Ardipusilloside-I stimulates gastrointestinal motility and phosphorylation of smooth muscle myosin by myosin light chain kinase

Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of Ca²⁺-calmodulin using the activities of 20 kDa myosin light chain (MLC₂₀) phosphorylation and myosin Mg²⁺-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low Ca²⁺, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high Ca²⁺, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of MLC₂₀ and Mg²⁺-ATPase activities of Ca²⁺- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.

Screening of effective parts in Jiaoai Decoction with blood enriching function in blood deficiency model rats / 中草药

Objective: To screen the active parts with enriching the blood effects of different solvent extraction parts from Jiaoai Decoction (JAD) by comparing the peripheral blood, immune organ index, interleukin 2 (IL-2), and erythropoietin (EPO) and the effect of content of erythrocyte membrane and energy metabolism enzyme activity of blood deficiency model rats. Methods: Taking the fundus venous plexus blood 5 mL/kg every day, continuous bleeding for 12 d, to establish the rat model of blood deficiency; The model animals in each group were ig given JAD and n-butanol, petroleum ether, ethyl acetate, parts of water extract (12 g/kg crude drug) with compound E-Jiao slurry as a positive control. On day 12 of modeling, the erythrocyte count (RBC), hemoglobin (HGB), red blood cell hematocrit (HCT), platelet count (PLT) were detected in rats and immune organ indexes of spleen and thymus were calculated, the levels of IL-2 and EPO content were detected, energy metabolism enzymes (Na+, K+-ATPase and Ca2+, Mg2+-ATPase) activity were investigated; The difference of the chemical constituents in the different solvent extraction of JAD was analyzed by UPLC. Results: Compared with the control group, the RBC, HGB, HCT, PLT, thymus index, and IL-2 levels of rats in the model group decreased significantly, Na+, K+-ATPase and Ca2+, Mg2+-ATPase activity weakened significantly, the spleen index and EPO levels were significantly increased, indicating that blood deficiency model was successfully established. Compared with the model group, HCT, RBC, HGB, and PLT (P<0.05, 0.01) of rats in n-butyl alcohol group could be significantly increased; The spleen index and the content of EPO (P<0.05, 0.01) were reduced; The thymus index and the content of IL-2 (P<0.01) were increased, the erythrocyte Ca2+, Mg2+-ATPase and Na+, K+-ATPase activity (P<0.05, 0.01) was elevated. The contents of PLT and IL-2 (P<0.01) in petroleum ether group could be obviously increased; The content of EPO and the spleen index (P<0.05, 0.01) were decreased. The ethyl acetate fractions could obviously reduce the spleen index and the content of EPO (P<0.01), increase Ca2+, Mg2+-ATPase activity in erythrocytes (P<0.05). In water group PLT (P<0.05) could be significantly increased and the content of EPO (P<0.01) be decreased. There were no coffee acid, benzoyl paeoniflorin, and ammonium glycyrrhizinate in other solvent extracts of JAD, while the content of gallic acid, protocatechuic acid, ligustrazine phosphate, and chlorogenic acid in n-butanol extract were higher than those in ethyl acetate and water extract parts of JAD. Conclusion: The n-butanol extract is the most active part of JAD for enriching the blood.

Contrastive study on the effect of 3, 5-diiodotyrosine and potassium iodide on myocardial ATPase in hyperthyroidism Wistar rats / 中华地方病学杂志

