OBJECTIVETo investigate the protective effect of luteolin on isolated rat heart in hypothermic preservation.

METHODSForty male SD rats were randomly divided into 4 groups (n = 10): control group, luteolin low-dose group (7.5 micromol/L), middle-dose group (15 micromol/L) and high dose group (30 micromol/L). Langendorff model of isolated rat heart was used. After 30 min basal perfusion, the hearts were stored in University of Wisconsin solution (UW solution) at 4 degrees C with luteolin (7.5, 15 and 30 micromol/L) or without luteolin for 12 h and followed by 60 min reperfusion. The recovery of cardiac contractile and diastolic function, coronary flow (CF), creatine kinase (CK) leakage in the coronary effluent, myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULTSThe results revealed that luteolin improved the recovery of left ventricular peak systolic pressure and +/- dp/dtmax dose-dependently and increased coronary flow. The leakage of creatine kinase in the coronary effluent was significantly reduced in luteolin-added hearts. Impairment of myocardial ultrastructure after 12 h hypothermic preservation was obviously alleviated in hearts luteolin-added group compared with that in control group. There were no differences between the groups in myocardial water contents.

CONCLUSIONLuteolin as a supplementation in cardiac preservation solution can significantly improve the hypothermic preservation effects on rat heart and have myocardial protection effect, especially in luteolin-added with 30 micromol/L.

Animals ; Cryopreservation ; In Vitro Techniques ; Luteolin ; pharmacology ; Male ; Myocardium ; Organ Preservation ; methods ; Organ Preservation Solutions ; Rats

Generally, mammalian cells and living tissues can be cryopreserved in a frozen state at very low temperatures over a long storage term. The survival rate of cell suspensions is often acceptable however, living tissues suffer a variety of injuries. In this paper, it was demonstrated that the addition of polyphenols extracted from green tea to conventional cell culture medium and tissue compatible liquid, can control cell proliferation and also preserve tissues for several months at ordinary room temperature, including such tissues as blood vessels, cartilage, islet cells and corneas. This protocol allows a non-frozen living tissue bank to be established using the preservation fluid described.
Animals ; Flavonoids/*pharmacology ; Humans ; Organ Preservation Solutions/*pharmacology ; Phenols/*pharmacology ; Tea/*chemistry ; *Tissue Banks ; *Tissue Preservation ; *Tissue Transplantation ; Transplantation, Homologous

OBJECTIVETo investigate whether the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during hypothermic preservation of the rat heart.

METHODSThe Langendorff model of isolated rat heart was used. After equilibrium, the hearts were stored in Celsior cardioplegia solution at 4 degree with or without supplement of diazoxide for 3 or 8 h followed by 60 minutes reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULT(1) Treatment of diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes, lactate dehydrogenase (LDH) and creatine kinase (CK), at the 2nd and 4th minute of reperfusion of rat heart after hypothermic preservation for 3 h. (2) After hypothermic preservation for 8 h, diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes (LDH, CK and glutamic oxalic transaminase) during reperfusion. Moreover, left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution. (3) Diazoxide significantly decreased the water content of myocardium and increased coronary flow of the hearts compared with those in control after hypothermic preservation for 8 h. (4) Impairment of myocardial ultrastructure after 8 h hypothermic preservation was alleviated in hearts treated with 30 mol/L diazoxide. (5) The cardiac effects of 30 mol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (100 micromol/L).

CONCLUSIONDiazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during hypothermic heart preservation via opening of mitochondrial K(ATP) channel.

Animals ; Cardioplegic Solutions ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; drug effects ; Rats ; Rats, Sprague-Dawley

Drugs, Chinese Herbal ; pharmacology ; Heart ; Humans ; Liver ; Organ Preservation Solutions ; Panax ; Pancreas ; Salvia miltiorrhiza

OBJECTIVETo observe the effect of Jisheng injection (JSI) in protecting heart.

METHODSIsolated heart of rat was preserved in modified Euro-Collins solution (mEC) containing JSI for 20 hrs, and that preserved in simple mEC was taken as control. Then Langendorff isolated rat heart perfusion was conducted. Forty minutes after perfusion, the cardiac function, coronary flow, myocardial water content were determined, and lactate dehydrogenase (LDH), creatine kinase (CK) activity in perfusate, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content in myocardial tissue and pathologic change in myocardium were also observed.

