱ， AUC0-24 h and AUC0-∞ of observation group Ⅱ were increased significantly， and CLZ was decreased significantly （P＜0.05）. Compared with observation group Ⅰ， AUC0-24 h and AUC0-∞ of observation group Ⅱ were decreased significantly （P＜0.05）. CONCLUSIONS： Established LC-MS/MS method is sensitive and specific， and can be used for the concentration determination of apatinib mesylate in plasma of rats. Wuzhi capsules can influence in vivo pharmacokinetic behavior of apatinib mesylate in rats. The effect of multiple administration of Wuzhi capsules is weaker than that of single administration.
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Objective:To explore the protective effect of Wuzhi capsules on vancomycin-induced kidney injury in rats and investi-gate the action mechanism. Methods:Totally 24 male SD rats were randomly divided into the blank control group,the model control group and the model test group with 8 ones in each. The rats in the model control group and the model test group were intravenously in-jected vancomycin via tail veil,200 mg·kg-1per day for 10 consecutive days,while those in the blank control group were injected sa-line at the same volume. Meanwhile,the rats were orally administered Wuzhi solution(0.25 g·kg-1) in the model test group and sa-line at the same volume in the blank control group and the model control group. The levels of 24-h microalbuminuria(MALB),neutro-phil gelatinase associated lipocalin(NGAL) and kidney injury molecule 1 (KIM-1) in urine were determined,and those of cystatin C (Cys C),serum creatinine (Scr) and blood urea nitrogen(BUN) in serum and those of superoxide dismutase(SOD),malondialde-hyde(MDA) and glutathione peroxidase(GSH-Px) in renal tissues were also determined. Results:The levels of the renal index,Cys C,Scr,BUN,MALB,NGAL,KIM-1 and MDA in the model control group and the model test group were significantly higher than those in the blank control group,and those in the model control group were significantly higher than those in the model test group(P<0.05 or P<0.01). The levels of SOD and GSH-Px in the model control group and the model test group were significantly lower than those in the blank control group,and those in the model control group were significantly lower than those in the model test group(P<0.05). Conclusion:Wuzhi capsules can effectively relieve vancomycin-induced renal injury in rats probably by inhibiting the oxida-tive reaction.
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Objective To investigate the effects of Wuzhi capsules on tacrolimus concentration in heart transplant recipients and provide evidence for individualized dose optimization of tacrolimus.Methods Forty heart transplant recipients receiving Wuzhi capsules were enrolled in this study.Tacrolimus trough concentration was compared before and after coadminstration of Wuzhicapsules.Furthermore,polymorphisms of CYP3A4 * 1G and CYP3A5 * 3 were also detected to clarify correlations between genotypes and effects of Wuzhi capsule.Results Dose-normalized concentration of tacrolimus after coadministartion with Wuzhi capsules was 2.02-fold higher than before,the results of which was not associated with CYP3A4 * 1G and CYP3A5 * 3 genotypes.Wuzhi capsule could significantly decrease the total bilimbin (T-BiL),but not other hepatic and renal function.Conclusion Dose-normalized concentration of tacrolimus in heart transplant recipients is remarkably increased by Wuzhi capsule.The elevated trough levels rarely result in hepatic and renal toxicity.Wuzhi capsule is a safe,effective,and stable drug to increase the trough concentration of tacrolimus.
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OBJECTIVE:To investigate the association of synergistic effects of Wuzhi capsules on tacrolimus with CYP3A5*3 (6986A>G,rs776746) gene polymorphisms. METHODS:One hundred and severty patients underwent renal transplantation receiving tacrolimus maintenance therapy after surgery were selected from our hospital during Jan. 1997-Dec. 2015,and then divided into Wuzhi capsules(+)group(74 cases)and Wuzhi capsules(-)group(96 cases)according to the use of Wuzhi capsules. Both groups received tacrolimus+mycophenolate mofetil+prednisone;Wuzhi capsules (+)group was additionally given Wuzhi capsules,one capsule each time,bid,for more than 12 months. Trough concentration of tacrolimus was detected by CMIA 0,1,3,6,12 months after medica-tion,and the blood concentrations(C0/D)were calculated at different time points after correcting daily dose. CYP3A5*3 gene polymor-phisms was detected by PCR-RFLP. The association of C0/D value with gene polymorphism was investigated by analysis of covariance. RESULTS:Among 170 patients,there were 65 cases of CYP3A5 GG genotype,83 cases of AG genotype and 22 cases of AA geno-type;genotype frequencies were 38.2%,48.8% and 12.9%,which was in line with Hardy-Weinberg balance (P>0.05). There was statistical significance in the distribution frequencies of GG,AG+AA genotype between Wuzhi capsules(+)group and Wuzhi capsules (-)group (P0.05). CON-CLUSIONS:Wuzhi capsules can increase C0/D of tacrolimus in CYP3A5*3 AG+AA genotype,but have no significant effect on C0/D of tacrolimus in GG genotype;CYP3A5*3 genotype should be considered when using Wuzhi capsules as synergist of tacrolimus.
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Objective:To investigate the correlation of ABCB1 polymorphisms and the synergistic effect of Wuzhi capsules on ta-crolimus. Methods:The ABCB11236C>T(rs1128503), ABCB12677G>T/A(rs2032582) and ABCB13435C>T(rs1045642) genotypes were determined by restriction fragment length polymorphism ( RFLP) analysis. The whole blood levels of tacrolimus in renal transplant recipients were measured by chemiluminescent microparticle immunoassay. Analysis of covariance ( ANCOVA) was per-formed to determine the difference of tacrolimus C0/D among the various groups. Results: Whether co-administeration of Wuzhi cap-sules and tacrolimus or not, tacrolimus C0/D of ABCB11236C>T, ABCB12677G>T/A and ABCB13435C>T genotype and haplo-type was without significant difference. Conclusion:When combined with Wuzhi capsules, ABCB11236C>T, ABCB12677G>T/A and ABCB13435C>T mutation is not associated with tacrolimus C0/D.
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Objective:To investigate the rationality of off-label use of tacrolimus combined with Wuzhi capsules in a membranous nephropathy patient. Methods:Based on the related literatures, the application rationality of tacrolimus combined with Wuzhi capsules in a membranous nephropathy patient was analyzed. Results:The combination of tacrolimus and Wuzhi capsules against membranous nephropathy was an off -label drug use, however, it showed certain clinical rationality and economic efficiency. Conclusion:Off-label drug use commonly exists in clinics. Clinical pharmacists should provide evidence support for off-label drug use through searching and analyzing clinical evidences and closely monitoring therapeutic outcome.