Purification and characterization of growth stimulating diadenosine triphosphate from porcine myocardial tissue

The proliferation of vascular smooth muscle cells (VSMC) plays a role in the maintenance of hypertension and atherosclerosis. Diadenosine polyphosphates have been identified as modulators of the cardiovascular system. We detected the existence of diadenosine polyphosphates in the heart and studied their growth stimulating effect on VSMC. Porcine cardiac tissue was deproteinated with perchloric acid and concentrated with a preparative reversed phase C18 column. Then the extract was separated with a size exclusion column, an anion exchange column, and an affinity column. It was then chromatographed with an anion exchange HPLC and a reversed phase HPLC and a purified substance was obtained. Med. Klinik I, Nephrol. Labor, Univ. Klinik Marien hospital, University of Bochum, Herne, Germany (Luo JK, Wang H, Schlüter H, JankowskifJ, Potthoff W, Tepel M and Zidek W) Using the BrdU method, this substance was shown to have a growth stimulating effect that is dose-dependent on VSMC. Matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) analysis showed that the molecular mass of the substance was 757.0, the same as diadenosine triphosphate (AP 3A). UV spectrum indicated the maximum absorption at 259 nm, similar to that of adenosine. Postsource decay (PSD) MALDI MS showed that the substance contained adenine, adenosine, AMP, ADP, and ATP, respectively. Enzymatic cleavage revealed that phosphates in this molecule connected adenosine in 5' position. It was determined from the above data that the substance was Ap 3A. In this study the existence of Ap 3A was demonstrated in the heart. It has a growth stimulating effect on VSMC.

Effect and mechanism of knockout RhoE on the protein expression profiles in cardiac tissues of diabetic rats based on 4D proteomics

Objective:To analyze the effect of RhoE gene expression change on protein expression profiles in the cardiac tissue of diabetic rats,and analyse the possible underlying regulatory mechanism.Methods:Six-week-old male RhoE gene knockout(KO)and wild-type(WT)Sprague Dawley rats were injected intraperitoneally with streptozotocin(70 mg/kg)to establish the type 1 diabetes model(T1DM),with injection of the same amount of sodium citrate saline as the control group.A week later,the fasting blood glucose of the rats was measured daily,and blood glucose concentration 16.7 mmol/L was regarded as a successful model.Two additional weeks later,the hearts of the rats in each group were removed and fourdimensional label-free quantitative proteomics technology(4D-LFQ)was used to analyze the changes of protein profiles in the heart tissue.The related functions,enrichment signals,and protein‑protein interaction network(PPI)of the differentially expressed proteins were analyzed.Results:T1DM rat models were successfully established.Taking a fold change>1.5 and P<0.05 as the threshold,a total of 2931 quantifiable proteins were identified.In the non-diabetic state,the KO group had 26 and 45 significantly up-and downregulated proteins,respectively,compared with the WT group;in the diabetic state,the KO group had 19 and 28 significantly up-and downregulated proteins,respectively,compared with the WT group.The GO annotation results showed that most of the differentially expressed proteins were located in the extracellular matrix,and their biological functions were mainly concentrated in the immune response and energy metabolism.KEGG analysis showed that the signaling pathways associated with the differentially expressed proteins in cardiac tissue after RhoE knockout were mainly related to ribosomes and fat digestion and absorption.Protein interaction network analysis showed that in the cardiac tissue of the KO group,there were more Col1α1-and Col1α2-interacting proteins among the upregulated proteins,and among the down-regulated proteins,related proteins involved in the ribosomal pathway interact more in the network.Conclusion:RhoE knockout significantly changes the protein expression profiles in diabetic cardiac tissue,affecting multiple signaling pathways closely related to diabetic cardiomyopathy.The results provide insights into the pathogenesis and therapeutic target screening of diabetic cardiomyopathy.

Effect of Ganoderma lucidum Polysaccharide on Antioxidative Ability of Rats with Myocardial Injury Induced by Doxorubicin

Objective: The objective is to investigate the effect of Ganoderma lucidum polysaccharide (GLPS) on the oxidative stress induced by doxorubicin (DOX) in cardiomyocytes. Methods: SD rats were divided into control group, DOX group, GLPS low dose and high dose + DOX group. SOD and MDA levels in myocardial tissue were detected in each group. The expression of related proteins in each group was detected by Western blot. Results: GLPS can increase the SOD level and decrease MDA caused by DOX (p Conclusion: Ganoderma polysaccharide can improve the oxidative stress injury of myocardial tissue induced by doxorubicin and play a protective role in myocardial tissue.

Impact of Toll-like Receptor 4 Deficiency on Cerebrocardiac Syndrome

In order to investigate the role of Toll-like receptor 4 （TLR4） in cerebrocardiac syndrome （CCS）, the partial cerebral ischemia/reperfusion （I/R） models in mice with different TLR4 genotypes were established in the present study. TLR4 wild-type （C3H/HeN） and mutant （C3H/HeJ） mice of 6-8 weeks of age were divided into 4 groups at random： C3H/HeN sham group （n=10）, C3H/HeJ sham group （n=10）, C3H/HeN model group （n=10） and C3H/HeJ model group （n=10）. Partial cerebral I/R was caused by the middle cerebral artery occlusion （MCAO） to duplicate CCS models in mice. After the operation, the electrocardiogram （ECG）, the level of tumor necrosis factor-alpha （TNF-c0 in myocardial tissue and the cardiac pathological changes were observed in each group. It was shown that the brain infarct volume in C3H/HeN model group was larger than that in C3H/HeJ model group （P〈0.01）. The ST segment change and T wave inversion occurred frequently in model groups. Moreover, the TNF-ct level in C3H/HeN model group was higher than that in C3H/HeJ model group （P〈0.01）. The myocar- dial injury was aggravated in C3H/HeN group as compared with C3H/HeJ group. It was concluded that TLR4 was implicated in the development of CCS.