[Effects of maslinic acid on isoproterenol-induced myocardial fibrosis in mice].

Objective: To investigate the effect of maslinic acid (MA) on isoproterenol (ISO)-induced myocardial fibrosis in mice. Methods: ISO was used to induce myocardial fibrosis in adult male C57BL/6 mice, and MA was administered for two weeks to detect the effects of MA on cardiac function and fibrosis. Molecular changes of fibrosis markers and signaling pathways were detected by RT-PCR and western blotting. Phosphate buffer saline (PBS), PBS+SB203580 (p38 MAPK inhibitor), PBS+MA, ISO, ISO+SB203580, ISO+MA were added to the primary cultured rat fibroblasts. Cells were collected after 48 h for subsequent detection. Results: In this study, the mouse model of myocardial fibrosis was successfully established. The left ventricular faction shortening (FS) and maximum rate of rise and maximum rate of fall of pressure in left ventricular chamber (±dp/dt) of the ISO+MA group were significantly higher than those of the ISO group ((35.1±1.8)% vs (28.5±2.6)%, (7 256±153) mmHg/s vs (6 402±240) mmHg/s, (7 156±163) mmHg/s vs (6 319±219) mmHg/s, all P<0.05). The levels of interstitial and perivascular collagen deposition in the ISO+MA group were higher than those in the ISO group (P<0.05), the relative mRNA levels of COL-1, COL-3 and TGF-ß in the ISO+MA group were significantly lower than those in the ISO group, with the relative expression levels of 1.70±0.24 vs 3.69±0.34, 1.72±0.56 vs 4.84±0.82, 1.52±0.19 vs 2.64±0.29, respectively (all P<0.05). The phosphorylation levels of p38 MAPK, Smad3 and protein expression level of TGF-ß1 in ISO+MA group were lower than those in ISO group (relative expression levels were 1.67±0.35 vs 2.61±0.58, 1.68±0.23 vs 2.52±0.19,1.56±0.15 vs 2.48±0.26, respectively, all P<0.05). The results of in vitro cell experiments showed that the mRNA levels of COL-1, COL-3 and TGF-ß in the SB203580 and MA groups were significantly lower than those in the ISO group (relative expression levels were 2.25±0.51, 2.16±0.48 vs 5.29±1.21; 1.58±0.34, 1.69±0.29 vs 4.97±1.32; 1.41±0.31, 1.55±0.38 vs 3.53±0.56, respectively, all P<0.05). The phosphorylation levels of p38 MAPK and Smad3 in the SB203580 MA groups was significantly lower than those in the ISO group, and the protein expression level of TGF-ß1 was lower than that in the ISO group (1.81±0.18, 1.77±0.16 vs 2.56±0.32; 1.85±0.21, 1.81±0.17 vs 2.48±0.37; 1.84±0.24, 1.72±0.17 vs 2.52±0.29, all P<0.05). Conclusion: Maslinic acid can inhibit the phosphorylation of p38 MAPK, thereby preventing the canonical TGF-ß1/Smads fibrosis signaling pathway to achieve an anti-fibrosis role.

[The effect and mechanism of heat shock protein 47 on streptozotocin-induced diabetic cardiomyopathy].

