Transcriptomic analysis to identify genes associated with hypothalamus vulnerability in aging mice with cognitive decline.

The normal aging process is accompanied by cognitive decline, and previous studies have indicated the crucial role of the hypothalamus in regulating both aging and cognition. However, the precise molecular mechanism underlying this relationship remains unclear. Therefore, this present study aimed to identify potential predictors of cognitive decline associated with aging specifically within the hypothalamus. To achieve this, we employed Morris water maze (MWM) testing to assess learning and memory differences between young and aged mice. Additionally, transcriptome sequencing was conducted on the hypothalamus of young and aged mice to identify potential genes. Subsequently, GO and KEGG analyses were performed to investigate the functions of differentially expressed genes (DEGs) and their associated biological pathways. Finally, the results obtained from sequencing analysis were further validated using qRT-PCR. Notably, MWM testing revealed a significant decrease in spatial learning and memory ability among aged mice. According to KEGG analysis, the DEGs primarily encompassed various biochemical signaling pathways related to immune system (e.g., C3; C4b; Ccl2; Ccl7; Cebpb; Clec7a; Col3a1; Cxcl10; Cxcl2; Fosb; Fosl1; Gbp5; H2-Ab1; Hspa1a; Hspa1b; Icam1; Il1b; Itga5; Itgax; Lilrb4a; Plaur; Ptprc; Serpine1; Tnfrsf10b; Tnfsf10), neurodegenerative disease (e.g., Atp2a1; Creb5; Fzd10; Hspa1a; Hspa1b; Il1b; Kcnj10; Nxf3; Slc6a3; Tubb6; Uba1y; Wnt9b), nervous system function (e.g., Chrna4; Chrna6; Creb5; Slc6a3),and aging (e.g., Creb5; Hspa1a; Hspa1b) among others. These identified genes may serve as potential predictors for cognitive function in elderly individuals and will provide a crucial foundation for further exploration into the underlying molecular mechanisms.

Genome-wide analysis of methylation in rat fetal heart under hyperglycemia by methylation-dependent restriction site-associated DNA sequencing.

Diabetes mellitus causes an increased incidence of congenital heart malformations. However, the pathogenesis and potential epigenetic mechanism involved in this process are unclear. In this study, we used MethylRAD sequencing to compare changes in methylation levels in the genomic landscapes in the fetal heart in a rat model of hyperglycemia. Our results showed that methylation of CCGG/CCNGG sites were mostly enriched in intergenic regions, followed by intron, exon, upstream and the 5' and 3' untranslated regions. qRT-PCR results confirmed the MethylRAD sequencing findings, suggesting that abnormal CCGG/CCNGG methylation in the upstream region regulated gene expression. The differential methylation genes (DMGs) based on the CCGG and CCNGG sites in the upstream region were examined by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis. Gene Ontology indicated that the CCGG-based DMGs involved in biological process and function were mainly related to transcription and co-SMAD binding. The CCNGG-based DMGs were mainly related to transcription and cytokine-mediated signaling pathways. Kyoto Encyclopedia of Genes and Genomes analysis indicated that CCGG-based DMGs were mainly involved in the Wnt signaling and TGF-ß signaling pathways. CCNGG-based DMGs were involved in the TNF signaling and apoptosis pathways. These genes may play dominant roles in cardiomyocyte apoptosis and heart disease and require further study. These genes may also serve as potential molecular targets or diagnostic biomarkers for heart malformations under hyperglycemia.

Study on Influencing Factors of Semen Quality in Fertile Men.

Objective: To establish a system for evaluation of semen quality in fertile men by factor analysis (FA). Methods: The FA method was used to analyze five sperm test indicators for fertile men (sperm pH, sperm motility, sperm progressive motility, semen density, and total sperm number) to determine the evaluation standard of semen quality. Pearson analysis was adopted for correlation testing. Results: The comprehensive score formula for semen quality of normal fertile men was as follows: comprehensive score of semen quality = (0.38272 F1 + 0.36359 F2 + 0.20018 F 3)/94.699. Across the whole fertile population, semen quality was found to be correlated with abstinence period, age of first spermatorrhea, and frequency of intercourse. Smoking, drinking, and place of residence were correlated with semen quality in the high semen quality population. In the population with medium semen quality, only the abstinence period was associated with semen quality. Conclusion: It is feasible to evaluate the semen quality of fertile men using the FA method. The comprehensive indicators of semen volume, sperm motility, and semen pH can be used as evaluative measures. Across the whole fertile population, the abstinence period and age of first spermatorrhea were correlated with semen quality. In the high semen quality population, smoking and drinking were negatively correlated with semen quality, and participants living in rural areas had better semen quality.

