Animals ; Cardiovascular System ; chemistry ; Female ; Group II Phospholipases A2 ; blood ; Group IV Phospholipases A2 ; blood ; Group V Phospholipases A2 ; blood ; Group X Phospholipases A2 ; blood ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

Phospholipase C (PLC) is a key enzyme in phosphatidyl inositol turnover during signal transduction. The 12 mammalian PLC isozymes identified to date can be divided into five subtypes, beta-type, gamma-type, delta-type, epsilon-type and zeta-type, with extensive difference in structure, regulatory mechanism and tissue distribution. PLC plays important roles not only in sperm acrosome reaction but also in egg activation. The present studies are reviewed on the structure, regulation and function of PLC, especially its function in male reproduction, including triggering Ca2+ oscillations in eggs to activate the eggs and helping embryo development. And the prospect of the clinical application of PLC is discussed.
Acrosome Reaction ; Animals ; Calcium Signaling ; Humans ; Reproduction ; physiology ; Signal Transduction ; Type C Phospholipases ; chemistry ; physiology

OBJECTIVESTo establish and evaluate the anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb).

METHODSAfter having been separated and purified from human seminal plasma by PEG precipitation, Sephacryl S-300 column chromatography, DEAE-Sephadex A-25 column chromatography and HA column chromatography, PLA2 was regarded as an antigen to immune BALB/C mouse to produce anti-human seminal plasma PLA2 McAb. The PLA2 McAb sensitivity and specificity were performed by ELISA technique and Western-blot analysis, respectively.

RESULTSThe molecular weight of PLA2 depurated with 245 fold purification from human seminal plasma was about 34,900, while the sensitivity and typing of its McAb were 1:5(6)-1:5(8) and IgM (kappa) with a satisfied Western-Blot results.

CONCLUSIONSThe PLA2, which had not been reported in international and domestic papers, may be a new type of PLA2. The establish of its McAb will provide significant tools for the research of the relationship between PLA2 in human seminal plasma and male fertility.

Antibodies, Monoclonal ; immunology ; DEAE-Dextran ; analogs & derivatives ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Humans ; Molecular Weight ; Phospholipases A ; immunology ; isolation & purification ; metabolism ; Phospholipases A2 ; Semen ; enzymology

The storage and transportation of raw milk at low temperatures promote the growth of psychrotrophic bacteria and the production of thermo-stable enzymes, which pose great threats to the quality and shelf-life of dairy products. Though many studies have been carried out on the spoilage potential of psychrotrophic bacteria and the thermo-stabilities of the enzymes they produce, further detailed studies are needed to devise an effective strategy to avoid dairy spoilage. The purpose of this study was to explore the spoilage potential of psychrotrophic bacteria from Chinese raw milk samples at both room temperature (28 °C) and refrigerated temperature (7 °C). Species of Yersinia, Pseudomonas, Serratia, and Chryseobacterium showed high proteolytic activity. The highest proteolytic activity was shown by Yersinia intermedia followed by Pseudomonas fluorescens (d). Lipolytic activity was high in isolates of Acinetobacter, and the highest in Acinetobacter guillouiae. Certain isolates showed positive β-galactosidase and phospholipase activity. Strains belonging to the same species sometimes showed markedly different phenotypic characteristics. Proteases and lipases produced by psychrotrophic bacteria retained activity after heat treatment at 70, 80, or 90 °C, and proteases appeared to be more heat-stable than lipases. For these reasons, thermo-stable spoilage enzymes produced by a high number of psychrotrophic bacterial isolates from raw milk are of major concern to the dairy industry. The results of this study provide valuable data about the spoilage potential of bacterial strains in raw milk and the thermal resistance of the enzymes they produce.
Animals ; Bacteria/genetics* ; Bacterial Proteins/chemistry* ; Biofilms ; Cold Temperature ; Dairy Products ; Endopeptidases/chemistry* ; Enzyme Stability ; Food Microbiology ; Hot Temperature ; Lipase/chemistry* ; Milk/microbiology* ; Peptide Hydrolases/chemistry* ; Phospholipases/chemistry* ; RNA, Ribosomal, 16S/genetics* ; Raw Foods/microbiology* ; beta-Galactosidase/chemistry*

OBJECTIVETo investigate the therapeutic mechanism of rhubarb in protecting the intestinal muco-membranous barrier in the mice.

METHODBal b/c mice were divided into 2 groups, gavaged with normal saline and 10% rhubarb decoction, respectively. The animals were killed after 24 hours after the treatments. The intestinal juice was collected after intestinal lavage and centrifuged for determination of IgA, total protein, C3, high density lipoprotein, type II PLA2 activity, and content of lysozyme. At the same time, 40 mg of small intestine were incised in each mouse. Reverse transcription polymerase chain reaction (RT-PCR) and gel image analysis were performed to detect the content of the cryptdin gene expression.

