The silencing of ApoC3 suppresses oxidative stress and inflammatory responses in placenta cells from mice with preeclampsia via inhibition of the NF-κB signaling pathway.

OBJECTIVE: Preeclampsia is one of the three primary causes of maternal morbidity and mortality worldwide. This study evaluated ApoC3 in placenta cells of mice with preeclampsia to explore its therapeutic role in preeclampsia and assess its function on oxidative stress and inflammatory responses involving the NF-κB signaling pathway. METHODS: A mouse model of preeclampsia was successfully established. APOC3-siRNA with the best silencing effect was screened out. The expression levels of ApoC3, p65, and IkBα were evaluated. The effect of ApoC3 silencing on metabolic activity and apoptosis was measured. The level of high-sensitivity C-reactive protein (hs-CPR), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α), the activity of matrix metalloproteinase (MMP)-2 and MMP-9, and the expression of malondialdehyde (MDA), 8-isoprostane and oxidized low-density lipoprotein (ox-LDL) were determined. RESULTS: ApoC3-siRNA-3 was the most effective siRNA. The mRNA expression of ApoC3 was scarcely observed, while the expression of p65 decreased and the expression of p-IkBα increased in the ApoC3-siRNA group. Compared with those in the model and empty vector groups, the cell apoptosis rate and the activities of invasion-related factors MMP-2 and MMP-9 increased, while the levels of hs-CPR, IL-6, TNF-α, MDA, 8-isoprostane, and ox-LDL decreased in the ApoC3-siRNA group. CONCLUSION: Silencing ApoC3 could suppress the NF-κB signaling pathway, thereby exercising a protective effect on cell injury induced by oxidative stress and reducing inflammatory responses.

Gadd45a knockdown alleviates oxidative stress through suppressing the p38 MAPK signaling pathway in the pathogenesis of preeclampsia.

OBJECTIVE: Preeclampsia (PE) is a hypertensive disease associated with vascular oxidative stress (OS). Besides, cell response to OS triggers growth arrest and DNA damage inducible alpha (Gadd45a). Herein, we investigated the effect of Gadd45a on OS in PE. METHODS: Umbilical cord tissues and peripheral blood samples were collected from PE patients and normal pregnant women. Immunohistochemistry was applied to identify Gadd45a level and extent of p38 phosphorylation. To verify the effect of Gadd45a on CRL1730 human umbilical vein endothelial cell (HUVEC) behavior, HUVECs were treated with hypoxia/reoxygenation (H/R) and Gadd45a-siRNA or P79350. OS products were detected using thiobarbituric acid-reactive-substance assay, immune enzyme assay, enzyme-linked immunosorbent assay (ELISA) and super oxide dismutase (SOD) kit. HUVEC behaviors were evaluated using flow cytometry, wound-healing test, and Transwell assay. The tube formation ability was assessed using in vitro tube formation assay. RESULTS: PE umbilical cord tissues exhibited increased Gadd45a expression, extent of p38 phosphorylation, 8-isoprostane and oxidatively modified low density lipoprotein (Ox-LDL) and decreased SOD levels. HUVECs treated with H/R or agonist + H/R showed similar expression tendencies of related genes, enhanced apoptosis and inhibited migration, invasion, and blood vessel formation. Gadd45a-siRNA conferred alleviation to OS with decreased apoptosis, greater cell migration, invasion and blood vessel, which appeared to be weakened by treatment of Gadd45a siRNA + P79350. CONCLUSION: Over-expression of Gadd45a and activation of the p38 MAPK signaling pathway are associated with OS in PE. Furthermore, Gadd45a knockdown promotes cell migration and invasion, and the tube formation, thus alleviating OS in PE.

Role of quantitative hepatitis B core antibody levels in predicting significant liver inflammation in chronic hepatitis B patients with normal or near-normal alanine aminotransferase levels.

