Association between Longitudinal Change of Sleep Patterns and the Risk of Cardiovascular Diseases.

STUDY OBJECTIVES: To investigate the role of longitudinal change of sleep patterns in the incidence of cardiovascular diseases (CVD). METHODS: Based on UK Biobank, a total of 18,172 participants were enrolled. Five dimensions of healthy sleep including early chronotype, sleep 7-8 hours/day, free of insomnia, no snoring, and no frequent excessive daytime sleepiness were used to generate a healthy sleep score (HSS) ranging from 0 to 5. Corresponding to the HSS of 0-1, 2-3, and 4-5, the poor, intermediate, and healthy sleep pattern were defined. Based on changes of HSS across assessment 1 and 2, we calculated the absolute difference of HSS. For the change of sleep patterns, we categorized five profiles (stable healthy, worsening, stable intermediate, optimizing, and stable poor sleep patterns). The outcomes were incidence of CVD including coronary heart disease (CHD) and stroke. We assessed the adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) by Cox hazard models. RESULTS: Compared with participants with stable poor pattern, those who improved their sleep pattern or maintained the healthy sleep pattern had a 26% and 32% lower risk of CVD, respectively. Stable healthy sleep pattern was associated with a 29% and 44% reduced risk of CHD and stroke. Per unit longitudinal increment of the HSS was related to an 8% lower risk of CVD and CHD. Compared with individuals with constant HSS, those with decreased HSS had a 13% higher risk of developing CVD. CONCLUSION: Optimizing sleep pattern and maintaining the healthy sleep pattern may reduce the risk of CVD.

Frailty, an Independent Risk Factor in Progression Trajectory of Cardiometabolic Multimorbidity: A Prospective Study of UK Biobank.

BACKGROUND: Although frailty was associated with cardiometabolic diseases (CMDs, including coronary heart disease, stroke, and diabetes here), there was no systematic analyses estimating its role in incidence, progression, and prognosis of cardiometabolic multimorbidity (CMM). METHODS: We included 351 205 participants without CMDs at baseline in UK Biobank. Occurrences of first CMD, CMM, and death were recorded. We used multistate models to assess transition-specific role of baseline frailty measured by frailty phenotype and frailty index in CMM progression trajectory from no disease to single CMD, CMM, and death. Association between changes in frailty and outcomes was investigated among 17 264 participants. RESULTS: Among 351 205 participants (44.0% male, mean age 56.55 years), 8 190 (2.3%) had frail phenotype, and 13 615 (3.9%) were moderate/severe frail according to the frailty index. During median follow-up of 13.11 years, 41 558 participants experienced ≥1 CMD, 4 952 had CMM, and 20 670 died. In multistate models, frail phenotype-related hazard ratios were 1.94 and 2.69 for transitions from no CMD to single disease and death, 1.63 and 1.67 for transitions from single CMD to CMM and death, and 1.57 for transitions from CMM to death (all p < .001). Consistent results were observed for frailty index. Improvement of frailty reduced the risk of CMD progression and death. CONCLUSIONS: Frailty is an independent risk factor for all transitions of CMM progression trajectory. Frailty-targeted management is a potential strategy for primary and secondary prevention of CMM beyond chronological age.

The association between sleep characteristics and the risk of all-cause mortality among individuals with cardiometabolic multimorbidity: a prospective study of UK Biobank.

