Circulating MiR-19b-3p, MiR-134-5p and MiR-186-5p are Promising Novel Biomarkers for Early Diagnosis of Acute Myocardial Infarction.

BACKGROUND/AIMS: Recent studies have shown that circulating microRNAs (miRNAs) are emerging as promising biomarkers for cardiovascular diseases. This study aimed to determine whether miR-19b-3p, miR-134-5p and miR-186-5p can be used as novel indicators for acute myocardial infarction (AMI). METHODS: To investigate the kinetic expression of the three selected miRNAs, we enrolled 18 patients with AMI and 20 matched controls. Plasma samples were collected from each participant, and total RNA was extracted. Quantitative real-time PCR and ELISA assays were used to investigate the expression of circulating miRNAs and cardiac troponin I (cTnI), respectively. Plasma samples from another age- and gender-matched cohort were collected to investigate the impact of medications for AMI on the expression of the selected miRNAs. RESULTS: Levels of plasma miR-19b-3p, miR-134-5p and miR-186-5p were significantly increased in early stage of AMI. Plasma miR-19b-3p and miR-134-5p levels reached peak expression immediately after admission (T0), whereas miR-186-5p achieved peak expression at 4 h after T0. All of these times were earlier than the peak for cTnI (8 h after T0). In addition, all three miRNAs were positively correlated with cTnI. Receiver Operating Characteristic (ROC) analysis indicated that each single miRNA showed considerable diagnostic efficiency for predicting AMI. Furthermore, combining all three miRNAs in a panel increased the efficiency of distinguishing between patients with AMI and controls. Moreover, we found that heparin and medications for AMI did not impact the expression of these circulating miRNAs. CONCLUSION: Circulating miR-19b-3p, miR-134-5p and miR-186-5p could be considered promising novel diagnostic biomarkers for the early phase of AMI.

The mechanism of hyperoside protection of ECV-304 cells against tert-butyl hydroperoxide-induced injury.

The aim of the present study was to investigate the mechanism of hyperoside protecting ECV-304 cells against tertbutyl hydroperoxide (TBHP)-induced injury. ECV-304 cell viability was measured by MTT assay. Cellular morphologic changes were observed using phase contrast microscopy. The genotoxic effects of TBHP and the protective ability of hyperoside were assessed by the Comet test. Lipid peroxidation was measured by HPLC method. The cellular redox status was determined from GSH/GSSG ratios. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. Western blot analysis was used to evaluate the levels of cytochrome c, p53, SIRT1, Bax and Bcl-2 expression. The results showed that 128 mumol/l hyperoside could effectively protect TBHP-treated ECV-304 cells from death, increase superoxide dismutase activity and significantly decrease malondialdehyde production. Hyperoside was effective in protecting against the induction of oxidized DNA bases and redox state alterations induced by TBHP. Furthermore, the release of proapoptotic cytochrome c from mitochondria was reduced by hyperoside, which increased the expression of antiapoptotic SIRT1 and inhibited the translocation of Bax from cytoplasm to mitochondria. Taken together, these results indicate that hyperoside is effective in protecting against the oxidative damage induced by TBHP. The mechanism of hyperoside protecting against ECV-304 cell apoptosis by TBHP is related with resuming mitochondrial function and regulating the expression of SIRT1 and Bcl-2 family members.

Magnolol-induced H460 cells death via autophagy but not apoptosis.

We have reported that the protective effect of Magnolol on TBHP-induced injury in human nonsmall lung cancer H460 cells is partially via a p53 dependent mechanism. In this study, we found that Magnolol displayed a stimulatory effect at low concentrations (< or = 20 microM) whilst inhibitory effect at high concentrations (> or = 40 microM) in H460 cells. To investigate the mechanism of inducing the biphasic effect in H460 cells with Magnolol, we showed that Magnolol stimulated DNA synthesis at low concentrations and displayed an inhibition effect at high concentrations in H460 cells. More importantly, the inhibition of DNA synthesis was accompanied by the S phase cell cycle arrest and the appearance of intense intracytoplasmic vacuoles. These vacuoles can be labeled by autophagic marker monodansylcadaverin (MDC), 3-methyladenine (3-MA), an inhibitor of autophagy, was able to inhibit the occurrence of autophagy. The results of the LDH activity assay and TUNEL assay also showed that Magnolol at high concentrations inhibiting H460 cell growth was not via apoptotic pathway. Furthermore, accompanied by the occurrence of autophagy, the expression of phospho-Akt was down-regulated but PTEN significantly was up-regulated. In conclusion, Magnolol induces H460 cells death by autophagy but not apoptotic pathway. Blockade of PI3K/PTEN/Akt pathway is maybe related to Magnolol-induced autophagy. Autophagic cells death induction by Magnolol underlines the potential utility of its induction as a new cancer treatment modality.

