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OBJECTIVE: To observe the effect of electroacupuncture(EA) at "Shenmen"(HT7) and "Sanyinjiao"(SP6)on energy metabolism in paraventricular nucleus (PVN) of hypothalamus in insomnia rats, so as to explore its mechanism underlying improvement of insomnia. METHODS: A total of 66 SD rats (half male and half female) were randomized into 3 groups：normal control, model and EA groups(n＝22 per group). The insomnia model was established by binding the rat for at least 4 h (step increase of 30 min per day), once daily for 15 days. EA (5 Hz /25 Hz, 0.5－1.0 mA) was applied to unilateral HT7 and SP6 for 15 min, once daily for 5 days. The rats' spontaneous activities during day and night were recorded by using the ClockLab Data Collection and Analysis System, and the duration of exhausted swimming was detected by using load-bearing endurance swimming test. The expression of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) of PVN tissue was assayed by Western blot, and the contents of acetyl coenzyme A (Ac-CoA) and Na＋-K＋adenosine triphosphatase (Na＋-K＋-ATPase) in the PVN tissue, and corticosterone (CORT) in plasma were assayed by ELISA. Changes of the ultrastructure of PVN cells were observed by transmission electron microscope. RESULTS: After modeling, the rats' daytime and nocturnal locomotor activities were significantly increased and decreased, respectively (P<0.05), and the duration of exhausted swimming was considerably shortened in the model group compared with that of the normal control group (P<0.05). The expression level of AMPK protein in the PVN was obviously up-regulated (P<0.05), and the contents of Ac-CoA and Na＋-K＋-ATPase in PVN and CORT in plasma were markedly decreased in the model control group relevant to the normal group (P<0.05). After EA intervention, the increased daytime locomotion and the decreased nocturnal activities, the shortened duration of exhausted swimming, the up-regulated expression of AMPK, and the decreased Ac-CoA, Na＋-K＋-ATPase and CORT contents were all reversed in the EA treated rats relevant to those of the insomnia rats (all P<0.05). Moreover, ultrastructural observation showed mitochondrial swelling and disappearance of partial ribosomes in the plasma of PVN cells in the model group, while in the EA group, only mild swelling of some mitochondria was found, being with basically normal nuclear membrane, mitochondria, rough endoplasmic reticulum, Golgi complex and ribosomes. CONCLUSION: EA at HT7 and SP6 has a positive effect in improving insomnia and insomnia-induced fatigue in insomnia rats, which may be associated with its effects in restraining the expression of AMPK protein, and up-regulating the contents of Ac-CoA and Na＋-K＋-ATPase in PVN and CORT in plasma.
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Objective@#To explore the effects of change of activity of vacuolar adenosine triphosphatase (V-ATPase) of myocardial lysosome on myocardial damage in rats after severe burn and its mechanism.@*Methods@#The myocardial lysosomes were extracted from the hearts of 12 SD rats with ultra-high speed gradient density centrifugation, then Western blotting and transmission electron microscope observation were conducted for identification. One hundred and twenty rats were divided into pure burn group, ATP group, normal control group, and bafilomycin group according to the random number table, with 30 rats in each group. Rats in pure burn group and ATP group were inflicted with 40% TBSA full-thickness scald on the back. Immediately after injury, rats in pure burn group were intraperitoneally injected with lactated Ringer′s solution in 4 mL·%TBSA-1·kg-1, and rats in ATP group were intraperitoneally injected with ATP in 0.4 mg/kg at 12 h before burn, immediately after burn, and 12 h after burn. Rats in normal control group did not receive any treatment, and rats in bafilomycin group were intraperitoneally injected with bafilomycin A1 in 0.3 mg/kg at the same time points as those of ATP group. At 24 h after burn, 30 rats from each group were collected for determining activity of V-ATPase of myocardial lysosome with coupled-enzyme assay and the expression of myocardium autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) and P62 by Western blotting. Left ventricular arterial blood was collected to detect the content of 5 items of myocardial enzyme spectrum and cardiac troponin T (cTnT). Data were processed with one-way analysis of variance and t test.@*Results@#(1) After identification, both the expression level of lysosome-related membrane protein 1 and purity of lysosome in the sample were high, and the structure of lysosome was intact. (2) At 24 h after burn, the activity values of V-ATPase of myocardial lysosome in rats of pure burn group, ATP group, normal control group, and bafilomycin group were (2.03±0.67), (3.01±0.58), (4.29±0.26), and (1.83±0.52) μmol·mg-1·h-1, respectively. The activity value of V-ATPase of myocardial lysosome in rats of pure burn group was significantly lower than the values in ATP group and normal control group (with t values respectively 3.14 and 8.87, P values below 0.01). The activity values of V-ATPase of rats in normal control group were significantly higher than those in bafilomycin group (t=11.87, P<0.01). At 24 h after burn, the expressions of myocardial LC3 and P62 in pure burn group were significantly higher than those in ATP group and normal control group (with t values from 3.73 to 5.88, P values below 0.01). The expressions of myocardial LC3 and P62 in normal control group were significantly lower than those in bafilomycin group (with t values respectively 2.64 and 3.07, P<0.05 or P<0.01). At 24 h after burn, the content of 5 items of myocardial enzyme spectrum and cTnT in pure burn group was significantly higher than that in ATP group and normal control group (with t values from 3.24 to 16.72, P values below 0.01). The content of 5 items of myocardial enzyme spectrum and cTnT in normal control group was significantly lower than that in bafilomycin group (with t values from 2.39 to 10. 70, P values below 0.01).@*Conclusions@#The activity of V-ATPase of myocardial lysosome decreased in rats after severe burn, which can result in myocardial damage by inhibiting myocardial autophagy flux.
