Antiangiogenic activity of terpenoids from Euphorbia neriifolia Linn.

Angiogenesis is known to serve an important role in embryonic development, wound healing, tissue regeneration, and growth. Two new abietane-type diterpenoids (3, 5), a new lanosterol triterpenoid (8) and seven known compounds haven been isolated from the Euphorbia neriifolia Linn. The structures of all compounds were elucidated by spectroscopic analysis and comparing their NMR data with reported data. Furthermore, we found that compounds 6 and 9 had the antiangiogenic effects in vitro. They could inhibit HUVEC migration and microvessel sprouting in rat aortic rings. Moreover, compound 6 inhibited VEGFR and phosphorylation of Akt, but compound 9 only shown inhibitory effect on phosphorylation of Akt. Taken together, these results suggest that inhibition of VEGF signaling and downstream pathways may be responsible for the antiangiogenic activity of compounds 6 and 9.

[Effects of miRNA-21 on paclitaxel-resistance in human breast cancer cells].

OBJECTIVE: To investigate the effects of miR-21 on paclitaxel-resistance in human breast cancer MCF-7/PR and SKBR-3/PR cells. METHODS: Paclitaxel-resistant human breast cancer cell lines MCF-7/PR and SKBR-3/PR were established by stepwise selection in increasing concentration of paclitaxel. Cellular morphology, mRNA and protein level of MDR1, BCRP and MRP1 in MCF-7/PR and SKBR-3/PR cells were determined. The expression of Bax, Bcl-2 and miR-21 in parental and paclitaxel-resistant cells was detected by RT-PCR and Western blotting. The synthetic miR-21 inhibitor or miR-21 mimic were transfected into MCF-7/PR, SKBR-3/PR and MCF-7, SKBR-3 cells with Lipofectamine 2000. The miR-21 levels were determined by RT-PCR, and P-gp, Bcl-2 and Bax protein levels were examined by Western blotting. MTT assay was used to measure the cell viability, and flow cytometry was performed to analyze the cell cycle and apoptosis. RESULTS: The levels of MDR1, BCRP, MRP1, Bcl-2/Bax and miR-21 in MCF-7/PR and SKBR-3/PR cells were significantly higher than those in MCF-7 and SKBR-3 cells. The protein levels of P-gp, Bcl-2 were up-regulated, and Bax was down-regulated compared with parental cells. MiR-21 was significantly down-regulated after miR-21 inhibitor was transfected; and the levels of MDR1, BCRP, MRP1 and Bcl-2/Bax (P <0.05) were also down-regulated. MiR-21 inhibitors significantly suppressed G0/G1 transition of the cell cycle, and induced cell apoptosis in MCF-7/PR and SKBR-3/PR cells. MTT results showed that miR-21 inhibitors induced sensitivity of MCF-7/PR and SKBR-3/PR cells to paclitaxel. And miR-21 mimic can increase the expression of MDR1, Bcl-2/Bax and change cell morphology from parental cells to resistant cells. RESULTS: The established MCF-7/PR and SKBR-3/PR breast cancer cells show typical multidrug resistance characteristics, which can be used as the model for drug resistance study. Down-regulated miR-21 expression in MCF-7/PR and SKBR-3/PR breast cancer cells can enhance cell sensitivity to paclitaxel.

Centranthera grandiflore alleviates alcohol-induced oxidative stress and cell apoptosis.

Alcohol liver disease (ALD) has become a global threat to human health. It is associated with a wide range of liver diseases including alcohol fatty liver, steatosis, fibrosis and cirrhosis, and finally leads to liver cancer and even death. Centranthera grandiflora is a traditional Chinese medicinal herb commonly used to treat ALD, but no research about its mechanism is available. This study evaluated the hepatoprotective effect and mechanism of C. grandiflora against alcohol-induced liver injury in mice. We found that the ethanol extracts of C. grandiflora (CgW) alleviated the alcohol-induced liver injury, enhanced the levels of antioxidant enzymes, and reduced the amount of lipid peroxides. CgW also affected cell apoptosis by inhibiting the activity of Bax, cleaved-caspase 3 and cleaved-caspase 9, and increasing the activity of Bcl-2. In conclusion, the results showed that CgW can effectively improve ALD through alleviating oxidative stress and inhibiting cell apoptosis for the first time. This study suggested that C. grandiflora is a promising herbal medicine for ALD treatment.

Mimicking Metabolic Disturbance in Establishing Animal Models of Heart Failure With Preserved Ejection Fraction.

