Salvianolic acid A alleviates H2O2-induced endothelial oxidative injury via miR-204-5p.

Oxidative stress induced endothelial dysfunction plays a particularly important role in promoting the development of cardiovascular diseases (CVDs). Salvianolic acid A (SalA) is a water-soluble component of traditional Chinese medicine Salvia miltiorrhiza Bunge with anti-oxidant potency. This study aims to explore the regulatory effect of SalA on oxidative injury using an in vitro model of H2O2-induced injury in human umbilical vein endothelial cells (HUVECs). In the study, we determined cell viability, the activities of Lactate dehydrogenase (LDH) and Superoxide dismutase (SOD), cell proliferation rate and intracellular reactive oxygen species (ROS). Flow cytometry was used to detect cell apoptosis. Western-blotting was used to evaluate the expression of cell senescence, apoptosis, autophagy and pyroptosis protein factors. The expression level of miRNA was determined by qRT-PCR. Compared with H2O2-induced HUVECs, SalA promoted cell viability and cell proliferation rate; decreased LDH and ROS levels; and increased SOD activity. SalA also significantly attenuated endothelial senescence, inhibited cell apoptosis, reversed the increase of LC3 II/I ratio and NLRP3 accumulation. Furthermore, miR-204-5p was regulated by SalA. Importantly, miR-204-5p inhibitor had similar effect to that of SalA on H2O2-induced HUVECs. Our results indicated that SalA could alleviate H2O2-induced oxidative injury by downregulating miR-204-5p in HUVECs.

Protocatechualdehyde Rescues Oxygen-Glucose Deprivation/Reoxygenation-Induced Endothelial Cells Injury by Inducing Autophagy and Inhibiting Apoptosis via Regulation of SIRT1.

Background: Oxidative stress-induced endothelial cell death, such as apoptosis and autophagy, plays a critical role in ischemia-reperfusion injury. Protocatechualdehyde (PCA) is a major bioactive component of the traditional Chinese medicine Salvia miltiorrhiza Bunge (Lamiaceae), and it has been proved to be effective in the prevention and treatment of ischemic cardiovascular and cerebrovascular diseases. However, its role in oxidative stress-induced endothelial cell death and its underlying mechanisms remains unclear. This study aims to investigate the effects and mechanisms of PCA on endothelial cell apoptosis and autophagy induced by oxygen-glucose deprivation/reoxygenation (OGD/R) injury. Methods: After OGD/R induction, human umbilical vein endothelial cells (HUVECs) were treated with different concentrations of PCA. Cell viability, apoptosis, and autophagy were detected by Cell Counting Kit-8 assay, flow cytometry, and monodansylcadaverine assay, respectively. Western blot was applied to explore the effects of PCA on the expression levels of relevant protein factors. Results: The results show that PCA significantly promoted cell survival rate and cell proliferation and enhanced the antioxidant activity in OGD/R-induced HUVECs. PCA inhibited HUVECs apoptosis, as evidenced by decreased expression of cleaved-caspase-3, Bcl2-associated X (BAX), and increased expression of Bcl-2. PCA induced autophagy by reducing the expression of P62 while increasing the expression of Beclin-1 and LC3 II/I. Meanwhile, PCA enhanced the expression of Sirtuin 1 (SIRT1) and suppressed the expression of P53. When SIRT1 was inhibited by selisistat or SIRT1 small-interfering RNA, the anti-apoptotic and pro-autophagy abilities of PCA were attenuated. Conclusion: These results demonstrated that PCA rescued HUVECs from OGD/R-induced injury by promoting autophagy and inhibiting apoptosis through SIRT1 and could be developed as a potential therapeutic agent against ischemic diseases.

Carya cathayensis leaf extract attenuates ectopic fat deposition in liver, abdomen and aortic arch in ovariectomized rats fed a high-fat diet.

BACKGROUND: Carya cathayensis1is a commercially cultivated plant in the Zhejiang Province, China. Its nuts exhibit properties of tonifying kidneys and relieving asthma. There have been a few pharmacological studies addressing the function of the leaves of this plant. Our previous studies on C. cathayensis leaf extract (CCE) showed a significant inhibitory effect on weight gain in mice fed a high-fat diet, particularly in female mice. HYPOTHESIS/PURPOSE: To investigate the biological and molecular mechanisms underlying the regulation of ectopic adipose tissue deposition by CCE in ovariectomized rats fed a high-fat diet. STUDY DESIGN: Female Sprague-Dawley rats were ovariectomized and treated with CCE (50, 100, and 200 mg/kg body weight, oral) or estradiol (1 mg/kg body weight, oral) for 8 weeks. METHODS: CCE was subjected to high-performance liquid chromatography to quantify major components. Body weight gain, abdominal fat coefficient, and aortic arch fat coefficient were determined; serum was collected for biochemical analysis; tissues were collected for histopathological examination, quantitative polymerase chain reaction (Q-PCR), and western blotting. RESULTS: The total flavonoid content was determined to be 57.30% in the CCE and comprised chrysin, cardamomin, pinostrobin chalcone, and pinocembrin. Compared with the model group (OVX), CCE treatment reduced body weight gain, abdominal and aortic arch fat coefficients, serum and hepatic lipid profiles, including total cholesterol (TC), total triglycerides (TG), and free fatty acids (FFA) levels; decreased lipid droplets in liver cells; decreased fat accumulation in the aortic arch blood vessel wall and increased its smoothness; decreased the diameter of abdominal fat cells; and reduced serum leptin and adiponectin levels significantly. Serum adiponectin levels significantly correlated with serum TG and hepatic TC levels. Leptin levels positively correlated with serum TG levels and negatively correlated with hepatic TG. Leptin mRNA, peroxisome proliferator-activated receptor (PPARγ) mRNA, and protein expression levels in abdominal adipose tissue were significantly down-regulated. Adiponectin mRNA levels were slightly reduced but not significantly. CONCLUSION: CCE attenuated ectopic fat deposition induced by deficient estrogen and a high-fat diet in rats; this may be associated with activated leptin sensitivity, improved leptin resistance, and regulated adiponectin levels. CCE may improve adipose function to regulate adipocyte differentiation by down-regulating PPARγ. Overall, these results suggest that CCE is a potential phytoestrogen.