Objective To study the effects of 3,5-diiodotyrosine (DIT) and potassium iodide (KI) on myocardial ATPase activity in hyperthyroidism Wistar rats induced by thyroid tablets.Methods Seventy-two Wistar rats were divided into 8 groups according to body weight by the random number table method (9 rats in each group),respectively,which were control group,hyperthyroidism model group,low,medium and high doses groups (both DIT and KI contents were 25.0,166.7,500.1 μg/kg).Physiological saline was intragastrically administrated to the control group;the hyperthyroidism model group was given thyroid tablet suspension (200.0 mg/kg);DIT and KI groups were given thyroid tablet suspension with corresponding doses of iodine simultaneously.The medicine was given once a day for a mouth,all the rats were sacrificed and heart tissue was collected.The colorimetric method was used to examine the activity of ATPases (Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase).Results The activities of Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase were significantly different statistically between groups (F =2.99,3.03,6.18,all P ＜ 0.01).Compared with the control group [(4.01 ± 0.22),(4.28 ± 0.28),(4.46 ± 0.35) μmol/mg·h],the activities of ATPases (Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase included) were reduced significantly in hyperthyroidism model group [(3.60 ± 0.25),(3.42 ± 0.31),(3.85 ± 0.17)μ mol/mg·h,all P ＜ 0.01];the activities of Mg2+-ATPase in DIT medium dose group [(3.89 ± 0.35)μmol/mg ·h],Ca2+-ATPase in DIT medium and high doses groups [(4.12 ± 0.20),(4.09 ± 0.21)μ mol/mg·h] were reduced significantly (all P ＜ 0.05);the activities of Na+-K+-ATPases,Ca2+-ATPase were decreased significantly in three KI groups [(3.64 ± 0.32),(3.60 ± 0.32),(3.53 ± 0.33),(3.93 ± 0.22),(3.90 ± 0.23),(3.85 ± 0.26)μmol/mg·h],Mg2+-ATPase in KI high dose group [(3.65 ± 0.49)μmol/mg·h] was decreased significantly (P ＜ 0.05or ＜ 0.01).Compared with the hyperthyroidism model group,the activities of ATPase were increased in most of the DIT groups [Mg2+-ATPase in low,medium doses groups:(4.06 ± 0.51),(3.89 ± 0.35)μmol/mg·h;Ca2+-ATPase in low,medium,high doses groups (4.15 ± 0.26),(4.12 ± 0.20),(4.09 ± 0.21)μmol/mg·h,all P ＜ 0.05].Conclusion Supplementation of thyroid tablets in the process of hyperthyroidism formation in Wistar rats will reduce myocardial damage by DTT compared with the same dose of KI.

Effects of percutaneous midband pulse current stimulation in hepatic region on the activity of hepatic mitochondrial Na+-K+-ATPase and Ca2+-Mg2+-ATPase in exercise-induced fatigued rats / 解放军医学杂志

Objective To explore the effects of percutaneous impulsive current stimulation in hepatic region on the activity of hepatic mitochondrial Na+-K+-ATPase and Ca2+-Mg2+-ATPase in exercise-induced fatigued rats, in order to investigate the effect of exercise-induced fatigue. Methods Seventy-two 8-week old male Wistar rats were randomly divided into 4 groups (18 each): control group (group A), fatigue group (group B), stimulation before fatigue group (group C) and stimulation after fatigue group (group D). Exhaustion of animals in B, C and D groups were reproduced by prolonged swimming. Current stimulation (1024Hz, 10mA, current cycle 1sec) for 20 minutes was given to the rats of group C before swimming, and to those in group D after exhaustion. At the weekend of 1st, 3rd and 5th week after modeling, the rats were sacrificed in batches from each group (6 each). The activities of hepatic mitochondrial Na+-K+-ATPase and Ca2+-Mg2+-ATPase were determined by spectrophotometry, and Bradfood protein quantification was employed to quantitate the protein in rats' hepatic mitochondria. Results No significant difference was found in swimming-exhaustion time among 3 groups at the first weekend (P>0.05), while the swimming-exhaustion time was significantly prolonged at the 3rd and 5th weekends in group D than in group B and C (P+-K+-ATPase and Ca2+-Mg2+-ATPase among the 4 groups (including group A) at the weekend of the 1st week (P>0.05), while the enzyme activities were obviously lower at the 3rd and 5th weekend in group B than that in groups A, C and D (P+-K+-ATPase and Ca2+-Mg2+-ATPase. Percutaneous pulsive current stimulating hepatic region of exercise-induced fatigued rats may improve the enzyme activity, reduce the concentration of free calcium and calcium overload in mitochondria, stimulate the oxidative phosphorylation, accelerate the rate of respiratory chain, promote exercise endurance and score, and relieve exercise-induced fatigue rapidly.