STUDY OBJECTIVES: To investigate the implications of both sleep factors and sleep patterns on the prognosis of cardiometabolic multimorbidity. METHODS: From UK Biobank, individuals with cardiometabolic multimorbidity , defined as the coincidence of at least 2 cardiometabolic diseases (hypertension, diabetes mellitus, coronary heart disease, and stroke) were included in this study. Four low-risk sleep factors, including early chronotype, sleep 7-8 h/d, free of insomnia, and no frequent excessive daytime sleepiness, were used to generate a healthy sleep score ranging from 0 to 4. Participants with a score of 0-1, 2, 3-4 were clustered into groups with poor, intermediate, and healthy sleep pattern, respectively. We assessed the adjusted hazard ratios and 95% confidence intervals for all-cause mortality using the Cox proportional hazards model. RESULTS: Among included 35,757 participants, the mean age (standard deviation)) was 61.82 (6.3) years. After full adjustment, early chronotype, sleep 7-8 h/d, no frequent excessive daytime sleepiness, and free of insomnia were independently associated with 8%, 12%, 11%, and 8% lower risk of all-cause mortality among all persons with cardiometabolic multimorbidity. We found the fully adjusted hazard ratio (95% confidence interval) for all-cause mortality was 0.90 (0.88-0.92) for a 1-point increase in the healthy sleep score. Compared with the reference group, participants with the intermediate and healthy sleep pattern had 9% and 23% lower risk of all-cause death, respectively, in the fully adjusted model. CONCLUSIONS: A healthy sleep pattern combining 4 low-risk sleep factors could be regarded as a healthy lifestyle for individuals with cardiometabolic multimorbidity to lower the risk of all-cause mortality. CITATION: He L, Ma T, Cheng X, Bai Y. The association between sleep characteristics and the risk of all-cause mortality among individuals with cardiometabolic multimorbidity: a prospective study of UK Biobank. J Clin Sleep Med. 2023;19(4):651-658.

Use of fish oil and mortality of patients with cardiometabolic multimorbidity: A prospective study of UK biobank.

BACKGROUND AND AIMS: Cardiometabolic multimorbidity (CMM) has risen as a global issue of public health, with an in-creasing prevalence and more severe clinical prognosis. This study aimed to estimate the association between use of fish oil and mortality among patients with CMM. METHODS AND RESULTS: In this prospective study based on UK Biobank, participants with ≥2 of cardiometabolic diseases (CMDs, including coronary heart disease [CHD], diabetes, hypertension, and stroke in this study) at recruitment were included. Use of fish oil was derived from touchscreen questionnaires at baseline. All-cause and cardiovascular mortality were accessed via electronic health-related records. Kaplan-Meier curves and flexible parametric Royston-Parmar proportion-hazard models were fitted to assess the as-sociations of fish-oil use with all-cause, cardiovascular mortality, and related life expectancy alterations. Among 30 068 participants from UK Biobank (67.9% men; mean age 61.75 years), 5357 deaths were reported during 12.03 years of follow-up. For patients with CMM, use of fish oil was associated with a 17% lower risk of all-cause mortality (95% confidence interval [95% CI] 0.78-0.88, P < 0.001), and 19% lower risk of cardiovascular mortality (95% CI 0.72-0.90, P < 0.001) in multivariable-adjusted models. At 45 years old, using fish oil was associated with 1.66 years of life expectancy gained. CONCLUSION: Among patients with CMM, use of fish oil was associated with a significantly reduced risk of all-cause, cardiovascular mortality, and prolonged life expectancy.

The individual and joint associations of depression and sleep duration with cardiometabolic diseases and mortality: A prospective cohort study.

BACKGROUND AND AIMS: Depression and sleep duration were only mutually adjusted in a few studies, and it is unknown whether these two factors are independent or overlapping risk factors for cardiometabolic diseases (CMDs) and mortality. This study aimed to evaluate the individual and joint associations of depression and sleep duration with CMDs and mortality. METHODS: A total of 261,297 participants who were free of CMD at baseline were included. Sleep duration was divided into three groups (short: <7 h/day, referent: ages 39-64 years: 7-9 h/day; ages 65+ years: 7-8 h/day, and long: ages 39-64 years: >9 h/day; ages 65+ years: >8 h/day). The main outcomes were hypertension, stroke, CHD, DM, all-cause mortality, and cardiovascular mortality. RESULTS: Among the 261,297 participants, depression and short or long sleep duration were independently associated with increased risk of CMDs and mortality (hazard ratio [HR], 1.10-1.38) when they were mutually adjusted, except for the association between short sleep duration and stroke (HR, 1.03; 95% confidence interval [CI], 0.97-1.10). We documented significant additive interactions between depression and short sleep duration in relation to all-cause mortality (relative excess risk due to interaction [RERI], 0.19; 95% CI, 0.02-0.37) and CHD (RERI, 0.30; 95% CI, 0.11-0.48). CONCLUSIONS: In this study, depression and short or long sleep duration were independently associated with an increased risk of CMDs and mortality. We also observed significant additive interactions between depression and short sleep duration in relation to all-cause mortality and CHD.