A Factor Analysis Model for Rapid Evaluation of the Semen Quality of Fertile Men in China.

Objective: The objective of this study is to reduce the dimension of several indicators with a strong correlation when conducting semen quality analysis in a small number of comprehensive variables that could retain most of the information in the original variables. Methods: A total of 1132 subjects were recruited from the Maternal and Child Health Institutions of seven provinces in mainland China. They completed the questionnaire and provided semen samples. Visualization of the correlation between variables was realized by using a function chart and correlation in the PerformanceAnalytics package of the R programming language (version 3.6.3 [2020-02-29]). Factor analysis was conducted using the principal function in the psych package of R. Principal component analysis, combined with varimax rotation, was used in the operation of the model, and two common factors were selected and measured to provide values for the common factor. The score coefficient was estimated using the regression method. Results: The contribution rates of the two common factors to variable X were 43.7% and 33.98%, respectively. When the two common factors were selected, approximately 78% of the information of the original variables could be explained. The correlation coefficients between the first common factor (the quantitative factor) and sperm density, total sperm count, and semen volume were 0.824, 0.984, and 0.544, respectively. The correlation coefficients between the second common factor (the quality factor) and sperm motility and the percentage of forward-moving (progressive spermatozoa) sperm were 0.978 and 0.976, respectively. Conclusion: The correlation between the original variables of a semen quality analysis was strong and suitable for dimensionality reduction by factor analysis. Factor analysis and dimensionality reduction provide a fast and accurate assessment of semen quality. Patients with low fertility or infertility can be identified and provided with corresponding treatments.

Role of EBAF/Nodal/p27 signaling pathway in development of placenta in normal and diabetic rats.

Placentas control the maternal-fetal transport of nutrients and gases. Placental reactions to adverse intrauterine conditions affect fetal development. Such adverse conditions occur in pregnancies complicated by diabetes, leading to alterations in placental anatomy and physiology. In this study, streptozocin (STZ) injection produced sustained hyperglycemia during pregnancy in rats. Hyperglycemic pregnant rats had gained significantly less weight than normal pregnant rats on embryonic day 15.5. We investigated the influence of diabetes on placental anatomy and physiology. Compared with controls, the diabetic group had a markedly thicker junctional zone at embryonic day 15.5. To explore a mechanism for this abnormality, we examined Nodal expression in the junctional zone of control and diabetic groups. We found lower expression of Nodal in the diabetic group. We then investigated the expression of its target gene p27Kip1 (p27), which is related to cell proliferation. In vitro, Nodal overexpression up-regulated p27 protein levels while interfered EBAF up-regulated p27. In vivo, the expression of p27 was lower in diabetic compared with normal rats, and localization was similar between the two groups. In contrast, a higher expression of PCNA was found in diabetic versus normal placenta. Endometrial bleeding associated factor (EBAF), an up-stream molecular regulator of Nodal, was expressed at higher levels in placenta from diabetic versus normal rats. Based on these results, we speculate that the EBAF/Nodal/p27 signaling pathway plays a role in morphological change of diabetic placenta.

Sequencing Analysis of mRNA Profile in Endothelial Cells in Response to ox-LDL.

The pathogenesis of atherosclerosis (AS) and abnormal endothelial cells apoptosis is a multifactorial biological process. Oxidized low density lipoprotein (ox-LDL) is a critical factor in the formation of AS. However, the exact mechanism is still not clear. Therefore, the aim of this study was to investigate some genes and biological pathways in endothelial cells apoptosis in response to ox-LDL. First, our results has validated that ox-LDL is an effective inducer of endothelial cells apoptosis, then, transcriptome sequencing was used to detect differential expression genes. In total, 71 differentially expressed genes (DEGs) were identified, including 32 upregulated genes and 39 downregulated genes. GO analysis showed that DEGs were mainly enriched in cytokine-mediated signaling pathway, gene expression, external side of plasma membrane, steroid binding, and signaling receptor binding. After KEGG analysis, the DEGs mainly focused on the following biochemical signaling pathways, including Signaling molecules and interaction (such as ICOSLG, IL6, ITGAM, TNFRSF13C and VTCN1), Signal transduction (such as IL13RA2, IL6, ITGAM, PDE5A, SGK3 and TNFRSF13C), Immune system (such as FCGR2A, ICOSLG, IL6, ITGAM and TNFRSF13C), and so on. These genes may play a dominant role in HAECs apoptosis and AS genesis. The above prediction and analysis provide an important basis for our follow-up study of the mechanism of these genes, which might be used as molecular targets or diagnostic biomarkers for AS.