Effects of Yiqihuoxuejiedu Formula on the Activity of G-6-Pase and Mg~(2+)-ATPase of Mice with Lewis Lung Cancer / 中国中医药信息杂志

Objective To observe the effect of Yiqihuoxuejiedu formula on the activity of G-6-Pase and Mg2+-ATPase of Lewis cells of mice with Lewis lung cancer. Method Enzyme cytochemistry was used to detect the activity of G-6-Pase and Mg2+-ATPase of the mice lung cancer of Lewis cells. Result The response granules of G-6-Pase and Mg2+-ATPase in the mice lung cancer of Lewis cells became smaller after the treatment of the Yiqihuoxuejiedu formula, the amount of the granules became fewer, the density was lower, which indicated the activity of enzyme reduced evidently. Conclusion Yiqihuoxuejiedu formula can decrease the activity of the G-6-Pase and Mg2+-ATPase.

STUDY ON MECHANISM OF PLATELET ACTIVATION IN VASCULAR DEMENTIA / 解放军医学杂志

To explore the mechanism of platelet activateation in vascular dementia(VD), the activity of myosin light chain phosphorylation and Ca 2+ Mg 2+ ATPase,the concentration of [Ca 2+ ]i were measured in 28 healthy controls, 38 patients with acute cerebral infarction (ACI), and 32 patients with VD. The activity of MLCK in VD and ACI groups was significantly higher than that in the controls( P

Effect of Qinglongyi polysaccharide on sialic acid level, ATPase activity, and membrane potential in erythrocytes of S_(180) mice / 中草药

Objective To study the effects of Qinglongyi polysaccharide on sialic acid (SA) content, Na+, K+-ATP and Ca2+, Mg2+-ATPase activity, and membrane potential in erythrocytes of S180 mice. Methods Using Kit to determine the SA content and Na+, K+-ATPase, Ca2+, Mg2+-ATPase activity, using flow cytometry to determine the membrane potential of erythrocytes of S180 mice. Results In the treated groups with three doses, it showed Qinglongyi polysaccharide could make the SA content increased (P

Dynamic stuey on intracellular free calcium,ATP level and membrane Ca~(2+)-Mg~(2+) ATPase activity of erythrocyte in patients with acute cerebral infarction / 临床神经病学杂志

Objective To explore the dynamic alteration of intracellular free calcium concentration([Ca 2+ ]i),ATP level and membrane Ca 2+ Mg 2+ ATPase activity of erythrocyte in the patients with acute cerebral infarction(CI).Methods we examined [Ca 2+ ]i,ATP level and membrane Ca 2+ Mg 2+ ATPase activity of erythrocyte in 30 patients with acute CI and 28 health controls by Fluorescence Activated Cell Sorter.Results [Ca 2+ ]i in erythrocyte increased significantly in CI group( P

Altered kinetic properties of liver mitochondrial membrane-bound enzyme activities following paracetamol hepatotoxicity in the rat.