RESULTSThe cardiac function and coronary flow of isolated heart preserved in JSI containing mEC was significantly better than those in the control (P < 0.01), with the LDH, CK activity and MDA content significantly lower (P < 0.01 and P < 0.05), SOD activity significantly higher (P < 0.05) and pathologic injury milder than those in control, but comparison of cardiac water content between the two groups showed insignificant difference.

CONCLUSIONJSI has good cardiac protective and anti-oxidizing effects.

Animals ; Antioxidants ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Rats ; Rats, Wistar ; Reishi ; chemistry ; Superoxide Dismutase ; metabolism

AIMThe study was designed to examine the effects of cryoprotective media, and glycerolating and thawing procedures on human sperm motility and gel penetrating ability.

METHODFifteen unselected donors provided semen varying in quality that was distributed in a factorial design across three cryoprotectants (glycerol, egg yolk-citrate-glucose-glycerol and egg yolk-tris-glucose-glycerol). Also, glycerol was added at room temperature versus at 4 degrees C. Two thaw temperatures were tested (laboratory air temperature for 10 min versus a 65 degrees C waterbath for 4 seconds). The proportion of total and progressively motile sperm was estimated immediately after thawing and following incubation at 35 degrees C for 2 h. Migration of sperm for 30 min at 37 degrees C through polyacrylamide gel was tested.

RESULTSDonors differed greatly, with post-thaw total motility of sperm ranging from 9 to 44% (P<0.05). Egg yolk-citrate-glucose-glycerol and egg yolk-tris-glucose-glycerol were superior to glycerol alone (post-thaw values of 35, 37 and 21%, respectively, P<0.05). This was due primarily to poor sperm survival when semen was cooled to 4 degrees C without glycerol or egg yolk. The two thaw temperatures gave similar results. Sperm migration tests paralleled the motility results, but were more sensitive in detecting differences.

CONCLUSIONEgg yolk, particularly in a tris-based medium that is widely used in domestic animals, improved the cryopreservation of both good and poor quality human semen.

Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Ejaculation ; Glycerol ; pharmacology ; Humans ; Male ; Organ Preservation Solutions ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; drug effects ; physiology ; Tissue Donors

The effects of oxygen partial pressure on cryopreservation of the cells with organ preservation solution were explored. Hypoxic UW solution was made by purging the UW solution with argon. The pig proximal tubule epithelial cells (LLC-PK1 cells) were cryopreserved in hypoxic UW solution (Ar-UW group) or standard UW solution (UW group) at 4 degrees C for 48 h. Trypan blue staining and LDH detection were performed to evaluate the injury of the cells. The results showed that the oxygen partial pressure in Ar-UW group was significantly declined from 242+/-6 mmHg to 83+/-10 mmHg. After cryopreservation at 4 degrees C for 48 h, LDH leakage rate and Trypan blue-stained rate in Ar-UW group were (11.3+/-3.4)% and (10.5+/-4.7)%, respectively, which were significantly lower than in UW group [(49.5+/-6.9)% and (47.6+/-9.3)% respectively, both P<0.01]. It was concluded that lower oxygen partial pressure of UW solution was more beneficial to the cryopreservation of LLC.
Adenosine ; Allopurinol ; Cell Hypoxia ; Cell Line ; Cryopreservation ; Cryoprotective Agents ; Epithelial Cells/*cytology ; Glutathione ; Insulin ; Kidney Tubules, Proximal/cytology ; Organ Preservation Solutions ; Oxygen/pharmacology ; Raffinose ; Swine ; Tissue Preservation/methods

