ABSTRACT
Objective:To investigate the effect of N-acetylcysteine (NAC) on endoplasmic reticulum stress (ERS) and oxidative stress(OS) induced by tunicamycin (Tm) and its mechanism.Methods:Mouse derived brain microvascular endothelial cells cultured in vitro were divided into control group (normal cell culture), TM group (cells were intervened with 5 μg/mL Tm for 24 h), NAC + TM group (cells were pretreated with 1 mmol/L NAC for 1 h, then were intervened with 5 μg/mL Tm for 24 h) and NAC group (cells were intervened with 1 mmol/L NAC for 24 h) according to different intervention methods.CCK-8 and FITC-Annexin V/PI were used to detect the survival rate and apoptosis rate of cells.Western blot was used to detect the expression of GRP78、CHOP、p-eNOS and caspase-12 protein. Laser confocal microcopy was used to detect the expression of ROS, and colorimetry was used to detect the activity of MDA and SOD.Results:There were significant differences in apoptosis rate and survival rate among the four groups ( F=62.57, 35.00, both P<0.05). The apoptosis rate of TM group ((25.49±1.55)%) was higher than that of Control group ((13.76±1.48)%)( P<0.01), while the apoptosis rate of NAC+ TM group ((17.65±1.00)%) was lower than that of TM group ( P<0.01). The survival rate of TM group ((66.33±5.69)%) was lower than that of Control group ((100.00±2.12)%)( P<0.01), while the survival rate of NAC+ TM group ((85.67±4.04)%) was higher than that of TM group ( P<0.01). Western blot showed that there were significant differences in the expression levels of GRP78、CHOP and p-eNOS among the four groups ( F=32.39, 68.66, 13.12, all P<0.01). The expression levels of GRP78 and CHOP protein in TM group were higher than those of Control group (both P<0.05), while the expression level of p-eNOS was lower than that of Control group ( P<0.01). The expression levels of GRP78 and CHOP protein in NAC+ TM group were lower than those of TM group (both P<0.05), while the expression level of p-eNOS was higher than that of TM group ( P<0.01). There was no significant difference in the expression level of caspase-12 protein among the four groups ( F=0.33, P>0.05). Laser confocal showed that there was significant difference in the average fluorescence intensity of ROS among the four groups ( F=77.66, P<0.01). The average fluorescence intensity of ROS in TM group (32.67±1.53) was higher than that in Control group (12.67±2.08) and NAC+ TM group (18.33±1.53) (both P<0.01). Colorimetry showed that there were significant differences in the activity of SOD and the concentration of MDA among the four groups ( F=40.53, 34.99, both P<0.01). The results of colorimetry showed that the activity of SOD in TM group((41.60±1.53)U/mg) was lower than that in Control group((65.39±4.60)U/mg) and NAC+ TM group((58.72±1.64)U/mg)(both P<0.01). The concentration of MDA in TM((2.27±0.11)μmol/mg) group was higher than that in Control group((1.39±0.13)μmol/mg) and NAC+ TM group((1.44±0.11)μmol/mg) (both P<0.01). Conclusion:NAC can reduce Tm-induced apoptosis of cerebral micro-vascular endothelial cells, which may be related to its inhibition of ERS/ OS-related pathways.