RESULTThe content of IgA, total protein, the C3, lysozyme, and the type II PLA2 activity in intestinal lavaged juice exhibited the statistical differences between the two groups (P < 0.05). There were no significant difference in the ontents of HDL, cryptdin-1 and cryptdin-4 gene expression between the two groups (P > 0.05).

CONCLUSIONRhubarb could increase secretion of several immune associated substances of the mucous membrane in normal intestine, indicating a possibility to abate the injury of intestine mucus resulted from severe stress induced by trauma, burn and shock. Through above mechanisms Rhubarb may also reduce the incidence of bacterial translocation and systemic inflammatory reaction syndrome (SIRS).

Animals ; Complement C3 ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Group II Phospholipases A2 ; Immunoglobulin A ; metabolism ; Intestinal Mucosa ; metabolism ; Intestinal Secretions ; metabolism ; Intestine, Small ; metabolism ; Mice ; Mice, Inbred BALB C ; Muramidase ; metabolism ; Phospholipases A ; metabolism ; Phospholipases A2 ; Plants, Medicinal ; chemistry ; Proteins ; metabolism ; Rheum ; chemistry

This study was to report the effect of Tangshen Formula on phospholipids metabolism in diabetic nephropathy patients. A normal phase-HPLC-TOF/MS method was used in this study for the determination of seven species of phospholipids in human plasma. Then, the concentration changes of potential phospholipids biomarkers were discussed in diabetic nephropathy phase III and phase IV patients among different groups, including before and 3, 6 months after administration of Tangshen Formula. Significant increases of PE750, PI885, PC792, PC826, PC830, PC854 and PC802 levels were observed 6 months after administration of Tangshen Formula and conventional western medicine, as well as a decrease of LPC540 level, when compared with those before medication. Concentrations of all the potential phospholipids biomarkers showed a tendency towards normal levels; however, both the improvement degree and onset time of these compounds were not same. Additionally, Tangshen Formula treatment based on conventional western medicine treatment was more efficient in adjusting the levels of these compounds when compared with western medicine treatment alone, especially for the phase IV patients. These results indicated that Tangshen Formula was capable in regulating and improving phospholipids metabolism in diabetic nephropathy patients, which may be related with the direct or indirect inhibition of protein kinase C pathway and the corresponding reduction of phospholipase A2 activity. Therefore, Tangshen Formula may be used as an effective drug for diabetic nephropathy therapy, at least as an adjunctive therapeutic drug.
Diabetic Nephropathies ; blood ; metabolism ; Double-Blind Method ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Glycerophospholipids ; blood ; Humans ; Lysophosphatidylcholines ; blood ; Phospholipases A2 ; metabolism ; Phospholipids ; blood ; classification ; Plants, Medicinal ; chemistry ; Protein Kinase C ; metabolism ; Signal Transduction ; Sphingomyelins ; blood

OBJECTIVETo investigate the therapeutic effects and mechanisms of Salvia miltiorrhizae (Danshen) in the treatment of severe acute pancreatitis (SAP)- or obstructive jaundice (OJ)-induced heart injury.

METHODSA total of 288 rats were used for SAP- (n=108) and OJ-associated (n=180) experiments. The rats were randomly divided into sham-operated, model control, and Salvia miltiorrhizae-treated groups. According to the difference of time points after operation, SAP rats in each group were subdivided into 3, 6 and 12 h subgroups (n=12), whereas OJ rats were subdivided into 7, 14, 21, and 28 d subgroups (n=15). At the corresponding time points after operation, the mortality rates of the rats, the contents of endotoxin and phospholipase A2 (PLA2) in blood, and pathological changes of the hearts were investigated.

RESULTSThe numbers of dead SAP and OJ rats in the treated groups declined as compared with those in the model control group, but not significantly (P>0.05). The contents of endotoxin (at 6 and 12 h in SAP rats and on 7, 14, 21, and 28 d in OJ rats, respectively) and PLA2 (at 6 and 12 h in SAP rats and on 28 d in OJ rats, respectively) in the treated group were significantly lower than those in the model control group (P<0.01 and P<0.001, respectively). Besides, myocardial pathological injuries were mitigated in SAP and OJ rats.

CONCLUSIONIn this study, we found that Salvia miltiorrhizae improved myocardial pathological changes, reduced the content of PLA2 in blood, and decreased the mortality rates of SAP and OJ rats, exerting protective effects on the hearts of the rats.