Objective: To investigate the effect and specific mechanism of heat shock protein 47 (HSP47) on streptozotocin (STZ)-induced diabetic cardiomyopathy (DCM). Methods: A mouse model of type 1 diabetic cardiomyopathy was established by intraperitoneal injection of STZ. After 8 weeks of successful modeling, HSP47 was overexpressed by tail vein injection, and the heart of the mouse was harvested after 6 weeks. Hematoxylin-eosin (HE) staining and Sirius red (PSR) staining were used to detect the cross-sectional area of myocardial cells and myocardial fibrosis, respectively. Immunofluorescence staining with α-smooth muscle actin (α-SMA) and collagen Ⅰ was used to detect the degree of fibrosis activation. The expression level of fibrosis-related proteins was determined by Western blot. Results: The expression level of HSP47 in the myocardium of the diabetic group up-regulated (2.014±0.264 vs 1.004±0.064, P<0.001). The area of myocardial cells in the diabetic group was increased compared with the control group [(235.3±20.7) µm(2) vs (172.8±13.6) µm(2), P<0.001] and the cross-sectional area of myocardial cells in the HSP47 overexpression-diabetes group was further increased [(302.2±41.0) µm(2) vs (235.3±20.7) µm(2), P=0.009], while the mRNA levels of mouse cardiac hypertrophic markers atrial natriuretic peptide (ANP), type B brain natriuretic peptide (BNP), myosin heavy chain ß (ß-MHC) further upregulated (all P<0.001). Compared with the control group, the myocardial fibrosis content in the diabetic group increased [(7.333±1.127)% vs (4.837±0.775)%, P=0.002] and the left ventricular fibrosis content of the HSP47 overexpressing diabetic group further increased [(9.175±1.008)% vs (7.333±1.127)%, P=0.025] and the mRNA levels of fibrosis index collagenⅠ, collagen Ⅲ, connective tissue growth factor (CTGF) and transforming growth factor ß (TGFß) further up-regulated (all P<0.001). Immunofluorescence results showed that compared with the control group, the expression of collagenⅠ up-regulated in the endothelial stroma of the diabetic group and the content of collagenⅠ in the HSP47 over-expressing diabetic group was higher (P<0.001). Western blot results indicated that the phosphorylation level of Smad3 and the protein levels of α-SMA and TGFß in HSP47 overexpressing diabetic group increased, compared with those of diabetic group (all P<0.001). Conclusion: HSP47 ameliorates STZ-induced diabetic myocardial fibrosis by activating the TGFß/Smad3 signaling pathway.

[Cinnamaldehyde attenuates pressure overload-induced cardiac fibrosis via inhibition of endothelial mesenchymal transition].

Objective: The aim of this study is to investigate the underlying mechanism of cinnamaldehyde attenuating pressure overload-induced cardiac fibrosis. Methods: The mice were randomly divided into control group, model group and treatment group by random number table and each group had 8 mice.Cardiac hypertrophy was induced by aortic banding. Heart vascular density was detected by immunohistochemical staining of CD31.The expression level of stromal cells marker α-smooth muscle actin (α-SMA) was detected by immunofluorescence staining in different groups.The expression levels of endothelial cell associated markers and stromal cell associated markers were detected by using Western blotting.The possible molecular pathway was also screened by using Western blotting. Human umbilical vein endothelial cell (HUVECs) were stimulated with TGFß1 and cultured with 10 nmol/L cinnamomum for 24 hour to further confirm the mechanism. Results: Eight weeks after operation, the vascular density was significantly decreased in model group mice heart.The expressions of stromal cells markers were increased (α-SMA: 2.57±0.38; Vimentin: 0.58±0.02) and endothelial cell markers were reduced (CD31: 0.58±0.29; CD34: 0.62±0.21). While cinnamicaldehyde treatment significantly increased the mouse heart vascular density, increased endothelial cell markers expression (CD31: 1.51±0.11; CD34: 2.37±0.44; P<0.05), and reduced stromal cells marker expression (α-SMA: 1.22±0.14; Vimentin: 0.35±0.03; P<0.05). Further studies showed that the anti-fibrosis effect of cinnamicaldehyde was mainly through the TGFß /smad signaling pathway.10 nmol/L cinnamomum attenuated TGFß1 induced endothelial mesenchymal transition in HUVECs. Conclusion: Cinnamaldehyde may be able to retard the progression of cardiac fibrosis, via blocking endothelial to mesenchymal transition, which, in verse, is through regulating TGFß /smad signaling pathway.

Effectiveness of microsatellite and single nucleotide polymorphism markers for parentage analysis in European domestic pigs.

Parentage analysis and individual identification are recent, promising methods that have been applied to evolutionary and ecological studies, as well as conservation management. Parental exclusion relying on polymorphic microsatellites has been used worldwide in parentage determination, while the low mutation rate and genotyping error rate of single nucleotide polymorphisms (SNPs) make them another important marker for pedigree tracing. Here, we compared the effectiveness of microsatellites and SNP markers in European pigs. We also measured and presented the minimum and optimal criteria for SNP markers to be used in paternity and identity analysis. Our findings may contribute to the development of techniques for future molecular evolution and conservation studies, as well as breeding programs.

Sulforaphane protects H9c2 cardiomyocytes from angiotensin II-induced hypertrophy.