Association between healthy lifestyle and the occurrence of cardiometabolic multimorbidity in hypertensive patients: a prospective cohort study of UK Biobank.

BACKGROUND: Cardiometabolic multimorbidity (CMM) is becoming increasingly common in patients with hypertension, and it is well established that healthy lifestyle plays a key role in the prevention of hypertension. However, the association between combined lifestyle factors and CMM in patients with hypertension is uncertain. METHODS: This prospective analysis included the data (obtained from the UK biobank) of participants with hypertension who did not have coronary heart disease (CHD), stroke, or diabetes. The outcome was the occurrence of CMM, defined as ≥ 1 disease of CHD, stroke, and diabetes that occurred in participants with hypertension. Four lifestyle factors (smoking, alcohol consumption, diet, and physical activity) were assessed using a weighted healthy lifestyle score, and participants were divided into four groups: the very unhealthy, unhealthy, healthy, and very healthy groups. The flexible parameter Royston-Parmar proportional hazard model was used to estimate hazard ratios (HRs) between lifestyles and CMM, as well as the difference in CMM-free life expectancy. RESULTS: During a median follow-up of 12.2 years, 9812 (18.4%) of the 53,397 hypertensive patients occurred CMM. Compared with the very unhealthy group, the very healthy group had a 41% reduction in the risk for CMM in hypertensive patients and a 32-50% reduction in the risk for specific cardiometabolic diseases such as CHD, stroke, and diabetes. For each lifestyle factor, non-smoking had the greatest protective effect against CMM (HR: 0.64, 95% confidence interval (CI) 0.60-0.68). A lifestyle combining multiple healthy factors extended CMM-free life expectancy (e.g., six years longer at age 45 years for participants in the very healthy group). CONCLUSIONS: Combined healthy lifestyle factors were associated with a lower risk for CMM in hypertensive patients. This suggests that combined healthy lifestyle should be supported to decrease disease burden.

Prophylactic exercise-derived circulating exosomal miR-125a-5p promotes endogenous revascularization after hindlimb ischemia by targeting endothelin converting enzyme 1.

Background: Prophylactic exercise improves clinical outcomes in patients experiencing severe ischemic diseases. Previous studies have shown that exercise could alter the amount or content of circulating exosomes. However, little is known about the role of precursory exercise-derived circulating exosomes (Exe-Exo) in ischemic diseases. We therefore aimed to explore the function and mechanism of Exe-Exo in endogenous revascularization and perfusion recovery in peripheral arterial disease. Methods and Results: We first determined that 4 weeks of precursory treadmill exercise improved perfusion recovery on days 7, 14 and 21 after unilateral femoral artery ligation (FAL) but had no effect immediately after ligation. Then, local muscle delivery of Exe-Exo promotes arteriogenesis, angiogenesis and perfusion recovery, which could be abolished by GW4869, a well-recognized pharmacological agent inhibiting exosome release. This suggests that Exe-Exo mediated exercise-induced revascularization. In vitro, Exe-Exo enhanced endothelial cell proliferation, migration and tube formation. In addition, we identified miR-125a-5p as a novel exerkine through exosomal miRNA sequencing and RT-qPCR validation. Inhibition of miR-125a-5p abrogated the beneficial effects of Exe-Exo both in vivo and in vitro. Mechanistically, these exercise-afforded benefits were attributed to the exosomal miR-125a-5p downregulation of ECE1 expression and the subsequent activation of the AKT/eNOS downstream signaling pathway. Specifically, skeletal muscle may be a major tissue source of exercise-induced exosomal miR-125a-5p via fluorescence in situ hybridization. Conclusions: Endogenous circulating exosomal miR-125a-5p promotes exercise-induced revascularization via targeting ECE1 and activating AKT/eNOS downstream signaling pathway. Identify exosomal miR-125a-5p as a novel exerkine, and highlight its potential therapeutic role in the prevention and treatment of peripheral arterial disease.