Heart failure (HF), the terminal state of different heart diseases, imposed a significant health care burden worldwide. It is the last battlefield in dealing with cardiovascular diseases. HF with preserved ejection fraction (HFpEF) is a type of HF in which the symptoms and signs of HF are mainly ascribed to diastolic dysfunction of left ventricle, whereas systolic function is normal or near-normal. Compared to HF with reduced ejection fraction (HFrEF), the diagnosis and treatment of HFpEF have made limited progress, partly due to the lack of suitable animal models for translational studies in the past. Given metabolic disturbance and inflammatory burden contribute to HFpEF pathogenesis, recent years have witnessed emerging studies focusing on construction of animal models with HFpEF phenotype by mimicking metabolic disorders. These models prefer to recapitulate the metabolic disorders and endothelial dysfunction, leading to the more detailed understanding of the entity. In this review, we summarize the currently available animal models of HFpEF with metabolic disorders, as well as their advantages and disadvantages as tools for translational studies.

Rictor maintains endothelial integrity under shear stress.

Background: Endothelial injury induced by low shear stress (LSS) is an initiating factor in the pathogenesis of various cardiovascular diseases, including atherosclerosis, hypertension, and thrombotic diseases. Low shear stress activates the mammalian target of rapamycin complex 2 (mTORC2) signaling pathway. Rictor, the main constituent protein of mTORC2, is involved in vascular development. However, the impact of conditional Rictor ablation on endothelial homeostasis, especially on endothelial-specific markers, such as vascular endothelial-cadherin (VE-cadherin) and von Willebrand factor (VWF), under blood flow stimulation is unclear. Objective: We aimed to investigate whether endothelial Rictor is involved in maintaining vascular endothelial integrity and the potential role of Rictor in atheroprone blood flow-mediated endothelial injury. Methods and results: Immunofluorescence staining showed that endothelial Rictor was successfully knocked out in a mouse model. Scanning electron microscopy (EM) detection revealed disruption of the endothelial monolayer in the thoracic aorta of Rictor-deficient mice. Furthermore, scanning electron microscopy and transmission electron microscopy showed that Rictor deletion disrupted endothelial integrity and expanded cell junctions in the left common carotid artery region. In vitro, low shear stress disrupted actin filament polarity and the promoted the translocation of vascular endothelial-cadherin, the key component of adherens junctions (AJs) in human umbilical vein endothelial cells. After Rictor downregulation by small interfering RNA, the translocation of vascular endothelial-cadherin and stress fibers increased. Rictor knockdown inhibited low shear stress-induced von Willebrand factor upregulation, and downregulation of vascular endothelial-cadherin decreased low shear stress-induced von Willebrand factor expression. These results suggest that vascular endothelial-cadherin/von Willebrand factor is a possible mechanism mediated by Rictor in the pathological process of low shear stress-induced endothelial injury. Conclusion: Rictor is a key protein that regulates endothelial integrity under vascular physiological homeostasis, and Rictor mediates low shear stress-induced endothelial injury by regulating adherens junctions and von Willebrand factor.

ALDH2 rs671 polymorphism and the risk of heart failure with preserved ejection fraction (HFpEF) in patients with cardiovascular diseases.

Aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism is an established genetic risk of hypertension, diabetes, and coronary heart diseases in Asian population. Previous experimental data showed ALDH2 regulated inflammation, a potential mechanism of heart failure with preserved ejection fraction (HFpEF). However, clinically, the association between ALDH2 polymorphism and incidence of HFpEF remains unknown. In this prospective cross-sectional study, ALDH2 genotyping was performed in 613 consecutive patients enrolled with cardiovascular diseases (CVDs), including hypertension, coronary heart diseases, and/or diabetes mellitus, with normal left ventricular ejection fraction (LVEF). HFpEF was diagnosed according to symptoms and/or signs of dyspnea, fatigue or ankle swelling, N-terminal pro-B-Type natriuretic peptide (NT pro-BNP ≥ 280 pg/mL), LVEF ≥ 50%, and at least one additional criterion: left atrial enlargement (left atrial diameter > 40 mm), diastolic dysfunction (E/E' ≥ 13 or E'/A' < 1) or concurrently with atrial fibrillation. Finally, of 613 patients with CVD, 379 patients (61.8%) were assigned to the wild-type ALDH2*1/*1 group and 234 patients (38.2%) to the mutation-type ALDH2*2 group according to genotyping results. Sixty-nine patients (11.3%) were diagnosed with HFpEF. In ALDH2*2 group, the occurrence of HFpEF was higher (15.4% vs. 8.7%, p = 0.011) than that in ALDH2*1/*1 group. Leukocyte count, the indicator of systemic inflammation, was significantly higher (6.9 ± 2.4 × 109/L vs. 6.5 ± 1.9 × 109/L, p = 0.010) in ALDH2*2 group compared to ALDH2*1/*1 group. In conclusion, ALDH2*2 variant is associated with the risk of HFpEF in patients with CVD. Increased systemic inflammation probably involved in this disease process.