[Effects of acupoint area and non-acupoint area of eye-acupuncture on expressions of VIP and AQP 8 in colonic tissues in rats with D-IBS].

OBJECTIVE: To explore the point specificity of eye-acupuncture and the mechanism of eye-acupuncture on diarrhea-predominant irritable bowel syndrome (D-IBS). METHODS: Forty male Wistar rats of SPF grade were randomly divided into a normal group, a model group, a eye-acupuncture point (AA) group and a non-point (NA) group. The D-IBS rat model was established with the combination methods of the chronic stress and binding limbs. The AA group was treated by acupuncture at "low energizer area", "large intestine area", "liver area" and "spleen area", and the NA group by acupuncture at 3 mm apart from the same points area mentioned above, and the normal group and the model group with no intervention. The rate of feces moisture content was detected. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA of aquaporin 8 (AQP 8) in colon. Protein expressions of the vasoactive intestinal peptide (VIP) and AQP 8 in colon were detected by SABC immunohistochemistry method. RESULTS: Compared with normal group, the rate of feces moisture content at the 18th and 25th days, VIP protein in colon mucosa, myenteric nerve plexus and hypo-mucosa nerve plexus increased significantly (all P < 0.01), and AQP 8 mRNA in colon mucosa decreased significantly in model, AA and NA group (P < 0.05, P < 0.01); AQP 8 protein in colon mucosa decreased significantly in model group and NA group (both P < 0.01). Compared with model group, the rate of feces moisture content at the 25th day and VIP protein in colon mucosa decreased significantly (P < 0.01, P < 0.05), and AQP 8 mRNA and protein increased significantly (P < 0.05, P < 0.01) in AA group. Compared with AA group, the rate of feces moisture content at the 25th day and VIP protein in colon mucosa increased significantly (both P < 0.01), and AQP 8 mRNA and protein decreased significantly (both P < 0.01) in NA group. CONCLUSION: Eye-acupuncture has a good therapeutic effect on D-IBS. It is suggested that one of the mechanism is relate to increase AQP 8 in colon tissue and restrain the expression of VIP. Non-point area of eye-acupuncture has no obviously therapeutic effect and so to illustrate the point specificity of eye-acupuncture.

[Effects of eye-acupuncture therapy on the expression of AQP4 in brain tissue of rats with acute cerebral ischemia-reperfusion injury].

OBJECTIVE: To explore the mechanism of the eye-acupuncture for treatment of acute cerebral ischemia-reperfusion injury. METHODS: Thirty-two healthy SD rats were randomly divided into a normal group, a sham operation group, a model group and an eye-acupuncture group, 8 rats in each group. The rat model of cerebral ischemia-reperfusion was established with thread occlusion method in the model group and the eye-acupuncture group. The eye-acupuncture group was treated by eye-acupuncture at "liver region", "upper energizer area", "lower energizer area" and "kidney region" for 20 min immediately after reperfusion and at 30 min before sampling. No treatment was done in the normal group and the sham operation group, and no thread occlusion was performed in the sham operation group. The Neurologic impairment was scored and the methods of immunohistochemistry staining, western-blotting and real-time fluorescent quantitation polymerase chain reaction (RQ-PCR) were taken to detect the expression of the aquaporin protein 4 (AQP4) and its mRNA in cerebral cortex after reperfusion for 3 hours. RESULTS: The neurologic impairment score of 1.50 +/- 0.54 in the eye-acupuncture group was significant lower than 2.63 +/- 0.92 in the model group (P < 0.01). The expression of the AQP4 protein by immunohistochemistry and western-blot respectively were 116.33 +/- 10.24 and 0.53 +/- 0.04 in the normal group, 118.97 +/- 12.72 and 0.55 +/- 0.07 in the sham operation group, and 129.30 +/- 18.36 and 0.67 +/- 0.08 in the eye-acupuncture group, with statistical significance compared to 150.88 +/- 15.82 and 0.94 +/- 0.04 in the model group (all P < 0.01), and there were significant differences between the eye-acupuncture group and the normal group (both P < 0.01). The tendency in the expression of AQP4 protein and its mRNA in all the group were almost the same. CONCLUSION: The eye-acupuncture therapy can relieve the cerebral ischemia-reperfusion injury and the protective mechanism is related to the downregulation of the cerebral AQP4 expression.