Associations between consumption of three types of beverages and risk of cardiometabolic multimorbidity in UK Biobank participants: a prospective cohort study.

BACKGROUND: Although the association between beverages and a single cardiometabolic disease has been well studied, their role in disease progression from the single cardiometabolic disease state to cardiometabolic multimorbidity (CMM) state remains unclear. This study examined the associations between three types of beverages: sugar-sweetened beverages (SSBs), artificially sweetened beverages (ASBs), and pure fruit/vegetable juices, and the incidence of CMM in patients with a single cardiometabolic disease. METHODS: Our analysis included 37,994 participants from the UK Biobank who completed at least one dietary questionnaire and were diagnosed with only one cardiometabolic disease at the time of recruitment. Competing risk models were used to examine the association between the three types of beverages and incidence of CMM. We conducted analysis both in patients with any single cardiometabolic disease and in patients with specific cardiometabolic disease. RESULTS: During a median follow-up of 9.1 years (interquartile range [IQR] 9.0-9.8), a total of 6399 participants developed CMM. The consumption of SSBs and ASBs (>1 serving per day) was associated with a higher risk of CMM (SSBs: hazard ratio [HR] 1.19, 95% confidence interval [95% CI] 1.08-1.31; ASBs: HR 1.15, 95% CI 1.04-1.27). Intake of pure fruit/vegetable juices was inversely associated with the incidence of CMM (0-1 serving per day: HR 0.90, 95% CI 0.85-0.94; >1 serving per day: HR 0.90, 95% CI 0.81-0.99). However, the association of the high-level consumption of pure fruit/vegetable juices (>1 serving per day) was not statistically significant after correcting for multiple testing. In the analysis of patients with specific cardiometabolic diseases, positive associations were observed in patients with hypertension for SSBs consumption, while inverse associations persisted in patients with cardiovascular disease (coronary heart disease or stroke) and in hypertensive patients for pure fruit/vegetable juice consumption. CONCLUSIONS: Consuming >1 serving of SSBs and ASBs per day was associated with a higher risk of CMM in patients with a single cardiometabolic disease. In contrast, intake of pure fruit/vegetable juices was inversely associated with the risk of CMM. Our findings highlight the need to limit the use of SSBs and ASBs in patients with a single cardiometabolic disease.

Association between single and multiple cardiometabolic diseases and depression: A cross-sectional study of 391,083 participants from the UK biobank.

Background: Individual cardiometabolic diseases (CMDs) are associated with an increased risk of depression, but it's unclear whether having more than one CMD is associated with accumulative effects on depression. We aimed to assess the associations between CMDs and depression and determine the accumulative extent. Methods: In this cross-sectional study based on UK Biobank, participants with available information on CMDs and depression were enrolled. The history of CMDs was derived from self-reported medical history and electrical health-related records. Depression status was assessed by the aggregation of self-reported history and antidepressant use, depression (Smith), and hospital inpatient diagnoses. Logistic regression models were fitted to assess the association between the number or specific patterns of CMDs and depression and to test the accumulative effect of CMD number, adjusting for confounding factors. Results: 391,083 participants were enrolled in our analyses. After multivariable adjustments, CMDs of different number or patterns were associated with a higher risk of depression compared with the reference group (all P < 0.001). In the full-adjusted model, participants with one [odds ratio (OR) 1.26, 95% confidence interval (CI) 1.23-1.29], two (OR 1.50, 95% CI 1.44-1.56), and three or more (OR 2.13, 95% CI 1.97-2.30) CMD(s) had an increased risk of depression. A significant, accumulative dose-related relationship between the number of CMDs and depression was observed (OR 1.25, 95% CI 1.24-1.27). The dose-dependent accumulative relationship was consistent in stratified analyses and sensitivity analyses. Conclusions: CMDs were associated with a higher risk of depression, and there was an accumulative relationship between CMD number and depression.