Elevated MST1 leads to apoptosis via depletion of YAP1 in cardiomyocytes exposed to high glucose.

BACKGROUND: Gestational diabetes mellitus is a risk factor for congenital heart defects. The article aimed to investigate the expression and roles of MST1, YAP1, Last1/2 and Survivin in modulating HG-induced cardiomyocyte apoptosis and maternal diabetes-induced heart abnormality. METHODS: Diabetes mellitus was induced in rats using streptozotocin. The protein expression and phosphorylation analysis in fetal heart tissue was assessed by western blot and immunohistochemical staining. Hoechst 33342 staining assay was performed to explore H9C2 apoptosis. The gene and protein expression in H9C2 cells was assessed by quantitative PCR and western blot. Knockdown of gene expression was assessed by RNA interference. RESULTS: Our results revealed that increased MST1 protein levels in the heart tissues of the offspring of diabetic rats in vivo and in H9C2 cardiomyocytes under HG treatment in vitro, respectively. Knockdown and overexpression experiments showed that MST1 played a key role in mediating HG-induced apoptosis in cardiomyocytes. Downregulation of YAP1 was associated with HG-induced, MST1-mediated cardiomyocyte apoptosis. Further study showed that MST1 downregulated the protein level of YAP1 through mediation of YAP1 phosphorylation on Ser127 and Ser397; this process also required LATS1/2 participation. MST1 overexpression increased the phosphorylation levels of LATS1/2, which were also shown to be increased in the heart tissues of diabetic offspring. We also found that YAP1 mediated the expression of Survivin during HG-induced apoptosis, and the Survivin-inhibitor YM155 partially inhibited the role of YAP1 in suppressing apoptosis induced by HG in cardiomyocytes. CONCLUSION: These findings reveal a regulatory mechanism of MST1/YAP1/Survivin signaling in modulating cardiomyocyte apoptosis in vitro and maternal diabetes-induced congenital heart defects in vivo.

Downregulation of SOX11 in fetal heart tissue, under hyperglycemic environment, mediates cardiomyocytes apoptosis.

Gestational diabetes mellitus is one of the causes of abnormal embryonic heart development, but the mechanism is still poor. This study investigated the regulatory mechanism and role of SOX11 in congenital heart abnormality in a hyperglycemic environment. Immunohistochemistry, Western blotting, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) showed decreased SOX11 protein and messenger RNA (mRNA) levels in the heart tissue of diabetic offspring compared with the control group. A Sequenom EpiTYPER MassArray showed that methylation sites upstream in SOX11 region 1 were increased in the diabetic group compared with the control group. Luciferase reporter assays and qRT-PCR showed that Dnmt3b overexpression decreased SOX11 promoter activity and its mRNA level, whereas Dnmt3a had little effect on regulating SOX11 expression. Furthermore, we found that Dnmt3L cooperated with Dnmt3b to regulate SOX11 gene expression. Additionally, the function of SOX11 silencing was analyzed by using small interfering RNA-mediated knockdown. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and apoptotic assays showed that SOX11 downregulation inhibited cell viability and induced apoptosis in cardiomyocytes. Overexpression of the SOX11 gene suppressed cardiomyocytes apoptosis after high glucose treatment. We identified a novel epigenetic regulatory mechanism of SOX11 during heart development in a hyperglycemic environment and revealed a distinct role of SOX11 in mediating cardiomyocytes viability and apoptosis.

High-expression of ROCK1 modulates the apoptosis of lens epithelial cells in age-related cataracts by targeting p53 gene.

