ABSTRACT
Objective To investigate the effects of Tacrolimus( FK506 )and Puromycin aminonucleoside ( PAN)on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to ex-plore the protective effect of FK506 on podocytes. Methods Mouse glomerular podocytes were cultured in υitro,and the control group,PAN group and FK506 group were established. After 8 h,24 h and 48 h of treatment,the cell mor-phology was observed and the apoptosis rate was detected. The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4. Results The cell body area of the PAN group was significantly smaller than that of the control group,and the cell area of the FK506 group was larger than that of the PAN group. There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h( all P>0. 05). At 24 h and 48 h,the apoptotic rate of podocytes in PAN group[(8. 21 ± 0. 41)%,(16. 32 ± 0. 17)%]were significantly higher than those in the control group[(4. 28 ± 0. 35)%,(6. 27 ± 0. 28)%],and the differences were significant(all P<0. 05). The apoptosis rate of podocytes in FK506 group[(6. 26 ± 0. 24)%,(13. 32 ± 0. 24)%] were significantly lower than those in PAN group,and the differences were significant(all P<0. 05). At 8 h,there was no significant difference in the expression of α -actinin -4 mRNA and protein( all P >0. 05 ). The expression of mRNA(2. 42 ± 0. 21,3. 78 ± 0. 25)and protein(0. 77 ± 0. 04,1. 22 ± 0. 10)in the PAN group was significantly higher than mRNA(1. 50 ± 0. 22,2. 15 ± 0. 15)and protein(0. 44 ± 0. 03,0. 83 ± 0. 07)in the control group at 24 h and 48 h,and the differences were significant(all P<0. 01). The expression of mRNA(1. 65 ± 0. 24,1. 70 ± 0. 32)and protein(0. 52 ± 0. 05,0. 56 ± 0. 07)in FK506 group was significantly lower than that of PAN group,and the differ-ences were significant(all P<0. 05). Conclusions FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4,which provides a basis for the clinical application of FK506 in the treat-ment of glomerular diseases.
ABSTRACT
Objective@#To investigate the effects of Tacrolimus(FK506) and Puromycin aminonucleoside(PAN) on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to explore the protective effect of FK506 on podocytes.@*Methods@#Mouse glomerular podocytes were cultured in vitro, and the control group, PAN group and FK506 group were established.After 8 h, 24 h and 48 h of treatment, the cell morphology was observed and the apoptosis rate was detected.The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4.@*Results@#The cell body area of the PAN group was significantly smaller than that of the control group, and the cell area of the FK506 group was larger than that of the PAN group.There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h (all P>0.05). At 24 h and 48 h, the apoptotic rate of podocytes in PAN group[(8.21±0.41)%, (16.32±0.17)%] were significantly higher than those in the control group[(4.28±0.35)%, (6.27±0.28)%], and the differences were significant (all P<0.05). The apoptosis rate of podocytes in FK506 group[(6.26±0.24)%, (13.32±0.24)%] were significantly lower than those in PAN group, and the differences were significant (all P<0.05). At 8 h, there was no significant difference in the expression of α-actinin-4 mRNA and protein(all P>0.05). The expression of mRNA (2.42±0.21, 3.78±0.25) and protein(0.77±0.04, 1.22±0.10) in the PAN group was significantly higher than mRNA(1.50±0.22, 2.15±0.15) and protein(0.44±0.03, 0.83±0.07) in the control group at 24 h and 48 h, and the differences were significant (all P<0.01). The expression of mRNA (1.65±0.24, 1.70±0.32) and protein (0.52±0.05, 0.56±0.07) in FK506 group was significantly lower than that of PAN group, and the differences were significant (all P<0.05).@*Conclusions@#FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4, which provides a basis for the clinical application of FK506 in the treatment of glomerular diseases.