Shift Work and the Risk of Cardiometabolic Multimorbidity Among Patients With Hypertension: A Prospective Cohort Study of UK Biobank.

Background Although the association between shift work and individual cardiometabolic diseases has been well studied, its role in the progression to cardiometabolic multimorbidity (CMM) remains unclear. In this study, we investigate the association between shift work and the incidence of CMM in patients with hypertension. Methods and Results This study is a population-based and prospective cohort study on 36 939 UK Biobank participants. We used competing risk models to examine the association between shift work and the risk of CMM, which was defined as coexistence of hypertension and diabetes, coronary heart disease, or stroke in our study. We also investigated the association between the frequency and duration of shift work and CMM risks. In addition, we conducted a cross-classification analysis with the combination of frequency and duration of shift work, chronotype and sleep duration as the exposure metrics. During a median follow-up of 11.6 years, a total of 5935 participants developed CMM. We found that usually/always night shift workers were associated with a 16% higher risk of CMM compared with day workers (hazard ratio [HR], 1.16 [95% CI, 1.02-1.31]). We also found that a higher frequency of night shifts (>10/month) was associated with increased risk of CMM (HR, 1.19 [95% CI, 1.06-1.34]) that was more pronounced for >10/month in combination with a morning chronotype or <7 hours or >8 hours of sleep duration (HR, 1.26 [95% CI, 1.02-1.56]; HR, 1.43 [95% CI, 1.19-1.72], respectively). Conclusions We find that night shift work is associated with higher CMM risk in patients with hypertension.

Adherence to a healthy sleep pattern and incidence of cardiometabolic multimorbidity among hypertensive patients: a prospective study of UK Biobank.

STUDY OBJECTIVES: To investigate whether a healthy sleep pattern would reduce the risk of cardiometabolic multimorbidity (CMM) among hypertensives. METHODS: This is a prospective cohort analysis from the UK Biobank. A total of 69 524 hypertensives without a history of diabetes mellitus, coronary heart disease, or stroke at baseline were enrolled. Five dimensions of healthy sleep at baseline including early chronotype, sleep 7-8 h/d, free of insomnia, no snoring, and no frequent excessive daytime sleepiness were used to generate a healthy sleep score ranging from 0 to 5 (one point was given for each dimension of healthy sleep). A higher score indicated a healthier sleep pattern. We set five groups corresponding to the healthy sleep score of 5, 4, 3, 2, and 0-1, respectively. The primary outcome was the incidence of overall CMM among enrolled hypertensives. We assessed the adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) by Fine-Gray subdistribution hazard models. RESULTS: We found the full-adjusted HR (95% CI) for overall CMM was 0.93 (0.91-0.95) for a 1-point increase in the healthy sleep score. Compared to hypertensives with a healthy sleep score of 0-1, those with a score of 5 had a 27% lower risk of overall CMM, and 37%, 23%, and 20% lower risks of diabetes mellitus, coronary heart disease, and stroke, respectively, after adjusting for sociodemographic characteristic, lifestyle, and clinical factors. CONCLUSIONS: Our results indicated that a healthy sleep pattern was associated with lower risks of CMM outcomes among hypertensives.

Associations of baseline use of fish oil with progression of cardiometabolic multimorbidity and mortality among patients with hypertension: a prospective study of UK Biobank.