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Exosomes are small intracellular membrane-based vesicles with different compositions that are involved in several biological and pathological processes. The exploitation of exosomes as drug delivery vehicles offers important advantages compared to other nanoparticulate drug delivery systems such as liposomes and polymeric nanoparticles; exosomes are non-immunogenic in nature due to similar composition as body׳s own cells. In this article, the origin and structure of exosomes as well as their biological functions are outlined. We will then focus on specific applications of exosomes as drug delivery systems in pharmaceutical drug development. An overview of the advantages and challenges faced when using exosomes as a pharmaceutical drug delivery vehicles will also be discussed.
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Objective To know the effect of nanometer titanium dioxide (nano -TiO2 )on the oxidative stress in zebrafish. Methods A total of 64 zebrafishes were randomly divided into 4 groups with different concentrations of nano -TiO2 exposure,which including control group,low dose group (2 mg/L),moderate dose group (8 mg/L)and high dose group (32 mg/L).The content of MDA,the activities of SOD and ATPase in gill,liver and muscle were measured in 1 0 zebrafishes in each group after 30 d exposure.Histopathological changes of gill,liver and muscle in the remainder fishes were observed.Results The content of Malondialdehyde (MDA)in the fish gill of the moderate and high dose group were significantly higher than that of the control group (P <0.05).The activity of Superoxide dismutase (SOD)in the fish gill of the high dose group was significantly increased compared with the control group (P <0.05).The activities of Adenosine triphosphatase(ATPase)in the fish gill of the moderate and high dose group were significantly decreased compared with the control group (P <0.05).The changes on the content of MDA and the activities of SOD in the fish liver were similar with that in the fish gill after nano -TiO2 exposure,while there was no significant change in the activity of ATPase of the fish liver.And no significant changes were observed in the content of MDA,the activities of SOD and ATPase in the fish muscle (P >0.05 ). Histopathological observation showed infiltration by inflammatory cells, swelling and degeneration, vacuolation and necrosis in zebrafish gill and liver with different degrees.Conclusion Nano -TiO2 exposure could induce the oxidative stress in zebrafish,causing pathological changes in the fish gill and liver.
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Objective: To investigate the expression of copper-transporting P-type adenosine triphosphatase (ATP 7B) in the ovarian cancer and evaluate its relationship with chemo-resistance and the clinicopathologic parameters of ovarian carcinoma. Methods: Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of ATP 7B in 36 cases of ovarian carcinoma, 10 cases of benign ovarian tumor, and 10 cases of normal ovarian tissue. Results: The positive expression rate of ATP 7B in ovarian carcinoma was 38.9% significantly higher than that in benign ovarian tumor (0%) and normal ovarian tissue (0%) (P < 0.01). There was a significant correlation between ATP 7B expression and differentiation degree. ATP 7B expression was significantly higher in poorly differentiated carcinoma than that in well and moderate differentiated carcinoma (P < 0.05). The expression of ATP 7B was more positive in patients who received chemotherapy before surgery than those who did not receive chemotherapy. The difference was significant (P < 0.05). The response rate of ATP 7B-positive patients to chemotherapy was significantly lower than the ATP 7B-negative patients (28.6% vs 72.7%, P < 0.05). Conclusion: These data suggested that ATP 7B plays an important role in the multidrug resistance of the ovarian carcinoma to Pt-based chemotherapy.