BACKGROUND: Age-related cataract (ARC) is a serious visual impairment disease, and its pathogenesis is unclear. This article aims to investigate the role of ROCK1 in the apoptosis of lens epithelial cells (LECs) in age-related cataracts. METHODS: We collect anterior capsule samples from normal people, patients with age-related cataracts, young mice and naturally aging cataract mice. The oxidative stress-induced apoptosis model was constructed by cultivating HLE-B3 cells with H2O2. MTT, Hoechst 33342, and TUNEL assay were performed to explore proliferation and apoptosis. HE assay was used to observe cell morphology. The gene and protein expression were assessed by quantitative real-time PCR, western blot, immunofluorescence, and immunohistochemical staining. RESULT: The results from the clinic and mice experiments showed that the numbers of lens epithelial cells from cataract individuals were less than the control individuals. In vitro, the apoptotic cells were increased in lens epithelial cells under H2O2 treatment. The ROCK1 protein level increased in the lens epithelial cells from age-related cataract patients and the old mice, respectively. Meanwhile, the up-regulation of the ROCK1 gene was associated with H2O2-induced HLE-B3 cells apoptosis. MTT and apoptosis assay showed ROCK1 was necessary in mediating H2O2-induced lens epithelial cells apoptosis through ROCK1 over-expression and knockdown experiment, respectively. Further investigation showed that p53 protein levels had been increased during ROCK1-mediated apoptosis in response to H2O2. Besides, ROCK1 phosphorylated p53 at ser15 to up-regulate its protein level. CONCLUSIONS: This study established the novel association of ROCK1/p53 signaling with lens epithelial cells apoptosis and age-related cataract genesis.

The association between iodine intake and semen quality among fertile men in China.

BACKGROUND: Iodine intake is essential in the production of thyroid hormone but very few foods are rich in it. Iodine deficiency or excess iodine level may both lead to thyroid disorders, which further affects human fertility function. The objective of this study is to investigate the relationship between iodine intake and seminal parameters among fertile men in China. METHODS: A total of 1098 couples were recruited by trained physicians at different family planning service stations in 2015. Semen and iodine samples were obtained from male respondents. A questionnaire survey inquired about demographic information from couples. The main outcome variables of semen quality were semen volume, semen concentration, semen motility, and sperm count, and time to pregnancy. Urinary iodine concentration (UIC) was used to measure iodine levels for male respondents. Ordinary least squared regressions and logistic regressions were performed to estimate the association between iodine intake level and semen quality parameters. RESULTS: Male respondents with deficient or excess iodine levels had a 5% higher semen volume relative to those with optimal iodine intake (p < 0.1). Suboptimal iodine intake was negatively associated with semen concentration and semen counts (p < 0.01). Longer time of pregnancy was observed in iodine deficiency and excess group than those in the optimal group (p < 0.01). CONCLUSION: In general, iodine deficiency and excess were both associated with decreasing semen quality parameters in male respondents.

NPY Receptor 2 Mediates NPY Antidepressant Effect in the mPFC of LPS Rat by Suppressing NLRP3 Signaling Pathway.

Accumulated evidences show that neuroinflammation play a pivotal role in the pathogenesis of depression. Neuropeptide Y (NPY) and its receptors have been demonstrated to have anti-inflammative as well as antidepressant effects. In the present study, the ability of NPY to modulate depressive-like behaviors induced by lipopolysaccharides (LPS) in rats and the receptors and signaling mechanisms involved were investigated. Continuous injection LPS (i.p) for 4 days led to development of depressive-like behaviors in rats, accompanied with M1-type microglia activation and increased levels of IL-1ß as well as decreased levels of NPY and Y2R expression in the mPFC selectively. Local injection of NPY into the medial prefrontal cortex (mPFC) ameliorated the depression-like behaviors and suppressed the NLRP3 inflammasome signaling pathway. Y2R agonist PYY (3-36) mimicked and Y2R antagonist BIIE0246 abolished the NPY effects in the mPFC. All these results suggest that NPY and Y2R in the mPFC are involved in the pathophysiology of depression and NPY plays an antidepressant role in the mPFC mainly via Y2R, which suppresses the NLRP3 signaling pathway, in LPS-induced depression model rats.

GSK3ß and MCL-1 mediate cardiomyocyte apoptosis in response to high glucose.