AIM: Chronic hepatitis B (CHB) patients with normal alanine aminotransferase (ALT) levels are not free from significant hepatic lesions. Recently, there has been an improved understanding of the clinical significance of quantitative hepatitis B core antibody levels (qAnti-HBc) during CHB management. In this cross-sectional study, we evaluated the utility of qAnti-HBc in identifying significant liver inflammation in CHB patients. METHODS: A total of 469 patients (training set, n = 363; validation set, n = 106) who underwent liver biopsy (LB) were included. The qAnti-HBc levels were quantified and the relationship between histology and serum markers was systematically analyzed. RESULTS: In the training set, qAnti-HBc levels were found to have significant diagnostic value for moderate to severe liver inflammation (≥G2) in all patients (area under the receiver operating characteristic curve [AUROC] = 0.768; 95% confidence interval [CI], 0.721-0.810; P < 0.001) and in patients with normal or near-normal ALT levels (AUROC = 0.767; 95% CI, 0.697-0.828; P < 0.001). Our novel index (AC index) for the identification of ≥G2 inflammation, which combined the qAnti-HBc and ALT levels, significantly improved diagnostic performance (AUROC = 0.813; 95% CI, 0.768-0.852) compared to the use of ALT alone (AUROC = 0.779; 95% CI, 0.732-0.821) in all patients. In the validation set, the AC index showed an improved AUROC of 0.890 (95% CI, 0.814-0.942) and 0.867 (95% CI, 0.749-0.943) in all patients and patients with normal ALT levels, respectively. CONCLUSIONS: The qAnti-HBc level predicts significant liver inflammation well, even in patients with normal or near-normal ALT levels. Compared with the conventional ALT level, the AC index is a more reliable non-invasive biomarker for significant liver inflammation in CHB patients.

Arterial stiffness and wave reflections in relation to plasma advanced glycation end products in a Chinese population.

BACKGROUND: Accumulation of advanced glycation end products (AGEs) in the human body might engender arterial stiffening. We investigated the relationship of plasma AGE concentration with arterial stiffness and wave reflections in a Chinese population. METHODS: The study subjects were recruited from a newly established residential area in the suburb of Shanghai in 2009. Using the SphygmoCor system, we measured carotid-femoral pulse wave velocity (cfPWV) and central augmentation indices (cAI) and peripheral augmentation indices (pAI). Plasma AGE concentration was measured by the enzyme-linked immunosorbent assay method and logarithmically transformed for statistical analysis. RESULTS: The 1,051 study participants (mean age = 55.1±13.1 years) included 663 (63.1%) women, 390 (37.1%) hypertensive patients, and 90 (8.6%) diabetic or prediabetic subjects. Plasma AGE concentration was higher in men than women (5.62 vs. 5.07 µg/ml; P = 0.02) and with older age (r = 0.13 in both sexes; P ≤ 0.01) and higher serum total/high-density lipoprotein cholesterol ratio (r = 0.20 in men and r = 0.15 in women; P < 0.0001). In multiple regression analyses, plasma AGE concentration was significantly associated with cAI and pAI (1.9% and 4.0% increase per 10-time increase in plasma AGE concentration, respectively; P ≤ 0.02) but not with cfPWV (P = 0.62). However, there was significant (P = 0.001) interaction between plasma AGE concentration and age in relation to cfPWV. Only in subjects aged ≥70 years, cfPWV increased with higher levels of plasma AGE concentration (bottom vs. top quintile distributions = 8.10 vs. 8.90 m/s; P = 0.02). CONCLUSIONS: AGEs accumulate with aging and high cholesterol and are associated with arterial wave reflections and, in an age-dependent manner, with arterial stiffness.

Urinary angiotensinogen excretion and ambulatory blood pressure.

OBJECTIVE: Previous studies have demonstrated that urinary angiotensinogen excretion, a biomarker of the intrarenal renin-angiotensin system activity, is associated with clinic blood pressure (BP). In the present study, we investigated the determinants of urinary angiotensinogen excretion and its associations with ambulatory BP. METHODS: The study participants were suspected hypertensive patients being off antihypertensive medication for at least 2 weeks and referred to our hypertension clinic for 24-h ambulatory BP monitoring. Ambulatory hypertension was defined as a 24-h BP of at least 130  mmHg systolic or 80  mmHg diastolic. We collected a first morning urine sample for the measurement of angiotensinogen by ELISA kits. RESULTS: The 446 participants (mean age 51.7 years) included 218 (48.9%) men, and 275 (61.7%) patients had ambulatory hypertension. In addition to age and sex, 24-h urinary sodium excretion was an independent determinant of urinary angiotensinogen-to-creatinine ratio (P = 0.0008). Urinary angiotensinogen-to-creatinine ratio was 34% (P = 0.04) and 82% (P ≤ 0.0001) higher in tertiles 2 and 3 of 24-h urinary sodium excretion, respectively, than in tertile 1. In multivariate analyses, urinary angiotensinogen-to-creatinine ratio was significantly and positively associated with clinic and ambulatory BP (P ≤ 0.02) and the prevalence of ambulatory hypertension [odds ratio (95% confidence interval) associated with two-time increase, 1.24 (1.09-1.39); P = 0.0007]. CONCLUSION: Urinary angiotensinogen excretion is higher with greater urinary sodium excretion, and is associated with clinic and ambulatory BP.