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AIM: To study the protective effects of baicalin on myocardial ischemia-reperfusion injury in rats. METHODS: The models of myocardial ischemia-reperfusion injury were established by occluding left anterior descending coronary artery (LAD) for 30 min, followed by reperfusion for 120 min. The rate of rise and decline of left ventricular pressure (±dp/dtmax) and end-diastolic pressure of left ventricle (LVEDP) were monitored continuously with polygraph. After reperfusion, the blood and myocardium samples were taken for determination of malondialdehyde (MDA) content, superoxide dismutase (SOD), Na+-K+-ATPase, Ca2+-ATPase activities in myocardium, creatine kinase (CK) and lactate dehydrogenase (LDH) in serum with spectrophotometer. The ultrastructural changes in ischemic myocardium were assessed by transmission electron microscope. RESULTS:dtmax and LVEDP, decreased plasma CK and LDH levels, reduced myocardial MDA content, and increased the activities of SOD, Na+-K+-ATPase, and Ca2+-ATPase in myocardium following ischemia-reperfusion. The ultrastructural injury in reperfused myocardium was relieved. CONCLUSION: Baicalin possesses a protective effect against myocardial ischnemia-reperfusion injury through scavenging oxide radicals and improving Na+-K+-ATPase and Ca2+-ATPase activities.
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Objective To explore the protective effect of vitamin E on the rabbit bladder after partial outlet obstruction artificially setup. Methods A total of 28 New Zealand white male rabbits were divided into 4 groups (group A in 6,group B in 6,group C in 8 and group D in 8).Group A,B and C were fed a regular diet,and group D were placed on a diet enriched with 600 mg vitamin E.After 4 weeks partial outlet obstruction was created in groups C and D,while group B underwent sham operation. After 4 weeks of obstruction each rabbit was sedated and cystometry was repeated.After cystometry the bladder was weighed.The gene expression of sarcoplasmic endoplasmic reticulum,calcium,magnesium,adenosine triphosphatase (SERCA2) in bladder was detected by using RT-PCR assay,while the protein level of SERCA2 was measured by Western blot analysis. Results All parameters measured were approximately identical in nomal rabbits(Group A) and shum operation rabbits(group B).Thus,these 2 groups were combined as the control group(Group A and B).Partial outlet obstruction resulted in bladder weight increased significantly in obstructed groups given vehicle group C(13.07?1.71)g and those vitamin E group D(11.80?2.01)g,4-fold higher than in the control group (2.81?0.30)g(P
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Aim To research the changes of myocardial endoxin level in rats with myocardial ischemia reperfusion (MIR) and the protevtive effects of anti digoxin antiserum, an endoxin specific antagonist, on MIR injury. Methods Myocardial ischemia reperfusion injury models were obtained by ligating left anterior descending coronary artery 30 min followed by 45 min reperfusion. Sprauge Dawley rats were randomly divided into seven groups each with 10 rats. There were sham group, MIR group, normal saline group, verapamil group, low dose anti digoxin antiserum group, middle dose anti digoxin antiserum group, and high dose anti digoxin antiserum group. After reperfusion of left ventricular myocardium, sample of ischemia were processed immediately. Myocardial endoxin levels, Na +, K + ATPase activities, and intramitochondrial Ca 2+ contents were measured. The myocardial morphology were observed. Results Myocardial endoxin levels were significantly increased; Na +, K + ATPase activities were remarkably decreased; intramitochondrial Ca 2+ contents were remarkably raised. Meanwhile, myocardial morphology injury were remarkable in light microscope and electric microscope. Middle and high dose of anti digoxin antiserum intervention, myocardial endoxin levels were remarkably decreased; Na +, K + ATPase activities were drastically increased; intramitochondrial Ca 2+ declined. The myocardial histological morphology was significantly improved. Conclusion Antidigoxin antiserum, an endoxin antagonist, had protective effect against MIR. The mechanism maybe related to antagonizing endoxin, restoring energy metabolism, attenuating intracellular Ca 2+ overload.