Left atrial diameter and atrial fibrillation, but not elevated NT-proBNP, predict the development of pulmonary hypertension in patients with HFpEF.

BACKGROUND: The determinants of pulmonary hypertension (PH) due to heart failure with preserved ejection fraction (HFpEF) have been poorly investigated in patients with cardiovascular diseases (CVD). METHODS: From July 1 2017 to March 31 2019, a total of 149 consecutive HFpEF patients hospitalized with CVD were enrolled in this prospective cross-sectional study. A systolic pulmonary artery pressure (PASP) > 35 mmHg estimated by echocardiography was defined as PH-HFpEF. Logistic regression was performed to establish predictors of PH in HFpEF patients. RESULTS: Overall, the mean age of participants was 72 ± 11 years, and 74 (49.7%) patients were females. A total of 59 (39.6%) patients were diagnosed with PH-HFpEF by echocardiography. The left atrial diameter (LAD) was related to the ratio of the transmitral flow velocities/mitral annulus tissue velocities in early diastole (E/E') and the left ventricular diameter in systole (LVDs). N-Terminal pro B-type natriuretic peptide (NT-proBNP) was not found to be associated with LAD and impaired diastolic or systolic function of the left ventricle. Multivariable logistic regression showed that atrial fibrillation (AF) increased the risk of PH-HFpEF incidence 3.46-fold with a 95% confidence interval (CI) of 1.44-8.32, P = 0.005. Meanwhile, LAD ≥ 45 mm resulted in a 3.43-fold increased risk, 95% CI: 1.51-7.75, P = 0.003. However, the significance levels of NT-proBNP, age and LVEF were underpowered in the regression model. Two variables, AF and LAD ≥ 45 mm, predicted the PH-HFpEF incidence (C-statistic = 0.773, 95% CI: 0.695-0.852, P < 0.001). CONCLUSIONS: Two parameters associated with electrical and anatomical remodelling of the left atrium were related to the incidence of PH in HFpEF patients with CVD.

Risks of incident heart failure with preserved ejection fraction in Chinese patients hospitalized for cardiovascular diseases.

BACKGROUND: Endogenous aldehyde damages DNA and potentiates an ageing phenotype. The aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism has a prevalence of 30%-50% in Asian populations. In this study, we aimed to analyze risk factors contributing to the development of heart failure with preserved ejection fraction (HFpEF) along with the genetic exposure in Chinese patients hospitalized with cardiovascular diseases (CVD). METHODS: From July 2017 to October 2018, a total of 770 consecutive Chinese patients with normal left ventricular ejection fractions (LVEF) and established CVD (hypertension, coronary heart diseases, or diabetes) were enrolled in this prospective cross-sectional study. HFpEF was defined by the presence of at least one of symptom (dyspnoea and fatigue) or sign (rales and ankle swelling) related to heart failure; N-terminal pro-B-Type natriuretic peptide (NT pro-BNP ≥ 280 pg/mL); LVEF ≥ 50%; and at least one criterion related to elevated ventricular filling pressure or diastolic dysfunction (left atrial diameter > 40 mm, E/E' ≥ 13, E'/A' < 1 or concurrent atrial fibrillation). Logistic regression was performed to yield adjusted odds ratios (ORs) for HFpEF incidence associated with traditional and/or genetic exposures. RESULTS: Finally, among 770 patients with CVD, 92 (11.9%) patients were classified into the HFpEF group according to the diagnostic criteria. The mean age of the participants was 67 ± 12 years, and 278 (36.1%) patients were females. A total of 303 (39.4%) patients were ALDH2*2 variant carriers. In the univariate analysis, eight exposures were found to be associated with HFpEF: atrial fibrillation, ALDH2*2 variants, hypertension, age, anaemia, smoking, alcohol consumption and sex. Multivariable logistic regression showed that 4 'A' variables (atrial fibrillation, ALDH2*2 variants, age and anaemia) were significantly associated with an increased risk of HFpEF. Atrial fibrillation was associated with a 3.8-fold increased HFpEF risk (95% CI: 2.21-6.61, P < 0.001), and the other three exposures associated with increased HFpEF risk were the ALDH2*2 variant (OR = 2.41, 95% CI: 1.49-3.87, P < 0.001), age (OR = 2.14, 95% CI: 1.27-3.60, P = 0.004), and anaemia (OR = 1.79, 95% CI: 1.05-3.03, P = 0.032). These four variables predicted HFpEF incidence in Chinese CVD patients (C-statistic = 0.745, 95% CI: 0.691-0.800, P < 0.001). CONCLUSIONS: 4 A traits (atrial fibrillation, ALDH2*2 variants, age and anaemia) were associated with an increased risk of HFpEF in Chinese CVD patients. Our results provide potential clues to the aetiology, pathophysiology and therapeutic targets of HFpEF.