1.Up-regulation effect of cyclosporine A on level of kidney inj ury molecule-1 in culture supernatant of human kidney cells and its mechanism
Lianlian SONG ; Jun ZHAO ; Jinyu YU ; Wenlan ZHANG ; Lijuan XUE ; Yaowen FU
Journal of Jilin University(Medicine Edition) 2014;(6):1201-1205
Objective To investigate the mechanism of increasing of the level of kidney injury molecule-1(KIM-1)in culture supernatant of human kidney cells(HKC)induced by cyclosporine A(CsA),and to clarify the relationships between the expression levels of KIM-1 and p38 MAPK pathway and ERK1/2MAPK pathway in HKC. Methods The HKC at logarithmic growth phase were randomly divided into control group, CsA control group, CsA + p38 kinase inhibitor group, p38 kinase inhibitor group, CsA + ERK1/2 inhibitor group and ERK1/2 kinase inhibitor group.The inhibitory rates of proliferation of HKC in various groups were detected by MTT assay, and the expression levels of KIM-1 in HKC supernatant in various groups were detected by ELISA;the survival rates,apopototic rates and necrotic rates of the HKC in various groups were detected by flow cytometry. Results Compared with control group,the expression level of KIM-1 protein in the supernatant of HKC in CsA control group was significantly increased (P<0.05),and the survival rate was significantly decreased (P<0.05), while the apoptotic rate and the necrotic rate were significantly increased (P<0.05 ). Compared with control group,the survival rates, the apoptotic rates and the necrosis rates of cells in p38 kinase inhibitor group and ERK1/2 kinase inhibitor group had no significant differences(P>0.05).Compared with CsA control group,the expression levels of KIM-1 protein in CsA+ p38 kinase inhibitor group and CsA+ ERK1/2 kinase inhibitor group were significantly decreased (P<0.05),and the survival rate was significantly increased (P<0.05),while the apoptotic rate and the necrotic rate were significantly decreased (P<0.05).Conclusion p38 MAPK pathway and ERK1/2MAPK pathway are involved in the process of up-regulation of the KIM-1 level in HKC culture supernatant induced by CsA,and the expression of KIM-1 may become the biochemical marker of clinical monitoring of CsA nephrotoxicity.
2.Effect of metformin on glucolipid metabolic disorders caused by olanzapine and quetiapine in rats
Jiezheng DONG ; Lianlian XU ; Yi LIU ; Dange SONG ; Shuzhen LI ; Wenjing ZHU ; Xiwen HU
Chinese Journal of Pharmacology and Toxicology 2014;(3):362-366
OBJECTIVE Tostudytheeffectofmetforminonglucolipidmetabolicdisorderscaused byolanzapineorquetiapineinrats.METHODS Olanzapine1mg·kg-1·d-1wasgivenigorquetiapine 20 mg·kg -1·d -1 was given ig for 4 d,and the dose increased to 40 mg·kg -1·d -1 from the 5th day.Met-formin 100 mg·kg -1·d -1 was given ig from the 15th day.The treatment lasted 8 weeks.Body mass, fasting blood sugar (FBS)and postprandial 2 hours blood glucose (2hPBG)were measured at base-line,3 d,1 week,2 week,4 week,6 week and 8 week.At the end of the 8th week,serum cholesterol (TC),triglyceride (TG),low density lipoprotein (LDL-C),high density lipoprotein (HDL-C),fruc-tosamine(FA)andinsulin(IRS)weremeasured.RESULTS Therewasnosignificantstatisticaldiffer-ence between normal control group and metformin 1 00 mg·kg -1·d -1 group.At the end of the 6th week, compared with normal control group,the body mass and 2hPBG were significantly increased in olanzap-ine 1 mg·kg -1·d -1 group and quetiapine 40 mg·kg -1·d -1 group (P<0.05),respectively.At the end of the 8th week,body mass,2hPBG,INS,FA,TC,TG,LDL-C were significantly increased (P<0.05), and HDL-C decreased in olanzapine group and quetiapine group(P<0.05),respectively.FBS was increased only in olanzapine group(P<0.05).Compared with olanzapine group or quetiapine group, body mass,FBS,2hPBG,INS,FA,TC,TG,LDL-C were significantly decreased by metformin admin-istration(P<0.05).CONCLUSION Metformincaneffectivelypreventandtreatweightgainand glucolipid metabolic disorder caused by olanzapine or quetiapine.
3.Function and mechanism of TRIM22 targeting eIF4E in the process of NB4 cells differentiation
Yang HAN ; Guanhua SONG ; Jing TIAN ; Qiong LIAO ; Lianlian LI ; Xiaoyu ZHANG ; Hongyan LIU ; Zhiyong ZHANG ; Guosheng JIANG
Journal of International Oncology 2017;44(4):251-256
Objective To investigate the function of tripartite motif protein 22 (TRIM22) and the interaction with eukaryotic translation initiation factor-4E (eIF4E) in the differentiation of NB4 cells, one kind of acute promyelocytic leukemia cells, which elucidates the mechanism of TRIM22 targeting to regulate eIF4E.Methods The model of NB4 cells inducing differentiation was established in vitro.The expression changes of gene and protein of TRIM22 and eIF4E were detected by using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.In addition, the effect on cell function and protein expression level of eIF4E after adopting electroporation technology to depress or over-express TRIM22 was detected by CCK-8 and flow cytometry.Finally, the interaction of TRIM22 and eIF4E was verified by using co-immunoprecipitation (CO-IP).Results The mRNA relative expression level of TRIM22 was gradually increasing from 1.01±0.15 to 30.98±2.79 (F=280.700, P=0.000), and the protein relative expression level was gradually increasing from 0.22±0.03 to 0.51±0.05 (F=51.430, P=0.000) after the all-trans-retinoic acid (ATRA) induction for NB4 cell.However, the mRNA relative expression level of eIF4E was gradually decreasing from 1.01±0.09 to 0.47±0.06 (F=20.520, P=0.000), with the same trend, the protein relative expression level was gradually decreasing from 0.97±0.02 to 0.64±0.09 (F=14.700, P=0.001).The expression level of PE-CD11b in the TRIM22 over-expression group with ATRA detected by flow cytometry [(78.80±2.00)%] was higher than that in the transfection group of empty vetor with ATRA [(58.70±2.70)%] (t=9.535, P=0.000) and the cotransfection group with ATRA [(61.60±3.80)%] (t=8.187, P=0.000).Meanwhile, the protein level of eIF4E changed reversely after over-expressing the gene level of TRIM22 (t=4.985, P=0.007).The CO-IP experiment was used to verify the interaction of TRIM22 and eIF4E.ConclusionTRIM22 is able to promote the cell differentiation during the process of NB4 cells differentiation.Furthermore, eIF4E is a target of TRIM22 for binding with, which plays an important role in depressing the expression of eIF4E.