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AIM To evaluate the changes of serum and brain tissue endoxin in model of bilateral cerebral hemisphere ischemic reperfusion injury, and effect of anti digoxin antiserum (an antagonist of endoxin). METHODS The bilateral cerebral hemisphere ischemic model was prepared by ligating three vascular by Kameyama's manner. SD rats were randomly divided into 7 groups and each group had 8 rats. Sham group, ischemic reperfusion group, negative control group, nimodipine group, low concentration anti digoxin antiserum group, middle concentration anti digoxin antiserum group, high concentration anti digoxin antiserum group. The blood was collected at the end of reperfusion, meanwhile rats were killed, and the bilateral cerebral hemisphere were took out and used to prepare encephlon homogenate and made into samples of light microscope. RESULTS Compared with sham group, the serum CK content increased; Brain tissue SOD activity reduced and MDA content increased importantly in ischemia reperfusion group; The levels of serum and brain tissue endoxin in ischemia reperfusion group were significantly higher, while ATPase activity in brain tissue decreased; Mitochondrial Ca 2+ content in brain tissue increased significantly and Mg 2+ content decreased significantly. In brain tissue,there was some inflammatory change and local necrosis;The rank order and structure of cell wasn't clear;The morphology of pyramidal cell was abnormal. Compared with ischemic reperfusion group, Anti digoxin antiserum reduced serum CK content; It antagonized lowering of SOD activity and increase of MDA content in brain tissue; It remarkably reduced the level of brain tissue endoxin; It reduced abnormal ion content of brain tissue mitochondrion induced by cerebral ischemic reperfusion injury; The high and middle concentration anti digoxin antiserum had a significant effect on raising brain tissue ATPase activity. It reduced neuron denaturation. CONCLUSION Cerebral ischemic reperfusion can increase the level of brain tissue and serum endoxin and higher endoxin can promote brain injury. Endoxin is a major factor involved in cerebral ischemic reperfusion injury. Anti digoxin antiserum can reduce brain tissue injury and had a protective and treatment effect on cerebral ischemic reperfusion injury by antagonizing the effect of endoxin.
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OBJECTIVE:To observe the effect of ligustrazin on the mitochondrial damage of myocardium in ischemia-reperfusion rats and the mechanism involved METHODS:Rat model of myocardial ischemia-reperfusion injury was established by coronary artery ligation and the activities of Ca2+-ATPase,Ca2+?Mg2+-ATPase,succinic dehydrogenase(SDH),cytochrome oxidase(CCO),SOD,GSH-PX and contents of phospholipid(PL),MDA,Cyt aa3 and Cyt C in the myocardial mitochondria were observed RESULTS:As compared with ischemia-reperfusion group(IR),IR+L(ligustrazin)group showed significant increase in the activities of Ca2+-ATPase,Ca2+?Mg2+-ATPase,SDH,CCO,SOD and GSH-PX(P
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AIM: To study the mechanism of brain ischemia-reperfusion injury from ATPase activity and free radical metabolism in aged rats. METHODS: The young rats (5 months) and the aged rats (more than 20 months) were divided into young control group(YCG), young model group(YMG), aged control group(ACG) and aged model group(AMG). The ATPase and SOD activities and the contents of MDA, Ca 2+ , Na + and K + were measured in the rats with 30 min brain ischemia followed by 60 min reperfusion. RESULTS: The Ca 2+ content in the AMG was higher than that in the YMG and the ACG. The Na +-K +-ATPase activity in the ACG was lower than that in the YCG,was lower in the AMG than that in the YMG . The Ca 2+ -ATPase activities in the YCG was higher than that in the ACG, was lower in the AMG than that in the YMG and was higher than the ACG's. The serum and brain tissue SOD activities in the ACG was lower than that in the YCG, was lower in the AMG than YMG 's. The serum and brain tissue MDA/SOD ratio in the AMG was higher than that in the ACG. CONCLUSION: The brain tissue ischemia- reperfusion injury was related with calcium overload and free radical injury. The pathological changes were obvious and had some characteristics in the aged rats compared with the young rats because of the brain tissue aging changes in ATPase,calcium content and free radical metabolism in the aged rats.
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To explore the mechanism of protecting erythrocyte deformability (ED) of polysaccharide sulfate(PSS)and propyllene glycolmannurate sulfate(PGMS) in patients with acute myocardial infarction (AMI). The erythrocyte filtration index(EFI),erythrocgte membrane Na+,K+-ATPase .glutathione peroxidase(GSH-Px)activity and lipid peroxide(LPO) were measured in 52 patients with AMI. Meanwhile the effects of PSS and PGMS on EFI, Na+,K+-ATPase ,GSH-Px and LPO in AMI patients were observed in vitro. The results showed that the EFI and LPO were markedly higher ,Na+ ,K+-ATP ase and GSH-Px were si-hnificantly lower in patients than those in control(P0. 05). These results indicate that PSS and PGMS could improve the ED in patients ,and the efficacy of PSS or PGMS was related to enchancement of erythrocyte membrane ATP-ase anci GSH-Px activity.