Animals ; Endotoxins ; blood ; Heart Injuries ; blood ; drug therapy ; etiology ; pathology ; Jaundice, Obstructive ; blood ; complications ; drug therapy ; Male ; Microscopy, Electron ; Pancreatitis ; blood ; complications ; drug therapy ; Phospholipases A2 ; metabolism ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Salvia ; chemistry ; Survival Rate

BACKGROUND/AIMS: The major compounds of Cochinchina momordica seed extract (SK-MS10) include momordica saponins. We report that the gastroprotective effect of SK-MS10 in an ethanol-induced gastric damage rat model is mediated by suppressing proinflammatory cytokines and downregulating cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase (5-LOX), and the activation of calcitonin gene-related peptide. In this study, we evaluated the gastroprotective effects of SK-MS10 in the nonsteroidal anti-inflammatory drug (NSAID)-induced gastric damage rat model. METHODS: The pretreatment effect of SK-MS10 was evaluated in the NSAID-induced gastric damage rat model using aspirin, indomethacin, and diclofenac in 7-week-old rats. Gastric damage was evaluated based on the gross ulcer index by gastroenterologists, and the damage area (%) was measured using the MetaMorph 7.0 video image analysis system. Myeloperoxidase (MPO) was measured by enzyme-linked immunosorbent assay, and Western blotting was used to analyze the levels of cyclooxygenase (COX)-1, COX-2, cPLA2, and 5-LOX. RESULTS: All NSAIDs induced gastric damage based on the gross ulcer index and damage area (p<0.05). Gastric damage was significantly attenuated by SK-MS10 pretreatment compared with NSAID treatment alone (p<0.05). The SK-MS10 pretreatment group exhibited lower MPO levels than the diclofenac group. The expression of cPLA2 and 5-LOX was decreased by SK-MS10 pretreatment in each of the three NSAID treatment groups. CONCLUSIONS: SK-MS10 exhibited a gastroprotective effect against NSAID-induced acute gastric damage in rats. However, its protective mechanism may be different across the three types of NSAID-induced gastric damage models in rats.
Animals ; Anti-Inflammatory Agents, Non-Steroidal/adverse effects ; Arachidonate 5-Lipoxygenase/drug effects ; Calcitonin Gene-Related Peptide/drug effects ; Cyclooxygenase 1/drug effects ; Cyclooxygenase 2/drug effects ; Disease Models, Animal ; Gastric Mucosa/chemistry/drug effects ; Group IV Phospholipases A2/drug effects ; Male ; Momordica/*chemistry ; Peroxidase/drug effects ; Plant Extracts/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Seeds/*chemistry ; Stomach Ulcer/chemically induced/*prevention & control ; Treatment Outcome

OBJECTIVETo explore the role of massive escharectomy at early postburn stage in the prevention of internal organ dysfunction.

METHODS(1) Ten cases of severely burned patients were randomly divided into early (A) and non-early escharectomy (B) groups in equal number. Venous blood samples were harvested from the patients of the two groups in 1, 3 and 7 postburn days (PBDs), And the samples from 6 healthy volunteers were taken as the control. The serum was separated from the above blood samples and was employed to stimulate cultured HUVECs in vitro. The cell viability and permeability was observed after the stimulation. (2) Seventy Wistar rats inflicted with 30% TBSA III degree scalding were used as an animal model, and were randomized into early (C, n = 30) and non-early escharectomy (D, n = 30) groups, with 5 normal rats as control in each group. Intra-peritoneal fluid infusion was carried out at 1, 3, 6, 12, 24 and 48 postburn hours (PBHs) in rats in both groups. The rats were killed by blood letting at 1 hour after fluid supplementation. The changes in peritoneal macrophage (M Phi) activation state and plasma contents of LPS, IL-8, PLA(2) and MDA were determined at 48 hours after escharectomy in the rats.

RESULTSThe cell viability and permeability of the HUVECs co-cultured with the serum from burn patients in E group was much better preserved than that in B group. On the other hand, the peritoneal M Phi activation and the plasma contents of LPS, IL-8, PLA(2) and MDA in C group were obviously decreased compared with those in D group.

CONCLUSIONEarly postburn escharectomy to remove denatured burned tissue were proved to be helpful in ameliorating endothelial injury and in inhibiting activation of inflammatory cells. Therefore, early escharectomy was assumed to be beneficial in the prevention of postburn SIRS and MODS.

Adult ; Animals ; Burns ; blood ; complications ; surgery ; Cell Division ; Cell Line ; Cell Membrane Permeability ; Culture Media, Conditioned ; chemistry ; Dinoprostone ; metabolism ; Endothelium, Vascular ; cytology ; Female ; Humans ; Interleukin-8 ; blood ; Lipopolysaccharides ; blood ; Macrophages, Peritoneal ; metabolism ; Male ; Malondialdehyde ; blood ; Multiple Organ Failure ; etiology ; prevention & control ; Nitric Oxide ; metabolism ; Phospholipases A ; blood ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Time Factors ; Tumor Necrosis Factor-alpha ; metabolism