Gestational diabetes mellitus is a risk factor for congenital heart defects. Our previous results indicated that a decrease in myocardial cells and an increase in apoptotic cells leads to heart defects under hyperglycemia, but much work remains to elucidate this important mechanism of myocardial cell apoptosis induced by high glucose (HG). In this study, we found that a decrease in GSK3ß phosphorylation on Ser9 occurred concomitantly with HG-induced cardiomyocyte apoptosis and in the heart tissues of the offspring of diabetic rats in vitro and in vivo. Decreases in GSK3ß (Ser9) phosphorylation in response to HG were remarkably restored after treatment with SC79, an activator of the Akt signaling pathway. SB216763, an effective inhibitor of the GSK3ß signaling pathway, suppressed HG-induced apoptosis in cardiomyocytes. Further studies showed a decrease in the expression of the anti-apoptotic protein MCL-1 was associated with GSK3ß-mediated apoptosis. MCL-1 overexpression partly inhibits HG-induced apoptosis in cardiomyocytes. Herein, this study revealed the roles of GSK3ß and MCL-1 in modulating HG-induced cardiomyocyte apoptosis and maternal diabetes-induced abnormalities.

Pentraxin 3, TNF-α, and LDL-C Are Associated With Carotid Artery Stenosis in Patients With Ischemic Stroke.

Background: Carotid artery stenosis (CAS), typically resulting from atherosclerotic progression, is more common than intracranial atherosclerotic disease in patients with cerebrovascular disease. Prior literature has incompletely established the relationship of Pentraxin 3 (PTX3) and CAS complexity and severity. This study aims to more thoroughly evaluate the association of plasma PTX3 levels and the prevalence and severity of CAS in patients with cerebrovascular disease. Methods: Two hundred and six patients with ischemic stroke underwent multiphase computerized tomography angiography (CTA) of the head and neck to assess the presence and severity of carotid artery stenosis. Patients were divided into groups with either no carotid artery stenosis (CAS-free) or carotid artery stenosis (CAS). The CAS group was further divided into groups based on the degree of stenosis and the number of involved vessels. The PTX3 and Tumor Necrosis Factor-alpha (TNF-α) concentration were measured by ELISA. Results: Plasma levels of PTX3, TNF-α, and low-density lipoprotein cholesterol (LDL-C) were increased significantly in the CAS group patients vs. the CAS-free group (p = 0.000, 0.002, 0.002, respectively). Within the CAS group, PTX3, TNF-α, and LDL-C were significantly elevated in stenosis of ≥50% group compared to <50% group (p = 0.001, 0.002, 0.049, respectively). The multivariate logistic binary regression analysis revealed that increased age, elevated levels of PTX3, LDL-C, and TNF-α were all independent risk factors for occurrence of carotid stenosis. PTX3 level correlated with the severity of carotid stenosis. Conclusions: High plasma PTX3 levels, TNF-α, and LDL-C are significantly correlated with the prevalence and severity of carotid artery stenosis. PTX3 may be a more powerful predictor for the severity of carotid artery stenosis.

Inhibition of GALR1 in PFC Alleviates Depressive-Like Behaviors in Postpartum Depression Rat Model by Upregulating CREB-BNDF and 5-HT Levels.

Estrogen (E2) withdrawal is a core pathology mechanism for postpartum depression (PPD). Galanin (GAL), an estrogen-inducible neuropeptide has also been reported to be associated with depression. However, it still remains unclear which GAL receptors (GALRs) are involved in PPD pathologic process. In the present study, we discovered that the expression of GALR1, rather than GALR2/3, was upregulated with a region-specific pattern in the prefrontal cortex (PFC) of E2 withdrawal induced PPD model rats. Meanwhile, c-fos was also upregulated only in PFC in the same animal model. Injection of GALR1-siRNA into the bilateral PFC ameliorated depressive-like behavior of PPD rats, suggesting that the upregulation of GALR1 in PFC is involved in PPD. Moreover, Western Blot and HPLC assays demonstrated that the downregulation of CREB-BDNF signaling and 5-HT levels in the PFC of PPD rats were reversed after GALR1-siRNA injection. These comprehensive results suggest that the knock down of GALR1 in PFC alleviates depressive-like behaviors and reverse downregulation of CREB-BDNF and 5-HT levels in PPD rat model. HIGHLIGHTS  Expression level of GALR1 mRNA was significantly increased in PFC of estrogen withdraw-induced PPD rats. Injecting GALR1-siRNA into PFC alleviated depressive-like behavior and reversed the decrease of 5-HT level and CREB/BDNF signaling in PFC of PPD rats.