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Objective: To observe the protective effect of ligustrazine on myocardial ischemia rat induced by isoproterenol. Methods : The model of myocardial ischemia was induced by subtcuaneous injection of isoproterenol (ISP, 5mg?kg -1 ) into rats and repeated the same dose 24 hours later. The activities of Ca 2+ -ATPase、Ca 2+ -Mg 2+ -ATPase and the contents of Ca 2+ in the myocardial mitochondria were observed, respectively. The changes of protein expression of bcl-2 and bax genes were detected by immunohistochemical staining. Results : In the ligustrazine treated group, as compared with those of the model, the activities of Ca 2+ -ATPase、Ca 2+ -Mg 2+ -ATPase in the myocardial mitochondria increased significantly( P 0.05). Conclusion : Ligustrazine possesses protective effects against myocardial ischemia injury via increasing the activities of Ca 2+ -ATPase、Ca 2+ -Mg 2+ -ATPase in the myocardial mitochondria and influencing the expression of bcl-2.
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AIM: To evaluate the antagonistic effect of anti - digoxin antiserum on hypoxic myocardium and its mechanism. METHODS: It was observed that different concentration of anti-digoxin antiserum effect on endoxin and cell membrane ATPase activity in hypoxic myocardium model. RESULTS: The endoxin level was much higher, cell membrane ATPase activity was much lower in hypoxic myocedium than those of noed; anti-digoxin antiserum can resume membrane ATPase activity. CONCLUSION: Rise of endoxin was basic in molecular biology of myocar- dial damage during myocardial hypoxia. Anti - digoxin antiserum decreased myocardial damge and has protective ef- fect on hypoxic myocardium by antagonistic effeCt of endoxin.
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Animal model of aged rat nephrotoxicity was induced by i. p. administrationof gentamycin in a dose of 140mg/kg/day. Part of those rats were treated with CordycepsSinensis(CS), Epimedium(Ep) and Astragalus Membranaccus (AM) in form of decoc-tion per Os and others seryed as control. The results were summarized as. 1. The nephro-toxicity of gentamycin was aggrevated with age. CS, Ep and AM are effective drugs inpreventing the tdeular damage caused by gentamycin in rats. The pathological changes ofrenal tuoules of the rat groups which treated with CS, Ep and AM were less severe thanthat of the control. 2. CS, Ep and AM could prevent the decline of renal cortical Na~+-K~+-ATPase activity of aged rat induced by gentamycin.
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By using Person's separating blood ghost method basically, Na~+-K~+ ATP ase and Ca~(2+)-Mg~(2+) ATPase activties of human erythrocyte membrane were studied. The comparison was made between 11 G6PD deficient subjects and 11 healthy control persons. The results showed that both enzyme activities as well as ouabain inhibition rate were decreased in G6PD deficient individuals (P
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A rat model of renal ischemia (60 min) and reperfusion (30 min) was reproduced in our present studies. The lipid peroxide content, Na and K concentrations in the renal tissue homogenates were measured by the method of thiobarbituric acid reaction and by flamephotometer, respectively. The results showed that in the cortex of the ischemic kidney, the lipid peroxide content decreased by 37.89?11.92% (P
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The effects of ginsenosides(including the total ginsenoside extracted from stems and leaves-GNS, ginsenoside extracted from rots-GRS, panaxadiol saPonin-PDS and panaxatriol saponin-PTS) on the activities of Na+, K+-ATPase of dog heart sarcolemma were studied in vivo and in vitro.Both GNS and GRS showed dramatic inhibitory effects on the activities of Na+, K+-ATPase at 10, 20 and 40 mg/kg .The activities of Na+, K+-ATPase were also significantly inhibited by PDS and PTS at 5, 10 and 20 mg/kg. In vitro, all the ginsenosides used in this experiment, at the concentrations of 0.01, 0.10 and 1.00 g/L, Possessed significant inhibitory effects on the activities of dog heart sarcolemmal Na+, K+-ATPase .These results indicated that the enhancement of the contractility of cardiac muscle by ginsenosides is associated with its inhibition on the activities of Na+, K+-ATPase .
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0.05) was found between the dechlorinated water group and the extract of A.philoxeroides group in the activities of LDH and SDH after 12 h or 20 h of exposure.Conclusion The extract of A.philoxeroides rapidly inhibits ChE,and then the activity of Mg2+-ATPase,which may suppress the release and utilization of ATP in the Oncomelania snails and finally causes death of snails.
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We observed the effects of testosterone (T) on myosin ATPase activity and its isoenzyme distribution of noninfarcted myocardium in male infarcted rats. 21 days after myocardial infarction (MI), both myocardial fiber diameter and myosin ATPase activity were increased in T treated MI group when compared with that of the MI group (P