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Objective To explore the expression changes of SIRT1 and related inflammatory regula-tors in peripheral blood of patients with major depressive disorder and analyze the correlation between SIRT1, depression and inflammatory regulators. Methods Forty patients with major depressive disorder and forty healthy controls were selected. Hamilton Depression Scale (HAMD-17) was used to assess the degree of de- pression in patients with depressive disorder. Quantitative Real-time PCR( RT-PCR) was used to detect the relative expression levels of SIRT1,Elf-1,NF-κB,IL-1β,GM-CSF mRNA,and enzyme linked immunosorbent assay(ELISA) was used to detect the expression levels of SIRT1,Elf-1,NF-κB,IL-1β,GM-CSF proteins. The correlation between the severity of depression disorder and SIRT1 and the correlation between SIRT1 and Elf-1 and NF-κB were analyzed. Results (1)Compared with the control group,SIRT1 mRNA expression significantly decreased in the case group (P<0. 01),while Elf-1,NF-κB,IL-1β,GM-CSF mRNA expression significantly increased in the case group (P<0. 01). ( 2) The expression of plasma SIRT1 protein((8. 23± 1. 78)ng/ml) in the case group was lower than that in the control group (P<0. 01). The expressions of plas-ma Elf-1 protein((1 921. 67±271. 07)pg/ml),NF-κB protein((2 057. 29±260. 44)pg/ml),IL-1β protein ((186. 60±31. 00) pg/ml) and GM-CSF protein((183. 69±28. 87) pg/ml) were higher than those in the control group((1 512. 92±284. 54)pg/ml,(1537. 18±313. 82) pg/ml,(144. 79±31. 48) pg/ml,(162. 82± 27. 90) pg/ml,respectively,all P<0. 01). (3) SIRT1 mRNA expression level was negatively correlated with the severity of major depressive disorder (r=-0. 51, P<0. 01) and was negatively correlated with the mRNA expression levels of Elf-1 and NF-κB (r=-0. 66,P<0. 01,r=-0. 64,P<0. 01). Conclusion The expres-sion level of SIRT1 in peripheral blood of patients with major depressive disorder is correlated with the sever-ity of depression. This may be related to the decrease of SIRT1 expression in peripheral blood leukocytes of patients with major depressive disorder,which activates the pathway of NF-κB and Elf-1 and increase expres-sion of GM-CSF and IL-1β.
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Objective To investigate the effect of antidepressants on frontal lobe inflammatory factors in depressed model rats.Methods Thirty SD rats were randomly divided into 5 groups (n=6):contorl group(Cn),depression group (Dn),citalopram group (Dx),venlafaxine group (Dw) and reboxetine group (Dr).The open field test and sugar consumption test were performed to observe the depression behavior and the changes of FGF-1,TNF-α,IL-1 and CRP were measured in frontal lobe of rats by ELISA.Pearson linear correlation analysis was used to evaluate the correlation between behavioral score and inflammatory factors level in rats.Results (1)Compared with the Cn group,the scores of Dn,Dr,Dw,Dx group were decreased on Open field test and sugar consumption test (P＜0.05).Compared with the Dn group,the scores of the Dx,Dw,Dr group were increased in Open field test and sugar consumption test (P＜0.05).(2)Compared with Cn group,the levels of TNF-α,IL-1 and CRP increased in Dn group and FGF-1 level decreased (P＜0.01).Compared with Dn group,the levels of FGF-1 increased in Dx ((86.54±2.56) ng/L),Dw((79.82±4.89) ng/L)and Dr ((68.50 ± 3.61) ng/L) group,however,the levels of TNF-α decreased in Dx ((150.21±5.65) ng/L),Dw ((161.28±8.80) ng/L),Dr ((175.78±9.67) ng/L) group,and the level of IL-1 also decreased in Dx ((30.87±4.48) ng/L),Dw ((36.65±3.33) ng/L),Dr ((40.14±2.81)ng/L) group,and the levels of CRP also decreased in Dx ((374.88 ± 14.15) ng/L),Dw ((394.21 ± 17.03) ng/L),Dr ((414.34± 10.97)ng/L) (P＜0.01).(3)The behavioral score of each group was positively correlated with of FGF-1 and negatively correlated with TNF-a,IL-1 and CRP (P＜0.05).Conclusion Antidepressants can reduce the level of proinflammatory factors and increase the level of anti-inflammatory factors of frontal lobe in depression model rats,suggesting that antidepressants may play an antidepressant effect by regulating the concentration of inflammatory factors.