Effect of Lentivirus-Mediated miR-499a-3p on Human Umbilical Vein Endothelial Cells.

OBJECTIVE: To explore the possible role of miR-499a-3p in the function of primary human umbilical vein endothelial cells (HUVECs) and the expression of ADAM10 in primary HUVEC. METHOD: miR-499a-3p was first transfected into primary HUVECs via lentivirus vector. The viability, proliferation, and migration of stable transfected primary HUVEC were then determined by flow cytometry, CCK8 assays, scratch tests, and Transwell tests. The transcription of miR-499a-3p and ADAM10 was examined by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of ADAM10 was examined by Western blot (WB). RESULTS: After transfection, miR-499a-3p transcription was significantly increased (P < 0.01), compared to the blank and nonspecific control (NC) groups, while both ADAM10 transcription and expression were significantly decreased (P < 0.05). In contrast, in the inhibitors group, miR-499a-3p transcription was significantly reduced (P < 0.05) whereas both ADAM10 transcription and expression were significantly increased (P < 0.05). The viability, proliferation, and migration of primary HUVECs were significantly impaired (P < 0.05) by the transfection of miR-499a-3p but enhanced by miR-499a-3p inhibitors (P < 0.05). CONCLUSIONS: Upregulation of miR-499a-3p transcription will inhibit the expression of ADAM10 in HUVECs; cell migration and proliferation, however, promote apoptosis. And reverse effects were established by downregulation of miR-499a-3p transcription. All these effects may be achieved by regulating the transcription and expression of ADAM10. These results combined suggested that miR-499a-3p may affect the proliferation, migration, and apoptosis of endothelial cells and regulate AS by regulating ADAM10. miR-499a-3p may become a candidate biomarker for the diagnosis of unstable angina pectoris (UA).

Oridonin Attenuates Myocardial Ischemia/Reperfusion Injury via Downregulating Oxidative Stress and NLRP3 Inflammasome Pathway in Mice.

Oridonin (ORI), the major pharmacological component extracted from a traditional Chinese medicine, possesses a beneficial effect on myocardial ischemia/reperfusion (I/R) injury. However, the underlying molecular mechanism by which ORI effects take place is not completely known. Thus, whether ORI works via downregulating oxidative stress and nod-like receptor protein-3 (NLRP3) inflammasome pathway was investigated in this study. Mice underwent surgery to induce myocardial I/R injury, and some were administered ORI (10 mg/kg/day) pretreatment, while others were not. The myocardial enzymes' levels, infarct area, and inflammatory injury were measured. The activation situation of oxidative stress and NLRP3 inflammasome was also detected. We found that ORI pretreatment significantly alleviated CK-MB and cTnI levels and infarct size induced by I/R. ORI mitigated the inflammatory injury by decreasing the pathological damage and lowering TNF-α, IL-1ß, and IL-18 levels. Moreover, the SOD1 and eNOS levels were significantly increased by ORI, while MDA and iNOS levels were relatively decreased. The oxidative stress was reversed using ORI pretreatment. Importantly, NLRP3 inflammasome pathway was also inhibited by ORI, as reflected by the lower protein levels of NLRP3, caspase-1, and IL-1ß. In conclusion, ORI alleviates myocardial injury induced by I/R via inhibiting the oxidative stress and NLRP3 inflammasome pathway.

Inhibition of miR­155­5p attenuates the valvular damage induced by rheumatic heart disease.

Autoimmunity is involved in the valvular damage caused by rheumatic heart disease (RHD). Increased evidence has linked microRNAs (miRNAs/miRs) to autoimmune disease. Signal transducer and activator of transcription 3 (STAT3) and sphingosine­1­phosphate receptor 1 (S1PR1) and suppressor of cytokine signaling 1 (SOCS1) have been widely studied for their roles in autoimmunity and inflammation. Thus, the current study aims to investigate the role played by miR­155­5p in RHD­induced valvular damage via the S1PR1, SOCS1/STAT3 and interleukin (IL)­6/STAT3 signaling pathways. An RHD rat model was induced by inactivated Group A streptococci and complete Freund's adjuvant. A recombinant adeno­associated virus (AAV­miR155­inhibitor) was used to inhibit the expression of miR­155­5p in the heart. Inflammation and fibrosis were assessed by hematoxylin and eosin staining and Sirius red staining. The expression of miR­155­5p in valvular tissues and serum exosomes was detected by reverse transcription­quantitative PCR. S1PR1, SOCS1, STAT3, phosphorylated STAT3, IL­6 and IL­17 protein expression was detected by western blotting and immunohistochemistry. The relationships between miR­155­5p and S1PR1 and SOCS1 were detected by dual luciferase assays. Cytokine concentrations were measured by ELISA. The expression of miR­155­5p in valve tissues and serum exosomes was increased along with decreased S1PR1 and activated SOCS1/STAT3 signaling in the RHD model. The expression of IL­6 and IL­17 was increased in the valves and the serum. Dual luciferase assays showed that miR­155­5p directly targeted S1PR1 and SOCS1. Inhibition of valvular miR­155­5p through AAV pretreatment increased S1PR1 expression and inhibited activation of the SOCS1/STAT3 signal pathway as a result of attenuated valvular inflammation and fibrosis as well as a decrease in IL­6 and IL­17 in the valves and serum. These results suggest that inhibition of miR­155­5p can reduce RHD­induced valvular damage via the S1PR1, SOCS1/STAT3 and IL­6/STAT3 signaling pathways.

