1.Intergrated analysis of CYP3A5 and ABCB1 gene polymorphisms in renal transplanted recipients treated by Tacrolimus
Jingjing WEI ; Binjian LIU ; Dongmei WANG ; Yan ZHOU ; Hua GUO
Chinese Journal of Organ Transplantation 2015;36(12):736-740
Objective To approach the effect of CYP3A5 6986A>G (rs776746) and ABCB1 3435C>T (rs104564) on plasma tacrolimus(FK506) dose adjusted blood trough concentrations(C0/D) in kidney transplanted patients.Method Totally 125 renal transplanted recipients were recruited,and gene sequencing method were used to detect the genotype of CYP3A5 6986A>G and ABCB1 3435C> T.FK506 trough concentration of 125 patients was detected by enzyme multiplies immunoassay technique (EMIT) after the surgery over 6 month.The relevance between the ratio of trough concentration/dosage and polymorphisms was analyzed.Result In 125 renal transplanted receipients,the allele frequency of CYP3A5 and ABCB1 genetic variants was 74% and 34.8%,respectively.After 6 months,the C0/D ration in the homozygous mutant CYP3A5 * 3/* 3 receiptients was significant higher than that in the homozygous wild type * 1/* 3 and heterozygous mutant * 1/* 1 receiptients (P < 0.05).And there was no significant difference of the Co/D ration between CYP3A5 * 1/* 1 and CYP3A5 * 1/* 3 receipients(P>0.05).The the C0/D ration in homozygous wild type ABCB1 T/T and homozygous mutant T/C receipients was significant higher than that in heterozygous mutant C/C receipients (P<0.05),and there was no significant difference of the C0/D ration between T/C and T/T receipients (P> 0.05).Conclusion The study shows that the the polymorphism of CYP3A5 and ABCB1 genes was significantly correlated with the trough C0/D value of the recipients after renal transplantation who were treated by Tacrolimus for more than half a year.The detection of the SNP of the CYP3A5 and ABCB1 will be useful for FK506 dosage adjustment.
2.Effect of keratinocyte growth factor receptor transgene on sodium channel in alveolar type Ⅱcells with LPS-induced acute hmg injury
Binjian LIU ; Ghaoling QI ; Shuhui ZHENG ; Muxiu ZHOU ; Jianming WANG
Chinese Journal of Emergency Medicine 2009;18(5):457-461
Objective To explore the effects of keratinocyte growth factor receptor (KGFR) transgene on sodium channel in alveolar type Ⅱ cells with LPS-induced acute lung injury, to provide the evidence for gene treatment in acute lung injury. Method Totally 40 male Sprague-Dawtey rats were randomly divided into four groups, including normal control (n=8), injured control (n=10), normal transgene (n=10) and injured transgene (n=12). The models of acute lung injury were produced using LPS, and the successful criteria was the obvious enlargement in the lung tissue. The rats in normal transgene group and injured transgene group were injected with 1 mL of KGFR adenovirus vector through rats' tail vein. At 72 hours later, the rats in injured control group and injured transgene group were injected with LPS in dose of 5 mg/kg (BW). While rats in normal control group and normal transgene group were injected with equivalent saline simultaneously. Another 48 hours later, rats in the four groups were killed. The lung tissue were collected for analysis. The expression of sodium channel in rats' alveolar type Ⅱ cells were detected by immunohistochemistry and immunoeectron microscope. Difference among the experimental groups were estimated by ANOVA analysis (LSD-t-test). There was statistical signifi-cance when P<0.05. Results The levels of sodium channel expression in rats' alveolar type Ⅱ cells were differ-era, with normal control group (47.7±3.33), normal transgene group (46.9±5.21), injured tramgene group (29.19±4.11) and injured control group (5.1±2.3). The level of sodium channel expression in injured trans-gene group was lower than that in normal transgene group (t=9.134, P<0.001) and normal control group (t=10.601,P<0.001), but signifieantly higher than that in injured control group (t=16.466, P<0.001). Conclusions The transgene vector can effectively promote the expression of sodium channel in alveolar type Ⅱ cells in rats with LPS-indueed acute lung injury, and can alleviate sodium and water reteraion in alveolar.
3.Effects of Houttuynize Herba on IκBα/NF-κB signaling pathway by regulating intestinal flora in mice with gastric ulcer
Xinchen SONG ; Haiyan WANG ; Guoli CUI ; Xuemei MA ; Binjian ZHOU ; Qingtian WU
International Journal of Traditional Chinese Medicine 2024;46(12):1593-1600
Objective:To investigate the effects of Houttuynize Herba decoction on aspirin-induced gastric ulcer (GU) in mice; To discuss its mechanism.Methods:A total of 64 SPF male mice were selected, and 48 mice were randomly selected to establish the model by gavage of 20 mg/ml aspirin solution. The remaining 16 rats were treated as normal group by gavage with the same amount of normalsaline once a day for 7 consecutive days. After successful modeling, the remaining mice in the model group were randomly divided into 5 groups, with 8 mice in each group, namely normal saline group (given normal saline), omeprazole group (given omeprazole 0.5 mg/ml), Houttuynize Herba high-, medium- and low-dosage groups (given 1.08 g/ml, 0.54 g/ml, 0.27 g/ml), and the remaining 8 mice in the normal group were given the same amount of normal saline by gavage. The mice were treated by gavage once a day for 7 days. The number of Escherichia coli and Bifidobacterium in mouse feces was counted by bacterial culture method, and the ratio of Bifidobacterium to Escherichia coli (B/E value) was used to judge the imbalance of bacterial flora. The expression of interleukin-6 (IL-6) in serum was detected by magnetic particle chemiluminescence. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of prostaglandin E 2 (PGE 2) in the serum of mice to determine the level of inflammation. Hematoxylin-eosin staining was used to determine the ulcer and healing. ImageJ 1.8.0 was used to calculate the ulcer inhibition rate. The protein expression levels of IκBα, NF-κB-p65 subunit and phosphorylated IκBα (p-IκBα) and p65 (p-NF-κB-p65) subunits in gastric tissue of mice were evaluated by Western blot. Results:Compared with the normal group, the epithelial cells of the gastric mucosa were missing, the glands were irregularly arranged, and the tissue structure was severely damaged in the modeling group; the number of Escherichia coli in the intestine increased ( P<0.01), the number of Bifidobacterium decreased ( P<0.01), and the B/E value was less than 1 ( P<0.01); Serum PGE2 levels were decreased ( P<0.01), IL-6 levels were increased ( P<0.01); The expression of p-IκBα and p-NF-κB-p65 proteins in gastric tissues was elevated ( P<0.05). After 7 days of drug treatment, compared with the saline group, gastric mucosal cells and structures were improved, and weight gained in the Houttuynize Herba groups ( P<0.05); the rate of inhibition of ulcers in mice in the Houttuynize Herba high-dosage group was significantly improved ( P<0.01); the number of Bifidobacteria in the intestinal tract significantly increased ( P<0.01), that of Escherichia coli was diminished ( P<0.05), B/E value was greater than 1 ( P<0.05), IL-6 content in peripheral blood was reduced ( P<0.05), PGE 2 levels significantly increased ( P<0.01); the level of p-IκBα/IκBα and p-NF-κB-p65/NF-κB-p65 in the gastric tissues of mice decreased ( P<0.01). Conclusion:Houttuynize Herba decoction can effectively improve gastric mucosal injury in mice, and its mechanism may be related to regulating intestinal microorganisms, inhibiting the opening of IκBα/NF-κB pathway, and reducing inflammatory response.