ABSTRACT
Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis α-actinin 2, Tvα-actinin 2, has been used to diagnose trichomoniasis. This study was undertaken to examine the role of Tvα-actinin 2 as an antigenic molecule to induce immune responses from humans. Western blot analysis using anti-Tvα-actinin 2 antibodies indicated its presence in the secreted proteins of T. vaginalis. ELISA was employed to measure cytokine production by vaginal epithelial cells, prostate cells, mouse dendritic cells (DCs), or T cells stimulated with T. vaginalis or Tvα-actinin 2 protein. Both T. vaginalis and rTvα-actinin 2 induced cytokine production from epithelial cell lines, including IL-10. Moreover, CD4+CD25− regulatory T cells (Treg cells) incubated with rTvα-actinin 2-treated DCs produced high levels of IL-10. These data indicate that Tvα-actinin 2 modulates immune responses via IL-10 production by Treg cells.
Subject(s)
Animals , Humans , Mice , Antibodies , Blotting, Western , Dendritic Cells , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Interleukin-10 , Prostate , T-Lymphocytes , T-Lymphocytes, Regulatory , Trichomonas vaginalis , TrichomonasABSTRACT
Objective To investigate the expression and clinical significance of α-actinin-1 protein (ACTN1) in prostate cancer (PCa) and prostatic hyperplasia (BPH). Methods The clinical data of patients with PCa or BPH treated in our school affiliated hospital were collected between January 2007—October 2014, according to certain criteria. Immunohistochemistry method was used to detect the expression of ACTN1 in 30 samples of PCa and 30 samples of BPH tissues. Western blot assay was used to detect the relative expression of ACTN1 in 18 samples of PCa and 20 samples of BPH tissues in two groups. Results The result of immunohistochemistry showed that the positive expression rates of ACTN1 were 76.7%and 20%in PCa and BPH groups respectively. The difference was statistically significant (P 0.05). There were significant differences in ACTN1 levels between different Gleason score and T staging groups (P<0.05). Conclusion The expression ofα-actinin-1 is significantly higher in PCa tissues than that in BPH tissues. There is the relationship between expression of ACTN1, Gleason scores and T staging.
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Objective To observe the impact of hypoxia/reoxygenation on myocardial cytoskeletal proteins (α-actinin protein,tubulin protein,desmin protein) and to investigate EPO lessening the damage of myocardial cytoskeleton proteins in rats proved by culturing hypoxia/reoxygenation injured myocardial cells in presence of EPO.Methods The rat model of asphyxia-induced cardiac arrest was performed by turning-off the ventilator and clamping the endotracheal tube.After asphyxia for 8 minutes,CPR was carried out.A total of 24 rats were divided into normal group,ischemia/resuscitation (I/R) group and the EPO group (n =8).The model of myocardial dysfunction was determined 2 hours after restoration of spontaneous circulation (ROSC).The rats of EPO group were given EPO 5000 U/kg after ROSC.The rat heart specimens were collected.Actinin,Tubulin and Desmin protein were observed by SABC immunohistochemistry.The cultured cardiomyocytes were taken from neonatal rats and were divided into three groups:the normal group,the hypoxia/reoxygenation (H/R) group (hypoxia 10 h/reoxygenate 4h),the EPO group (hypoxia 10 h/reoxygenate 4 h,plus 10 U/mL EPO).The changes of tubulin and actinin in cultured cardiomyocytes were observe by Immunofluorescence.Results From immunohistochemistry,there were no significant difference in the optical density of actinin,tubulin and desmin among the normal,I/Rand EPO groups.After H/R injury,the structures of the actinin,tubulin protein were destroyed,the network structure of both protein were unclear in cultured myocardial cells.The grades of fluorescence intensity of actinin and tubulin in H/R group were significant lower than those in normal group,but there was no significant difference between H/R group and EPO group.Conclusions The damage of cytoskeleton during ischemia/reperfusion may be time-dependent.EPO has no beneficial effect on the cytoskeleton after I/R injury.