The effects of treatment with subtoxic (375 mg/kg) and toxic (750 mg/kg) doses of paracetamol on NADH oxidase, succinoxidase and Mg2+-ATPase activities in rat liver submitochondrial particles were examined. In the NADH oxidase system, treatment with subtoxic doses of paracetamol resulted in a 37% increase in activation energy in the high temperature range (E1) while the phase transition temperature (Tt) for this system decreased by 9°C. Subtoxic doses caused a 43% decrease in E1. For the succinoxidase system, Tt decreased by 2·4 to 3·4°C after paracetamol administration. E2 increased by 42% only in the subtoxic-treatment group while E1 remained unaltered in both paracetamol-treated groups. For the Mg2+-ATPase system, subtoxic doses of paracetamol treatment did not change the values of E1, E2 and Tt whereas toxic dose treatment resulted in a 29% decrease in E2 with a concomitant increase in Tt by 2·4°C without any change in the value of E1 · The results thus suggest that treatment with toxic and subtoxic doses of paracetamol results in possible differential alterations in the membrane lipid milieu.

Activity of Ca~(2+).Mg~(2+)-ATPase and SDH in myocardium of rat induced by macleaya cordata and ultrastructures / 中国法医学杂志

Objective To study the mechanism of the rat’s myocardium induced by macleaya cordata. Methods36 SD rats were randomly assigned to 2 experimental groups treated by macleaya cordata decoction with a dose of 1/6 LD50, 1/3 LD50 respectively and a control group treated with distilled water by oral administration. The rats were killed 6 hours after the administration for taking the cardiac muscle to examine the Ca2+.Mg2+-ATPase, SDH while observing the structures. ResoultsIn the group treated by macleaya cordata with dose of 1/6 LD50, the activity of Ca2+.Mg2+-ATPase and SDH were increased significantly.However, the groups treated by macleaya cordata with dose of 1/3 LD50 and distilled water, the activity of Ca2+.Mg2+-ATPase,SDH showed no significant change. The apoptosise in myocardium cells of rat’s can be observed in the experimental groups,while the changes were not seen in control group. ConclusionThe results indicate that the macleaye cordata can induce and improve the apoptosis of myocardium cells in low dose, and these effects may be induced by stimulating the activity of Ca2+.Mg2+-ATPase,SDH in myocardium cells.

Changes of Activities of Na~+K~+-ATPase,Ca~(2+)Mg~(2+)-ATPase in Erythrocyte Membrane and Blood Viscosity in Children with Essential Hpertension / 实用儿科临床杂志

Objective To study the changes of Na+K+-ATPase,Ca2+Mg2+-ATPase activities in erythrocyte membrane and blood viscosity in children with essential hypertension.Methods The activities of Na+K+-ATPase,Ca2+Mg2+-ATPase in erythrocyte membrane were determined by a colorimetric method.Blood viscosity was measured and analyzed with the statistic analysis SPSS 12.0 software in 50 children from Nov.2004 to Dec.2004 in the people's hospital of guizhou province and adolescents with essential hypertension.Thirty healthy children were collected as control group.Results The activities of Na+ K+-ATPase[(6.12?1.30)?molpi/(gHb?h)and(4.59?1.40)?molpi/(gHb?h)],Ca2+Mg2+-ATPase[(7.46?1.30)?molpi/(gHb?h)and(5.81?1.20)?molpi/(gHb?h)] were lower significantly in hypertension group than those in control group(Pa

EFFECTS OF GINSENOSIDES ON THE ACTIVITIES OF Ca~(2+)-ATPase & Mg~(2+)-ATPase IN RABBIT CEREBRUM IN VITRO / 中国药理学通报

The effects of saponins extracted from stems & leaves of Panax ginseng C.A.Meyer including the total ginsenoside ( GNS) , panaxad-iol saponin ( PDS ) & panaxatriol saponin ( PTS ) on the activities of Ca2+-ATPase & Mg2+-ATPase in rabbit cerebral microsomes were studied in vitro. It was found that GNS & PTS significantly inhibited the activities of Ca2+-ATPase & Mg2+-ATPase. The activity of Ca2+-ATPase was activated by PDS at the concentrations of 10 & 100mg/L, and it dramatically inhibited the activity of Ca2+-ATPase the concentration of 1000mg/L. Furthermore, the activity of Mg2+-ATPase was inhibited by PDS at the concentrations of 100 & 1000mg/L. Chlorpromazine ( 0 .35 & 0 .70 mmol/L ) possessed the inhibitory effects on the activities of both Ca2+-ATPase & Mg2+-ATPase.