Repeated abortion in adulthood induces cognition impairment in aged mice.

Age-related cognitive decline is one of the major aspects that impede successful aging in humans. Repeated abortion in adulthood can accelerate or aggravate cognitive deficiency during aging. Here we used repeated abortion in female mice adulthood and investigated the consequences of this treatment on cognitive performance during aging. We observed a substantial impairment of learning memory in 15 months old. This cognitive dysfunction was supported by Aß elevation in CA region. Repeated abortion mice have uniform estrous cycles and decreased ERα expression in hypothalamus and hippocampus. Furthermore, repeated abortion not only significantly increased the HMGB1 expression in hippocampus but also increased the plasma and hippocampal protein levels of IL-1ß, IL-6, and TNF-α. Finally, we identified that MPP-induced cell apoptosis and increased HMGB1 expression as well as IL-1ß, IL-6, and TNF-α expression as following Aß elevation. Taken together, our results identify possible molecular mechanisms underlying cognitive impairment during aging, and demonstrated the repeated abortion in adulthood on cognitive function in aged mice.

Injection of oxytocin into paraventricular nucleus reverses depressive-like behaviors in the postpartum depression rat model.

Oxytocin (OXT) has been considered as a neuroregulator mediating social behaviors and stress-related disorders. Recent clinical studies suggest that OXT might also act as antidepressant in postpartum depression (PPD) patients, but the mechanism is still unknown. In the present study, we explored the effect of OXT in paraventricular nucleus (PVN) and possible signaling pathway involved in a PPD rat model induced by gestation restraint stress (GRS). PPD rats exhibited depressive-like behaviors with significantly longer immobility time, shorter climbing time, and lower sucrose consumption compared to the control rats. Plasma corticosterone (CORT) level was also higher in PPD rats. While PVN and supraoptic nucleus (SON) are main OXT synthesis regions in the brain, GRS-induced decrease of mRNA and peptide level of OXT was seen only in PVN. The expression of TrkB in PVN was increased in PPD rats. Local injection of OXT (20ng) into PVN reversed GRS-induced depressive-like behaviors and high plasma CORT level in PPD rats. Moreover, injection of OXT also reversed GRS-induced increase of TrkB in PVN of PPD rats. All those data suggest that OXT plays an antidepressant role by, at least in part, modulating HPA axis via TrkB in PVN of PPD rats.

Role of GAB1/PI3K/AKT signaling high glucose-induced cardiomyocyte apoptosis.

Gestational diabetes mellitus (GDM) is a risk factor for abnormal heart development. Previous work showed that a decrease of myocardial cells and an increase of apoptotic cells leading to heart defects under hyperglycemia, and many genes and protein have been found to play important roles in cardiomyocyte apoptosis. However, there are still many blind nodes in HG-induced cardiac apoptosis. Our study showed that down-regulation of GAB1 occurred concurrently with HG-induced cardiomyocytes apoptosis and in the heart tissues of offspring of diabetic rats in vitro and in vivo. MTT and apoptosis assay showed GAB1 played a key role in mediating HG-induced apoptosis of cardiomyocytes. Down-regulation of XIAP and increased activities of Caspase3/7 was associated with GAB1-mediated cardiomyocyte apoptosis in response to HG treatment. Further study showed that the phosphorylation levels of AKT (Ser473) decreased after HG treatment. Over-expression of GAB1 resisted the reduction in AKT phosphorylation in response to HG. LY294002, which is an effective inhibitor of the PI3K/AKT signaling pathway, partly inhibited GAB1 to suppress apoptosis induced by HG in cardiomyocytes, and partly suppressed GAB1 to resist the decrease of XIAP in response to HG, indicating AKT signaling, XIAP, and Caspase3/7 participated in GAB1-mediated cardiomyocyte apoptosis in response to HG. Generally, we demonstrate a novel role of GAB1 and its down-stream signaling PI3K/AKT for modulating cardiomyocyte apoptosis in response to high glucose in vitro and vivo.

Involvement of retinoblastoma-associated protein 48 during photodynamic therapy of cervical cancer cells.