PURPOSE: The role of fish oil in the prognosis of hypertensive patients is unknown. This study investigated the associations of fish oil supplementation with the progression of cardiometabolic multimorbidity (CMM) and mortality among patients with hypertension. METHODS: Based on UK Biobank, we enrolled participants with hypertension and free of other cardiometabolic diseases. The exposure was baseline use of fish oil derived from questionnaires at baseline. The primary outcomes were the incidence of CMM and all-cause mortality. Competing risk models and flexible parametric proportion-hazards models were fitted to assess the adjusted hazard ratios (HRs) for the risk of CMM and mortality outcomes, respectively. RESULTS: Among 81,579 participants involved [50.37%, men; mean age, 59.38 years (standard deviation, 7.23 years)], 15,990 CMM events and 6456 all-cause deaths were reported (median follow-up, 12.23 years). In multivariable-adjusted models, baseline use of fish oil was associated with 8% lower risk of CMM [95% confidence interval (95% CI) 0.89-0.96, P < 0.001] and 10% lower risk of all-cause mortality (95% CI 0.85-0.95, P < 0.001). CONCLUSION: In individuals with hypertension, baseline use of fish oil was associated with a reduced risk of CMM and all-cause mortality, and further clinical trials are needed to prove this hypothesis.

Association of healthy sleep pattern with the risk of cardiovascular disease and all-cause mortality among people with diabetes: A prospective cohort study.

AIMS: This study aimed to assess the association of healthy sleep pattern with the risk of cardiovascular disease and all-cause mortality among people with diabetes. METHODS: Our study included 12,770 individuals from the UK Biobank at baseline. Sleep patterns were defined by a combination of five sleep behaviors (chronotype, sleep duration, snoring, insomnia, and excessive daytime sleepiness). The competing risk models were used to estimate the relationship between sleep patterns and CVD (including coronary heart disease [CHD] and stroke) in individuals with diabetes. To examine the association between sleep patterns and all-cause mortality risk, we utilized the flexible parametric Royston-Parmar proportion-hazard models. RESULTS: We recorded 2627 CVD events, which includes 1999 CHD and 903 S events, and 1576 all-cause mortality events. Compared to those with poor sleep pattern, individuals having healthy sleep pattern have a 24% lower CVD risk (p < 0.001), a 26% lower CHD risk (p = 0.001), a 25% lower stroke risk (p = 0.036), and a 21% lower all-cause mortality risk (p = 0.020). CONCLUSIONS: Adherence to healthy sleep pattern has been significantly related to cardiovascular disease and all-cause mortality risk reduction among people with diabetes.

Association of vitamin and/or nutritional supplements with fall among patients with diabetes: A prospective study based on ACCORD and UK Biobank.

Aims: To assess the associations of vitamin and/or nutritional supplements (VNS) with falls among patients with diabetes. Methods: 9,141 and 21,489 middle-aged participants with diabetes from Action to Control Cardiovascular Risk in Diabetes (ACCORD) trial and UK Biobank were included. Use of VNS was collected at baseline, and fall events were recorded using annual questionnaires in ACCORD and electric records in UK Biobank during follow-up. The associations of VNS use with fall risk were analyzed using logistic regression models in ACCORD and Fine-Gray sub-distribution hazard models in UK Biobank. The role of specific supplements was also estimated in UK Biobank, adjusting for confounding factors and multiple comparisons. Results: 45.9% (4,193/9,141, 5.5 median follow-up years) patients in ACCORD and 10.5% (2,251/21,489, 11.9 median follow-up years) in UK Biobank experienced fall and in-patient events during follow-up, respectively. In ACCORD, VNS using was associated with an increased risk of fall (full-adjusted odds ratio [OR]: 1.26, P < 0.05). In UK Biobank, despite no significant association between VNS overall and in-patient fall, vitamin B, calcium, and iron using increased the risk of falls significantly (full-adjusted hazard ratio range: 1.31-1.37, P < 0.05). Conclusions: Use of specific VNS increased the risk of fall among patients with diabetes. The non-indicative use of nutritional supplements for patients with diabetes might be inadvisable.