Psychometric properties of the Chinese version of the empathy quotient among Chinese minority college students.

BACKGROUND: When the minority college students from the ethnic minority communities come to study in Chinese Han region, they encounter adapting difficulties of culture and socio-psychology, in which empathy plays a crucial role. Current instruments used to measure empathy have many limited effectiveness. The empathy quotient (EQ) scale which has been validated in many countries was explicitly designed for clinical applications and was intended to be sensitive to a lack of empathy. This study is to develop a complete Chinese version of the EQ scale and to assess its reliability and validity among Chinese minority college students in the Han Chinese region. METHODS: A total of 1638 Chinese minority college students in the Han region were selected and were randomly divided into two groups. One group of 818 students took part in the implementation of the exploratory factor analysis while the other group of 820 students participated in the confirmatory factor analysis. RESULTS: Twenty-nine items of the EQ were retained based on the factor analysis and four factors were extracted: self-awareness, cognitive empathy, social skills, and emotional reactivity, which can explain 51.793% of the total variance. The factors of the EQ scale were significantly correlated with each other, with the correlation coefficient ranging from 0.316 to 0.563. The coefficient of internal consistency (Cronbach's α) was 0.824 for the total scale and ranged from 0.640 to 0.818 for the subscales. Confirmatory factor analysis proved that the measured data fitted well with the hypothesized four-factor model. All of the items in the scale fitted the model well, and the point-measure correlation coefficient had acceptable consistency. CONCLUSIONS: The refined 29-item Chinese version of the EQ possesses good reliability and validity, and can be applied in assessing empathy among Chinese minority college students.

Effects of mTOR/NF-κB signaling pathway and high thoracic epidural anesthesia on myocardial ischemia-reperfusion injury via autophagy in rats.

We investigated the role of mammalian target of rapamycin/nuclear factor-kappa B (mTOR/NF-κB) signaling pathway in high thoracic epidural anesthesia (HTEA) against myocardial ischemia-reperfusion (I/R) injury in rats. The rat model of myocardial I/R injury was established. Ninety rats were divided into the normal, sham, I/R, eHTEA, the PDTC, and HTEA + PDTC groups. ELISA was applied to detect cardiac function indexes. HE staining was conducted to observe histopathological changes of myocardial tissues, and TTC staining was performed to detect the myocardial infarction size. TUNEL staining was adopted to detect the cell apoptosis rate. The mRNA and protein levels of mTOR, NF-κB, Fasl, Bcl-2 and Bax, and LC3-I, LC3-II, BNIP3, and Atg5 were detected by RT-qPCR and Western blotting, respectively. The findings indicated that compared with the normal and sham groups, the I/R, PDTC, and HTEA groups showed the larger myocardial infarction size and increased cell apoptosis rate, while the results in the HTEA + PDTC group were opposite. Compared with the normal and sham groups, the I/R group showed reduced mRNA and protein levels of Bcl-2, LC3, BNIP3, and Atg5, and elevated mRNA and protein levels of mTOR, p50, p65, Bax, and Fasl, while the HTEA + PDTC group revealed the opposite results, and the PDTC and HTEA group revealed the increased mRNA and protein levels of Bcl-2, LC3, BNIP3, Atg5, mTOR, p50, p65, Bax, and Fasl. These results prove that the inhibition of mTOR/NF-κB signaling pathway potentiates HTEA against myocardial IR injury by autophagy and apoptosis in rats.