ABSTRACT
Cognitive dysfunction is one of the common central nervous systems (CNS) complications of diabetes mellitus, which seriously affects the quality of life of patients and results in a huge economic burden. The glymphatic system dysfunction mediated by aquaporin-4 (AQP4) loss or redistribution in perivascular astrocyte endfeet plays a crucial role in diabetes-induced cognitive impairment (DCI). However, the mechanism of AQP4 loss or redistribution in the diabetic states remains unclear. Accumulating evidence suggests that peripheral insulin resistance target tissues and CNS communication affect brain homeostasis and that exosomal miRNAs are key mediators. Glucose and lipid metabolism disorder is an important pathological feature of diabetes mellitus, and skeletal muscle, liver and adipose tissue are the key target insulin resistance organs. In this review, the changes in exosomal miRNAs induced by peripheral metabolism disorders in diabetes mellitus were systematically reviewed. We focused on exosomal miRNAs that could induce low AQP4 expression and redistribution in perivascular astrocyte endfeet, which could provide an interorgan communication pathway to illustrate the pathogenesis of DCI. Furthermore, the mechanisms of exosome secretion from peripheral insulin resistance target tissue and absorption to the CNS were summarized, which will be beneficial for proposing novel and feasible strategies to optimize DCI prevention and/or treatment in diabetic patients.
ABSTRACT
Objective To investigate the relationship between polymorphisms of gene promoter region INS 5′UTR single nu-cleotide and type 2 diabetes and serum IAA-Ab levels.Methods By Sequenom MassArray SNP genotyping detection technology, INS 3 pyomter regime single nucleotide polymorphisms (rs689,rs714641 77 and rs3842738)of 497 patients in Chongqing with type 2 diabetes cases(treatment group)and 500 cases(control group)were genotyped and analyzed.IAA-Ab levels in diabetes patients was detected.Theχ2 test statistic was used to analyze the treatment group and control groups.The genotype frequency distribution of IAA-Ab-positive and negative groups SNP was analyzed by non-conditional logistic regression,adjusted for sex,age impact,cal-culated the odds ratio (OR)and 95 % confidence interval(CI ).The polymorphic loci with type 2 diabetes susceptibility and serum GAD-Ab levels was evaluated.Results The genotype frequency distribution of rs689AA,TT and AT was 58.75%,28.77% and 12.47%,respectively.The control group are 50.40%,35.60% and 14.00% respectively.The difference was statistically significant (χ2 =3.923,P <0.05).Compared with the genotype of AA,TT genotype can decrease risky of diabetes,with OR values 0.35(95%CI :0.18-1.06).There was significant difference of AA,TT,AT genotypes between IAA-Ab negative and IAA-Ab positive pa-tients (P < 0.05 ).Conclusion INS polymorphisms might be related to the risky of type 2 diabetes and serum IAA-Ab level in chinses population.
ABSTRACT
Objective:To study the influence of protein transduction domain (PTD)-oligomerization domain (OD)-HA fusion proteins on apoptosis of bcr/abl-positive cell lines. Methods:bcr/abl-positive cells were treated with PTD-OD-HA protein. The apoptoses of the cells were detected by flow cytometry (FCM), DNA ladder and transmission electron microscopy (TEM), and the levels of apoptosis-related genes bax and bcl-2 were detected by RT-PCR and Western blotting. Results:FCM examination demonstrated that PTD-OD-HA protein induced the apoptosis of bcr/abl-positive cells; DNA ladder showed that the classic DNA ladders appeared in BaF3-P210 and K562 cells after 48 h treatment with PTD-OD-HA proteins; the apoptoses of BaF3-P210 cells were observed by TEM; the levels of bax in mRNA and protein increased in BaF3-P210 and K562 cells, and bcl-2 decreased. Conclusion:PTD-OD-HA proteins specifically induced the apoptosis of bcr/abl positive cells.
ABSTRACT
Objective: The BCR-ABL fusion gene induced by reciprocal translocation of t (9; 22) (q34; q11) plays an important role in the pathogenesis of chronic myeloid leukemia (CML). Using recombinant ade-noviruses carrying the N-terminal oligomerizaton domain (OD) of the BCR/ABL and chimeric ubiquitin ligase β-TrCP, this study was to investigate the effect of the targeted degradation of oncoprotein BCR-ABL by Ubiqui- tin-Proteasome System on the proliferation of leukemia call line K562. Methods: The recombinant adenovirus-es carrying wild-type β-TrCP gene (Ad5β-TrCP-OD-HA), mutational β-TrCP gene (Ad5 A F-TrCP-OD-HA)and green fluorescent protein gene (Ad5GFP)were amplified in 293 calls and co-infected into K562 cells respec- tively. The rates of infection were analyzed by flow cytometry (FCM). Recombinant protein and BCR-ABL ex-pression was detected by Western blot. Cell proliferation was determined by cell counting and methylcellu- cose clonal cell culture. Cell cycle was observed through FCM. Untreated K562 cells were used as blank con-trols. Result: The leukemia K562 cell lines with exogenous recombinant β-TrCP-OD-HA and F-TrCP-OD-HA gene were established. The infection rates in the three groups were over 66.4% and recombinant protein sus-tained to be expressed. Ad5β-TrCP-OD-HA down-regulated the expression of BCR-ABL and inhibited prolifer-ation of K562 cells. FCM showed that the percentage of cells at S phase was decreased to 10.88%±2.42%, while that of cells at G_0/G_1 was increased to 85.6%±5.61%, with a significant difference (P<0.05). No changes were found in the cell cycle in groups of Ad5 △ F-TrCP-OD-HA and Ad5GFP. Conclusion: There is sustained ex-pression of recombinant β-TrCP-OD-HA protein in K562 cells infected by recombinant adenovirus.β-TrCP-OD-HA could inhibit the proliferation and clonogenicity of K562 cells through targeted degradation of oncoprotein BCR-ABL and arresting the progression of call cycle.
ABSTRACT
Objective To investigate the effects of circadian clock gene Period2(Per2)on the proliferation,differentiation and apoptosis of K562 cells and its probable molecular mechanism.Methods The Per2 expression plasmid pcDNA3.1-Per2 and empty control plasmid were respectively transfected into K562 cells with cationic liposome,and the resistant cells stably expressing Per2 gene were obtained by G418 selection.Their morphological changes were observed under light microscope following Wright-Giemsa staining.Trypan blue excluding staining and MTT assay were employed to evaluate cell proliferation.Flow cytometry was performed to analyze cell cycle distribution and cell apoptosis,and electron microscopy was used to detect cell apoptosis.Meanwhile,the expressions of proliferation and apoptosis associated proteins,such as P53,Cyclin B1 and C-Myc,were respectively detected by RT-PCR and Western blot analysis at mRNA and protein level.Results The K562/Per2 cell line stably expressing Per2 gene was screened out.As compared with either the empty plasmid transfected group(K562/empty)or the untreated group(K562/untreated),K562/Per2 cells was smaller in volume and showed no obvious cellular differentiation.Circadian clock gene Per2 could significantly inhibit both growth and proliferation of K562 cells.The percentage of K562 cells in G2/M phase increased [K562/Per2 group(36.1?5.5)%,K562/empty group(12.5?2.9)%,untreated group(9.7?2.3)%,P