Blood flow pattern in the middle cerebral artery in relation to indices of arterial stiffness in the systemic circulation.

BACKGROUND: The brain is perfused at high-volume flow throughout systole and diastole. We explored the association of blood flow in the middle cerebral artery (MCA) with the pulsatile components of blood pressure in the systemic circulation and indices of arterial stiffness. METHODS: We enrolled 334 untreated subjects (mean age, 50.9 years; 45.4% women) who had been referred for ambulatory blood pressure monitoring to Ruijin Hospital, Shanghai, China. We measured the MCA pulsatility index (PI) by transcranial Doppler ultrasonography. The indices of arterial stiffness included pulse pressure (brachial (bPP) and central (cPP) measured at the office and 24-h ambulatory (24-h PP)) and carotid-femoral (cf-PWV) and brachial-ankle (ba-PWV) pulse wave velocity. Effect sizes, expressed per 1 s.d., were adjusted for sex, age, heart rate, and mean pressure. RESULTS: Women had faster MCA blood flow than men (68.0 vs. 58.3 cm/s), but lower PI (75.4 vs. 82.3%; P < 0.001). The five arterial stiffness indices were intercorrelated (r ≥ 0.37; P < 0.001). PI increased (P ≤ 0.045) with bPP (+6.78%), cPP (+5.56%), 24-h PP (+7.58%), cf-PWV (+1.59%), and ba-PWV (+3.46%). In explaining PI variance, bPP ranked first (partial r(2) = 0.25), 24-h PP second (0.20) and cPP third (0.14). In models including both cf-PWV and ba-PWV, only the latter was significant (-0.19%; P = 0.84 vs. +3.54%; P < 0.001). In models including both bPP and ba-PWV, only the former contributed to PI variance (+6.98%; P < 0.001 vs. -0.24%; P = 0.78). CONCLUSION: MCA blood flow is closely associated with the pulsatile pressure in the systemic circulation, which depends on arterial stiffness as measured by PWV.

Peripheral and central augmentation indexes in relation to the CYP4F2 polymorphisms in Chinese.

OBJECTIVE: Cytochrome (CYP) 4F2 isoform is a key metabolizing enzyme for the renal 20-hydroxyeicosatetraenoic acid (20-HETE), which, as an endogenous vasoconstrictor, may influence properties of the peripheral muscular arteries and arterioles. We, therefore, investigated the CYP4F2 polymorphisms in relation to arterial wave reflections, as measured by augmentation indexes (AIx) in Chinese. METHODS: We performed arterial measurements by SphygmoCor and genotyped three CYP4F2 polymorphisms (V433M, rs3093089, and rs3093098) by PCR-restriction fragment length polymorphism in 1421 participants enrolled in the JingNing Population study. A replication study for the V433M polymorphism was performed in 924 Chinese recruited from a workplace setting. Urinary 20-HETE concentration was determined by ELISA in a randomly selected subsample of 318 JingNing individuals. RESULTS: In spite of the fact that genetic associations were not significant (P ≥ 0.12) in all JingNing participants, there was significant (Pint ≤ 0.02) interaction of the V433M polymorphism with sex and pulse rate in relation to peripheral and central AIx. M433 allele carriers, compared with V433V homozygotes, had significantly greater peripheral (+5.0%, P = 0.0002) and central AIx (+3.2%, P = 0.001) in 693 men. The corresponding values were +2.7% (P = 0.04) and +1.9% (P = 0.04) in 490 individuals of the top tertile of pulse rate (≥ 76 beats/min), and were +4.0% (P = 0.02) and +3.3% (P = 0.02) in 315 replication participants with a pulse rate at least 76 beats/min. Urinary 20-HETE concentration was significantly higher (P = 0.002) in M433M (2.06 ng/ml) and V433M (1.13 ng/ml) individuals than in V433V homozygotes (0.98 ng/ml). CONCLUSION: The CYP4F2 V433M polymorphism is associated with the size of arterial wave reflections in male Chinese, or individuals with a faster pulse rate.