BACKGROUND: Cardiac hypertrophy is an adaptive process of the heart in response to various stimuli, but sustained cardiac hypertrophy will finally lead to heart failure. Sulforaphane-extracted from cruciferous vegetables of the genus Brassica such as broccoli, brussels sprouts, and cabbage-has been evaluated for its anticarcinogenic and antioxidant effects. AIMS: To investigate the effect of sulforaphane on angiotensin II (Ang II)-induced cardiac hypertrophy in vitro. METHODS: Embryonic rat heart-derived H9c2 cells were co-incubated with sulforaphane and Ang II. The cell surface area and mRNA levels of hypertrophic markers were measured to clarify the effect of sulforaphane on cardiac hypertrophy. The underlying mechanism was further investigated by detecting the activation of Akt and NF-κB signaling pathways. RESULTS: We found that H9c2 cells pretreated with sulforaphane were protected from Ang II-induced hypertrophy. The increasing mRNA levels of ANP, BNP, and ß-MHC in Ang II-stimulated cells were also down-regulated after sulforaphane treatment. Moreover, sulforaphane repressed the Ang II-induced phosphorylation of Akt, GSK3ß, mTOR, eIF4e, as well as of IκBα and NF-κB. CONCLUSION: Based on our results, sulforaphane attenuates Ang II-induced hypertrophy of H9c2 cardiomyocytes mediated by the inhibition of intracellular signaling pathways including Akt and NF-κB.

Troponin T measurement can predict persistent left ventricular dysfunction in peripartum cardiomyopathy.

OBJECTIVE: To determine whether measurement of cardiac troponin T (cTnT) concentration in newly diagnosed peripartum cardiomyopathy (PPCM) can be used to predict persistent left ventricular dysfunction after a 6-month follow-up. PATIENTS AND METHODS: This was a prospective, multiple-centre clinical trial that studied 106 patients with newly diagnosed PPCM surviving over 6 months. cTnT concentration was measured within 2 weeks of the onset of PPCM. RESULTS: Serum cTnT concentration was negatively correlated with left ventricular ejection fraction (LVEF) at follow-up (LVEF, r = -0.518, p = 0.0001). Analysis by receiver operator characteristic curve yielded an area under the curve of 0.764 (95% CI 0.669 to 0.860, p = 0.0001, vs null hypothesis value 0.5) for cTnT, and a cTnT concentration cut off of >0.04 ng/ml, predicting persistent left ventricular dysfunction with a sensitivity of 54.9% and a specificity of 90.9%. Among 106 recruited patients, there were 33 patients with cTnT concentrations >0.04 ng/ml and 73 patients with cTnT concentrations < or =0.04 ng/ml. After a 6-month follow-up, there was significantly smaller LVEF (35.42% (13.04% vs 50.16% (10.48%, p = 0.0001) and more persistent left ventricular dysfunction (84.8% vs 31.5%, OR = 12.17 (95% CI 4.17 to 35.57), p = 0.001) in patients with cTnT >0.04 ng/ml than in patients with cTnT < or =0.04 ng/ml. CONCLUSION: Serum cTnT concentration measured within 2 weeks of the onset of PPCM was correlated negatively with LVEF at follow-up. This marker offers a simple, quick, inexpensive, non-invasive method for predicting a persistent LVEF of < or =50%. A cTnT concentration of >0.04 ng/ml predicted persistent left ventricular dysfunction with a sensitivity of 54.9% and a specificity of 90.9%.

Comparison of rate control and rhythm control in patients with atrial fibrillation after percutaneous mitral balloon valvotomy: a randomised controlled study.

OBJECTIVE: To compare rate control and rhythm control strategies in patients with atrial fibrillation (AF) after percutaneous mitral balloon valvotomy (PMV). METHODS: 183 patients with AF after successful PMV, with AF duration

[Effect of buyang huanwu decoction on platelet function and fibrinolytic activity of unstable angina pectoris patients].

OBJECTIVE: To explore the effect of Buyang Huanwu Decoction (BYHWD) on the platelet function and fibrinolytic activity of unstable angina pectoris patients. METHODS: Using randomized single blind method divided 60 unstable angina pectoris patients into conventional treatment (control) group and conventional plus BYHWD (TCM) group. RESULTS: BYHWD could lower the levels of platelet aggregation, thromboxane B2 and the inhibitor of fibrinolysinogen activator (P < 0.05), but the activity of histo-type of fibrinolysinogen activator was enhanced. CONCLUSION: BYHWD was effective in treating unstable angina pectoris.