CLINICAL STUDY ON Ca~(2+)-Mg~(2+)-ATP ENZYME ACTIVITY IN RED CELL MEMBRANES OF DIABETIC PATIENTS / 中华内分泌代谢杂志

The basal and calmodulin stimulated Ca2+-Mg2+-ATPase activities in erythrocyte membranes were measured in 40 diabetic patients and 36 normal subjects as control. The results showed that the enzyme activity is significantly decreased in diabetics. The reduction of enzyme activity is inversely related to serum glucose as well as glycosylated hemoglobin. It is suggested that the function of Ca2+-Mg2+-ATPase in red cell membrane of diabetics is decreased. This may be one of the factors causing diabetic microangiopathy. Furthermore, the reduction of enzyme activity might casually be related with hyperglycemia.

Protective mechanism of metallothionein on cultivated rat cardiomyocytes in hypoxic preconditioning / 中国病理生理杂志

AIM: Studying the mechanism of protective role of metallothionein (MT) in hypoxic preconditioning(HPC) of cultivated rat cardiomyocytes. METHODS: Using the model of hypoxia/reoxygenation of cultivated rat cardiomyocytes. Determing the contents of MT, malonyldialdehyde (MDA) - metabolism product of lipid peroxidation and the activities of Na+ - K+ ATPase, Ca2+ - Mg2+ ATPase of cardiomyocytes 24 h after HPC, the determining the relevant changes after using MT antibody. RESULTS: After 24 h in HPC, the contents of MT and activities of Na+ - K+ ATPase, Ca2+ - Mg2+ ATPase were obviously higher than those in the control and hypoxia/reoxygenation(P＜ 0. 05 ), and the contents of MDA were decreased remarkedly (P ＜ 0.01 ). Then after using MT antibody, the activities of two enzyme were progressively decreased and the contents of MDA were significanily higher than those in the control and MT antibody - free groups(P ＜ 0.01 ). CONCLUSION: HPC may induce excessive synthesis of MT, and MT can protect myocardial reoxygenation injury by eliminating lipid peroxidation and rising the activities of Na+ - K+ ATPase and Ca2+ - Mg2+ ATPase.

ERYTHROCYTE MEMBRANE CARBOHYDRATES, LIPIDS AND Ca~(2+)-Mg~(2+) ATPase IN DUCHENNE MUSCULAR DYSTROPHY / 第二军医大学学报

The carbohydrate and lipid contents and Ca2+-Mg2+ ATPase activity of erythrocyte membrane were studied in 10 patients with DMD. The results demonstrated that the sialic acid content was markedly decreased, and the neutral sugar content normal. The cholesterol was significantly increased, but the phospholipids unchanged, resulting in a significantly reduced P/C ratio. The activity of Ca2+-Mg2+ ATPase elevated markedly. The study suggests that the erythrocyte membranes with DMD are altered both in composition and function.

In vitro effect of prostaglandins, prostaglandin endoperoxides and thromboxane on rat intestinal alkaline phosphatase and (Ca2+ – Mg2+) adenosine triphosphatase.

The activity of alkaline phosphate and Ca2+–Mg2+ adenosine triphosphatase, two of the enzymes involved in limpid and calcium uptake across the intestinal membrane, were increased in experimental atherosclerosis. Administration of Annapavala sindhooram, an antiatherosclerotic drug, lowers these enzyme levels to near normal values. Prostaglandin E2 stimulated the enzyme activities in vitro, while prostaglandin endoperoxide inhibited the activity. Thromboxane and other prostaglandins had no effect on the enzyme activities. Addition of the antiatherosclerotic drug to the in vitro assay system reversed the effect of both prostaglandin E2 and prostaglandin endoperoxide.