Prolongation of the duration of heart preservation in vitro is very important in clinical heart transplantation. Previous studies have shown that mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) plays an important role in cardioprotective effect. The purpose of this study was to assess whether the mitoK(ATP) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during long-term hypothermic preservation of the rat heart. Langendorff model of isolated rat heart was used. After 30 min stabilization of perfusion, the hearts were stored in Celsior cardioplegia solution at 4 degrees C with (15, 30 and 45 micromol/L) or without diazoxide, a mitoK(ATP) channel opener, for 10 h followed by 60 min reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed. We found that: (1) Diazoxide treatment improved the recovery of left ventricular developed pressure and +/-dp/dt(max) dose-dependently. Left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution after hypothermic preservation for 10 h. (2) Diazoxide at 30 and 45 micromol/L significantly decreased the water content of myocardium and increased coronary flow of the hearts compared to those in control. (3) The leakage of myocardial enzymes (lactate dehydrogenase, creatine kinase and glutamate-oxaloacetate transaminase) in the coronary effluent was significantly reduced in diazoxide-treated hearts. (4) Impairment of myocardial ultrastructure after 10 h hypothermic preservation was alleviated in hearts treated with 30 micromol/L diazoxide. (5) The cardiac effects of 30 micromol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (5-HD, 100 micromol/L). These results indicate that diazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during long-term hypothermic heart preservation via opening of mitochondrial K(ATP) channel.
Animals ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; In Vitro Techniques ; Male ; Mitochondria, Heart ; metabolism ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

BACKGROUNDThe composition of the lung preservation solution used in lung graft procurement has been considered the key to minimize lung injury during the period of ischemia. Low-potassium dextran glucose (LPDG), an extracellular-type solution, has been adopted by most lung transplantation centers, due to the experimental and clinical evidences that LPDG is superior to intracellular-type solutions. Ulinastatin has been shown to attenuate ischemia-reperfusion (I/R) injury in various organs in animals. We supposed that the addition of ulinastatin to LPDG as a flushing solution, would further ameliorate I/R lung injury than LPDG solution alone.

METHODSTwelve male New Zealand white rabbits were randomly divided into 2 groups. Using an alternative in situ lung I/R model, the left lung in the control group was supplied and preserved with LPDG solution for 120 minutes. In the study group 50,000 U/kg of ulinastatin was added to the LPDG solution for lung preservation. Then re-ventilation and reperfusion of the left lung were performed for 90 minutes. Blood gas analysis (PaO₂, PaCO₂), mean pulmonary artery pressure (MPAP) and serum TNF-α level were measured intermittently. The pulmonary water index (D/W), tissue myeloperoxidase (MPO) activity, tissue malondialdehyde (MDA) content and morphologic changes were analyzed.

RESULTSThe study group showed significantly higher PaO₂ and lower MPAP at the end of reperfusion. Serum TNF-α level, left lung tissue MPO and MDA in the study group were significantly lower than those in the control group. D/W and pathologic evaluation were also remarkably different between the two groups.

CONCLUSIONSThis study indicated that better lung preservation could be achieved with the use of an ulinastatin modified LPDG solution. Ulinastatin further attenuated lung I/R injury, at least partly by reducing oxidative reactions, inhibiting the release of inflammatory factors and neutrophils immigration.

Animals ; Glycoproteins ; pharmacology ; Lung ; drug effects ; metabolism ; Lung Transplantation ; Male ; Organ Preservation Solutions ; chemistry ; pharmacology ; Rabbits ; Random Allocation ; Reperfusion Injury ; prevention & control

OBJECTIVETo evaluate the effects of tannic acid (TA) treatment on the physico-chemical properties of glutaraldehyde (Glut)-fixed bovine jugular vein (BJV).

METHODSFresh BJVs were treated with Glut or Glut/TA, respectively. The shrinkage temperature, resistance to collagenase or elastase digestion, bio-mechanical properties, and molecular structure of these prepared BJVs were evaluated by Fourier transform infrared spectroscopy.

RESULTSTA treatment resulted in higher shrinkage temperature (P < 0.01), higher resistance to collagenase or elastase digestion (P < 0.01), slightly increased tensile strength (P < 0.01), and elongation at break (P < 0.05) in Glut/TA BJV walls when compared with those of Glut group. Chemical bonds existed between TA and JBV tissue.

CONCLUSIONTA treatment can effectively improve the physicochemical properties of Glut-fixed BJVs.

Animals ; Cattle ; Chemical Phenomena ; drug effects ; Elasticity ; drug effects ; Glutaral ; pharmacology ; Jugular Veins ; pathology ; Organ Preservation Solutions ; pharmacology ; Tannins ; pharmacology ; Tensile Strength ; drug effects ; Tissue Fixation ; methods ; Tissue Preservation ; methods