Objective To investigate the effects of diazoxide (DIA)cardioplegic solution on the reduction of donor cardiomyocyte apoptosis, Methods In a Krebs-Henseleit (KH) solution perfused isolated rabbit heart Langendorff model, 32 rabbit hearts were divided into four groups with simple random sampling (8 rabbits in each group ): DIA group (50μmol/L diazoxide mixed in KH solution),STH group (ST, Thomas' solution), 5-HD group (50μmol/L diazoxide and 100μmol/L 5-hydroxydecanoic acid mixed in KH solution), KH group (KH solution), The rabbit hearts of each group underwent 6 hours of hypothermic (4 C) storage in the corresponding cardioplegic solution. Left ventricular developed pressure (LVDP), maximal values of positive rate of left ventricular pressure (+dp/dtmax) were measured before and after storage, The post-storage values of LVDP and +dp/dtmax were expressed as the percentage of pre-storage control values. Apoptotic cardiomyocytes were detected by the TdT- mediated dUTP-biotin nick end labeling (TUNEL). Malonaldehyde (MDA) contents and adenosine triphosphate (ATP) contents were also measured after storage. Results Recovery rates of LVDP, +dp/dtmax, and ATP contents in DIA group were higher than those of other 3 groups respectively(P〈0. 05), Cardiomyocytes apoptosis percentage and MDA content were lower than other 3 groups respectively(P〈0. 05), Conclusions Diazoxide cardioplegic solution can protect the isolated hearts and this may be relates to opening selective mitochondrial KATP channels. The selective mitochondrial KATP channel antagonist 5-hydroxydecanoic acid can block the cardioprotective effect of diazoxide.
Objective To observe the influences of depolarized arrest and hyperpolarized arrest on alternation of fluidity of myocardial cell membrane during cardiopulmonary bypass (CPB) and evaluate the protective effects on myocardium of hyperpolarized arrest. Methods Seventy-two felines were randomized into three groups, each group 24. Control group: 180 minutes of CPB was conducted without aortic and vena caval cross-clamping. Depolarized arrest group: hearts underwent 60 minutes of global ischemia after aortic cross-clamping (ACC) followed by 90 minutes of reperfusion. The cardioplegic solution consisted of St. Thomas solution (K+16mmol/L). Hyperpolarized arrest group: the protocol was the same as that in depolarized arrest group except that the cardioplegic solution consisted of St.Thomas solution with pinacidil (50 mmol/L,K+5mmol/L). Microviscosity, the reciprocal of fluidity of myocardial membrane was measured in all groups by using fluorescence polarization technique. (Results )Microvis cosity of myocardial cell in depolarized arrest group during ACC period was significantly higher than that before ACC and kept on rising during reperfusion period. Microviscosity of myocardial cell in hyperpolarized arrest group during ACC was trending up and reperfusion periods as well, but markedly lower compared to that in depolarized arrest group at corresponding time points(Plt;0.01). Conclusion Hyperpolarized arrest is more effective in protecting myocardial cells from ischemia-reperfusion injury than depolarized arrest during CPB by maintaining better fluidity of myocardial membrane.
Objective To investigate Kir4.1 expressions in Muuml;ller cells under high glucose conditions and treatment of pigment epitheliumderived factor (PEDF). Methods Cultured rat Muuml;ller cells were divided into control group (5 mmol/L glucose), high glucose group (25 mmol/L glucose), PEDF treatment group (25 mmol/L glucose+100 ng/ml PEDF) and intervention control group(25 mmol/L glucose+phosphate buffer solution). Kir4.1 expressions were measured by Western blot and real-time reverse transcription polymerase chain reaction (RT-PCR). Reactive oxygen species (ROS) productions were measured using 2prime;7prime;dichlorofluorescin diacetate and glutathione peroxidase (GPx)expressions were studied by real-time RT-PCR. Results By Western blot and real-time RT-PCR, it was found the expressions of Kir4.1 decreased obviously under high glucose conditions (real-time RT-PCR: t=4.12, P<0.05; Western blot: t=3.53,P<0.05); simultaneously, ROS generation was increased (t=3.76,P<0.05)and GPx level was decreased (t=3.18,P<0.05). PEDF treatment inhibited the high glucose-induced Kir4.1 down regulation (real-time RT-PCR: t=3.66, P<0.05; Western blot: t=6.43,P<0.01) and decreased ROS generations (t=4.11,P<0.05) and increased GPx levels (t=5.12,P<0.01). Conclusions The high glucose can supress Kir4.1 expressions in Muuml;ller cells by oxidative stress, and PEDF can ameliorate these effects.
Objective To observe the effects of high concentr at ion glucose on the calcium-activated potassium channel of rabbits′ retinal Müller cells. Methods The rabbits′retinal Müller cells were cultured in vitro under the condition of high concentration glucose, and identified by immunohistochemical staining and transmission electron microscopy. Patch-clamp technique was used to observe the changes of the calcium-activated potassium channel of retinal Müller cells caused by high concentration glucose at different time.Results High concentration glucose could inhibit the calcium-activated potassium channel of cultured retinal Müller cells in a time-dependent manner. Conclusion High concentration glucose may reduce the biological functions of Müller cells by inhibiting calcium-activated potassium channel. (Chin J Ocul Fundus Dis,2003,19:164-167)
ObjectiveTo make a better understanding of potassium channel-complex autoimmune antibodies associated limbic encephalitis, we studied in details with patients of this autoimmune disease accompanying without tumors. MethodsDiagnosis of 3 patients were confirmed by antibody detection in serum or CSF. All the clinical data, including history, CSF data, cranial MRI, EEG, pelvic ultrasound and treatment strategy, were carefully gathered. Two to eleven months follow-up were carried out. Results3 female adult patients showed common initial manifestation of seizures, and changes of consciousness, mental disorder and cognitive impairment. Hyponatremia was found in one LGI1-Ab+ patient. Cranial MRI showed unilateral or bilateral signal changes with limbic system. Changes of CSF and EEG were nonspecific. All 3 patients became recovery in different levels after two to eleven months. ConclusionsPotassium channel-complex antibodies associated encephalitis may be a common type of limbic encephalitis in adults without tumors. Seizures may be the first sign of the disease. Hyponatremia is one of characteristics of LGI1-Ab+ patient. Patients may have a fairly good short outcome.
ObjectiveTo observe the changes in open probability and protein expression of large conductance Ca2+-activated K+ (BK) channel in retinal vascular smooth muscle cells (RVSMCs) of diabetic rats. MethodsStreptozotocin (STZ)-induced rat diabetic animal model was established by STZ injection intraperitoneally.RVSMCs were isolated by enzyme digestion. The BK currents in control and diabetic groups were recorded by patch clamp technique in single channel configuration. BK channel protein expression in control and diabetic group were measured by Western blot. ResultsCompared with control group, the NP0 of BK channels in diabetic group were significantly increased (t=4.260, P < 0.05). Compared with control group, there was no significant difference inα-subunit protein expression in diabetic group in RVSMCs (t=10.126, P > 0.05); however, β1-subunit protein expression was remarkably increased in diabetic group (t=5.146, P < 0.05). ConclusionThe NP0 of BK channels andβ1-subunit protein expression are increased in RVSMCs of diabetic rats.