5-Aminolevulinic acid-mediated photodynamic therapy (ALA­PDT) is an effective treatment option for cervical intraepithelial neoplasia, the precancerous lesion of cervical cancer, and early cervical cancer, particularly for young or nulliparous women who want to remain fertile. A previous report described the involvement of histone deacetylases (HDAC) during ALA­PDT mediated apoptosis in the cerebral cortex of a mouse model. Retinoblastoma­associated protein 48 (RbAp48), a highly abundant component of HDACs, is a critical mediator that controls the transforming activity of human papillomavirus 16 in cervical cancer cells. The aim of the present study was to investigate the involvement of RbAp48 in ALA­PDT­induced cell death in cervical cancer cells. RbAp48 was significantly upregulated in cervical cancer cell lines treated with ALA­PDT, including SiHa and HeLa cells. To establish the relevance of RbAp48 and the efficacy of ALA­PDT in cervical cancer cells, the effect of ALA­PDT was investigated in SiHa or HeLa cells following the depletion of RbAp48 by small interfering RNA (siRNA). Reduction of RbAp48 led to the reduced suppression of proliferation and apoptosis induced by ALA­PDT in cervical cancer cells, which was associated with a reduction in tumor suppressor protein 53 (p53), retinoblastoma (Rb), apoptosis­related enzyme caspase­3, and increased levels of the oncogenic genes, human papillomavirus E6 and E7. These results provide evidence that RbAp48 is an important contributor to the efficacy of ALA­PDT in cervical cancer cells. RbAp48 may be a therapeutic target that may help to improve the treatment of cervical cancer.

ROCK1/p53/NOXA signaling mediates cardiomyocyte apoptosis in response to high glucose in vitro and vivo.

Gestational diabetes mellitus is a risk factor for congenital heart defects; however, the molecular basis of the congenital heart anomalies remains obscure. Previous reports showed a positive correlation between abnormal cardiomyocyte apoptosis and ventricular wall thinness, one type of congenital heart anomaly. This work explored the expression pattern and molecular mechanism of the Rho-associated coiled-coil containing protein kinase 1 (ROCK1) gene in cardiomyocyte apoptosis and genesis of ventricular wall thinness. In this report, we found a marked increase in the number of apoptotic cardiomyocytes in response to high glucose (HG) treatment. Moreover, up-regulation of ROCK1 expression observed in diabetic offspring compared with controls was potentially associated with cardiomyocyte apoptosis and the ventricular wall thinness. Further investigation showed that p53 and NOXA protein levels increased during ROCK1-meidated apoptosis in response to HG. In response to HG, whereby ROCK1 phosphorylated p53 at Ser15 to up-regulate its protein level. Furthermore, we found that p53 mediated the expression of NOXA during HG-induced apoptosis, and histone acetyltransferase p300 participated in this process. These findings reveal a novel regulatory mechanism of ROCK1/p53/NOXA signaling in modulating cardiomyocyte apoptosis in vitro and maternal diabetes-induced congenital heart defects in vivo.

Down-regulation of GJA3 is associated with lens epithelial cell apoptosis and age-related cataract.

Lens epithelial cell apoptosis is regarded as the common molecular basis of the initiation and subsequent progression of cataract. Recent studies have shown that Oxidative radicals derived from H2O2 causes lens epithelial cell apoptosis, While much work still needs to be done to elucidate this important mechanism of lens epithelial cell apoptosis induced by H2O2. The present study investigated the effect of human lens epithelial cell (SRA01/04) apoptosis induced by H2O2 and the possible molecular mechanism involved. Our data in this report has validated that H2O2 is an effective inducer of lens epithelial cells apoptosis, with the concentrations of H2O2 (100 µM). Moreover, we revealed that the down regulation of the GJA3 gene was associated with H2O2-induced lens epithelial cells apoptosis. Over-expression of GJA3 gene restrained the lens epithelial cells apoptosis induced by H2O2. Furthermore, GJA3 V44 M mutation partly inhibited the capacity of GJA3 to suppress apoptosis induced by H2O2 in SRA01/04 cells, eliciting the critical role of GJA3 in H2O2-induced lens epithelial cells apoptosis. The in vivo results indicated that down-regulation of GJA3 in lens epithelial cells was associated with age-related cataract genesis. Data from this study established the association of GJA3 down regulation with lens epithelial cells apoptosis and age-related cataract genesis.