Endogenous reduction of miR-185 accelerates cardiac function recovery in mice following myocardial infarction via targeting of cathepsin K.

Angiogenesis is critical for re-establishing the blood supply to the surviving myocardium after myocardial infarction (MI) in patients with acute coronary syndrome (ACS). MicroRNAs are recognised as important epigenetic regulators of endothelial function. The aim of this study was to determine the roles of microRNAs in angiogenesis. Eighteen circulating microRNAs including miR-185-5p were differently expressed in plasma from patients with ACS by high-throughput RNA sequencing. The expressional levels of miR-185-5p were dramatically reduced in hearts isolated from mice following MI and cultured human umbilical vein endothelial cells (HUVECs) under hypoxia, as determined by fluorescence in situ hybridisation and quantitative RT-PCR. Evidence from computational prediction and luciferase reporter gene activity indicated that cathepsin K (CatK) mRNA is a target of miR-185-5p. In HUVECs, miR-185-5p mimics inhibited cell proliferations, migrations and tube formations under hypoxia, while miR-185-5p inhibitors performed the opposites. Further, the inhibitory effects of miR-185-5p up-regulation on cellular functions of HUVECs were abolished by CatK gene overexpression, and adenovirus-mediated CatK gene silencing ablated these enhancive effects in HUVECs under hypoxia. In vivo studies indicated that gain-function of miR-185-5p by agomir infusion down-regulated CatK gene expression, impaired angiogenesis and delayed the recovery of cardiac functions in mice following MI. These actions of miR-185-5p agonists were mirrored by in vivo knockdown of CatK in mice with MI. Endogenous reductions of miR-185-5p in endothelial cells induced by hypoxia increase CatK gene expression to promote angiogenesis and to accelerate the recovery of cardiac function in mice following MI.

Induction of microRNA-199 by Nitric Oxide in Endothelial Cells Is Required for Nitrovasodilator Resistance via Targeting of Prostaglandin I2 Synthase.

BACKGROUND: Nitrates are widely used to treat coronary artery disease, but their therapeutic value is compromised by nitrate tolerance, because of the dysfunction of prostaglandin I2 synthase (PTGIS). MicroRNAs repress target gene expression and are recognized as important epigenetic regulators of endothelial function. The aim of this study was to determine whether nitrates induce nitrovasodilator resistance via microRNA-dependent repression of PTGIS gene expression. METHODS: Nitrovasodilator resistance was induced by nitroglycerin (100 mg·kg-1·d-1, 3 days) infusion in Apoe-/- mice. The responses of aortic arteries to nitric oxide donors were assessed in an organ chamber. The expression levels of microRNA-199 (miR-199)a/b were assayed by quantitative reverse transcription polymerase chain reaction or fluorescent in situ hybridization. RESULTS: In cultured human umbilical vein endothelial cells, nitric oxide donors induced miR-199a/b endogenous expression and downregulated PTGIS gene expression, both of which were reversed by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt or silence of serum response factor. Evidence from computational and luciferase reporter gene analyses indicates that the seed sequence of 976 to 982 in the 3'-untranslated region of PTGIS mRNA is a target of miR-199a/b. Gain functions of miR-199a/b resulting from chemical mimics or adenovirus-mediated overexpression increased PTGIS mRNA degradation in HEK293 cells and human umbilical vein endothelial cells. Furthermore, nitroglycerin-decreased PTGIS gene expression was prevented by miR-199a/b antagomirs or was mirrored by the enforced expression of miR-199a/b in human umbilical vein endothelial cells. In Apoe-/- mice, nitroglycerin induced the ectopic expression of miR-199a/b in the carotid arterial endothelium, decreased PTGIS gene expression, and instigated nitrovasodilator resistance, all of which were abrogated by miR-199a/b antagomirs or LNA-anti-miR-199. It is important that the effects of miR-199a/b inhibitions were abolished by adenovirus-mediated PTGIS deficiency. Moreover, the enforced expression of miR-199a/b in vivo repressed PTGIS gene expression and impaired the responses of aortic arteries to nitroglycerin/sodium nitroprusside/acetylcholine/cinaciguat/riociguat, whereas the exogenous expression of the PTGIS gene prevented nitrovasodilator resistance in Apoe-/- mice subjected to nitroglycerin infusion or miR-199a/b overexpression. Finally, indomethacin, iloprost, and SQ29548 improved vasorelaxation in nitroglycerin-infused Apoe-/- mice, whereas U51605 induced nitrovasodilator resistance. In humans, the increased expressions of miR-199a/b were closely associated with nitrate tolerance. CONCLUSIONS: Nitric oxide-induced ectopic expression of miR-199a/b in endothelial cells is required for nitrovasodilator resistance via the repression of PTGIS gene expression. Clinically, miR-199a/b is a novel target for the treatment of nitrate tolerance.