[Effect of eye-acupuncture on cerebral intracellular adhesion molecule-1 expression in rats with acute cerebral ischemia-reperfusion injury].

OBJECTIVE: To investigate the effect of the eye-acupuncture therapy on cerebral intercellular adhesion molecule-1 (ICAM-1) expression in rats with acute cerebral ischemia-reperfusion injury (CI/RI) so as to reveal its underlying mechanism. METHODS: Forty male SD rats were randomly and equally divided into normal, sham operation (sham), model and eye-acupuncture groups. Acute CI/RI model was established by occlusion of the middle cerebral artery with a modified suture-blocking method. Filiform needles were inserted into the "Qan" (Liver Area), "Shangjiao" (Upper Energizer Area), "Xiajiao" (Lower Energizer Area), etc. of the eye-acupuncture acupoints, and retained for 20 min. The treatment was conducted twice daily, 7 times altogether. Seventy-two hours after Acute CI/RI, the neurological behavior was assessed by ZeaLonga neurophysical impairment scale. Cerebral ICAM-1 protein and ICAM-1 mRNA expression levels were detected by Western blot (Penumbra area of the right cerebral infarction region) and real time-polymerase chain reaction (PCR) methods, respectively. RESULTS: After modeling, the neurophysical impairment score of the model group was increased from 0 to (2.63 +/- 0.92) points on the first day and (2.63 +/- 0.74) points 72 h following Acute CI/RI. Correspondingly, cerebral ICAM-1 protein and mRNA expression was up-regulated considerably in the model group than in the normal control group (P < 0. 01). In comparison with the model group, the neurologic impairment score, and cerebral ICAM-1 protein and mRNA expression levels of the eye-acupuncture group were down-regulated obviously (P < 0.01, P < 0.05). No significant differences were found between the normal control and sham groups in neurologic impairment score and the expression levels of the ICAM-1 protein and ICAM-1 mRNA(P > 0.05). CONCLUSION: Eye-acupuncture therapy can improve the cerebral ischemia-reperfusion injury induced behavior changes, which may be related to its effects in down-regulating the expression of cerebral ICAM-1 protein and gene.

[Effects of eye-acupuncture therapy on serum and colonic SP and VIP contents in rats with irritable bowel syndrome].

OBJECTIVE: To observe the effect of the eye-acupuncture therapy on serum and colonic substance P (SP) and vasoactive intestinal peptide (VIP) contents in rats with irritable bowel syndrome (IBS) so as to explore its underlying mechanism. METHODS: Forty male Wistar rats were equally randomized into control group, IBS model group, eye-acupuncture group and medication (Pinaverium bromide, 7.5 mg/kg, twice daily, intragastric administration) group. IBS model was established by giving the rat with chronic stress stimulation (cold-water swimming, tail clamping, electrical shock, etc.) for 18 days. Eye-acupuncture of Xiajiao (Low Energizer) Area, Pi (Spleen) Area, Gan (Liver) Area and Dachang (Large Intestine) Area was given to the rat 20 min, twice daily for 7 d. Histopathological changes of the colon tissue were displayed by HE staining; and serum and colonic SP and VIP contents were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: No significant difference was found among 4 groups in the histopathological changes of the colon. In comparison with normal control group, both serum and colonic SP and VIP contents in model group increased significantly (P < 0.01), while compared with model group, those in eye-acupuncture and medication groups lowered considerably (P < 0.01). CONCLUSION: Eye-acupuncture can reduce serum and coIonic SP and VIP contents in IBS rats, which may play a role in relieving IBS in eye-acupuncture clinic.