ERß-mediated estradiol enhances epithelial mesenchymal transition of lung adenocarcinoma through increasing transcription of midkine.

Epithelial-mesenchymal transition (EMT) plays a specific role in the migration of tumor cells. Both estrogen and midkine (MK) have been thought to be important factors in promoting the progression of non-small-cell lung cancer (NSCLC) and can enhance EMT. Some evidence indicated the correlation between estradiol (E2) and MK, but the precise mechanism on their interreaction is unknown. Here, we try to clarify whether and how E2 regulates MK expression to promote EMT. We found that E2 increased MK mRNA expression in lung adenocarcinoma cells LTEP-a2 and A549 in a time-dependent manner. E2-induced MK expression was inhibited by the estrogen receptor (ER) antagonist ICI 182,780 and tamoxifen but not by phosphoinositide-3 kinase and MAPK inhibitors, suggesting a genomic mechanism of E2 on the regulation of MK transcription. Moreover, luciferase reporter and chromatin immunoprecipitation assays exhibited that E2 induced ERß recruitment to the estrogen response element in the MK promoter. Small interfering RNA to ERα and ERß revealed that ERß mainly mediated E2-induced MK transcription. Interestingly, E2 enhanced MK expression in accordance with increase of EMT, whereas knockdown of MK could block EMT under E2 stimulation. Importantly, through analyzing lung adenocarcinoma tissues, there was indeed a correlation among levels of E2, MK, and EMT-related protein expression. Taken together, we reported a previously unrecognized mechanism on E2 in the regulation of MK expression and proved that MK plays a pivotal role in progression of E2-regulated EMT.

Stanniocalcin-1 promotes tumor angiogenesis through up-regulation of VEGF in gastric cancer cells.

BACKGROUND: Stanniocalcin-1(STC-1) is up-regulated in several cancers including gastric cancer. Evidences suggest that STC-1 is associated with carcinogenesis and angiogenic process. However, it is unclear on the exact role for STC-1 in inducing angiogenesis and tumorigeneisis. METHOD: BGC/STC cells (high-expression of STC-1) and BGC/shSTC cells (low- expression of STC-1) were constructed to investigate the effect of STC-1 on the xenograft tumor growth and angiogenesis in vitro and in vivo. ELISA assay was used to detect the expression of vascular endothelial growth factor (VEGF) in the supernatants. Neutralizing antibody was used to inhibit VEGF expression in supernatants. The expression of phosphorylated -PKCßII, phosphorylated -ERK1/2 and phosphorylated -P38 in the BGC treated with STC-1protein was detected by western blot. RESULTS: STC-1 could promote angiogenesis in vitro and in vivo, and the angiogenesis was consistent with VEGF expression in vitro. Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro. The process of STC-1-regulated VEGF expression was mediated via PKCßII and ERK1/2. CONCLUSIONS: STC-1 promotes the expression of VEGF depended on the activation of PKCßII and ERK1/2 pathways. VEGF subsequently enhances tumor angiogenesis which in turn promotes the gastric tumor growth.