Evaluation of the antithrombotic activity of Zhi-Xiong Capsules, a Traditional Chinese Medicinal formula, via the pathway of anti-coagulation, anti-platelet activation and anti-fibrinolysis.

Zhi-Xiong Capsules (ZXC) involving Hirudo, Ligusticum chuanxiong, Salvia miltiorrhiza, Leonurus artemisia, and Pueraria lobata, is an empirical prescription used in Chinese clinics applied for treating cerebral arteriosclerosis and blood-stasis in clinic. However, the mechanism of its antithrombotic activity has not been investigated until now. The present study was designed to investigate its antithrombotic effects, the mechanism of ZXC on anti-thrombus action and to identify the main chemical composition of ZXC using HPLC-DAD-ESI-IT-TOF-MS. Two animal models were used to evaluate the antithrombotic effect of ZXC, the arterial thrombosis model and a venous thrombosis model. ZXC prolonged the plasma recalcification time (PRT), the activated partial thromboplastin time (APTT), the thrombin time (TT) and the prothrombin time (PT) and clearly reduced the content of fibrinogen (FIB) obviously in the arterial thrombosis model. Furthermore, it markedly suppressed the level of TXB2 and up-regulated the level of 6-keto-PGF1a. In addition, it significantly up-regulated the level of t-PA and down-regulated the level of PAI-1 (p < 0.05). These results revealed that ZXC played a vital role in the prevention of thrombosis through interacting with multiple targets, including inhibition of coagulation and platelet aggregation and increasing thrombolysis. A total of 23 compounds were identified as the main components of ZXC by HPLC-DAD-ESI-IT TOF-MS.

Virgin olive oil, palm olein and coconut oil diets do not raise cell adhesion molecules and thrombogenicity indices in healthy Malaysian adults.

BACKGROUND/OBJECTIVES: Effects of high-protein diets that are rich in saturated fats on cell adhesion molecules, thrombogenicity and other nonlipid markers of atherosclerosis in humans have not been firmly established. We aim to investigate the effects of high-protein Malaysian diets prepared separately with virgin olive oil (OO), palm olein (PO) and coconut oil (CO) on cell adhesion molecules, lipid inflammatory mediators and thromobogenicity indices in healthy adults. METHODS: A randomized cross-over intervention with three dietary sequences, using virgin OO, PO and CO as test fats, was carried out for 5 weeks on each group consisting of 45 men and women. These test fats were incorporated separately at two-thirds of 30% fat calories into high-protein Malaysian diets. RESULTS: For fasting and nonfasting blood samples, no significant differences were observed on the effects of the three test-fat diets on thrombaxane B2 (TXB2), TXB2/PGF1α ratios and soluble intracellular and vascular cell adhesion molecules. The OO diet induced significantly lower (P<0.05) plasma leukotriene B4 (LTB4) compared with the other two test diets, whereas PGF1α concentrations were significantly higher (P<0.05) at the end of the PO diet compared with the OO diet. CONCLUSION: Diets rich in saturated fatty acids from either PO or CO and high in monounsaturated oleic acid from virgin OO do not alter the thrombogenicity indices-cellular adhesion molecules, thromboxane B2 (TXB2) and TXB2/prostacyclin (PGF1α) ratios. However, the OO diet lowered plasma proinflammatory LTB4, whereas the PO diet raised the antiaggregatory plasma PGF1α in healthy Malaysian adults. This trial was registered at clinicaltrials.gov as NCT 00941837.

The effect of parenteral lipid emulsions on pulmonary hemodynamics and eicosanoid metabolites in preterm infants: a pilot study.

BACKGROUND: Soy-based intravenous fat emulsion (IVFE) is known to cause a rise in pulmonary artery pressure in the preterm infant, thought to be mediated through eicosanoid metabolites of linoleic acid. We compared the effect of soy-based IVFE and an olive-oil-based IVFE containing less than half the content of linoleic acid on pulmonary artery pressure and eicosanoid metabolites in preterm infants receiving parenteral nutrition. METHODS: In this pilot study at a regional neonatal intensive care unit (ICU), infants received either a soy-based or olive-oil-based IVFE as part of an otherwise identical feeding protocol. Pulmonary artery pressure and urinary thromboxane B2 and prostaglandin F1 alpha were measured at baseline and maximum lipid infusion. RESULTS: There was a greater fall in pulmonary artery pressure in the olive-oil-based IVFE group compared with the soy-based IVFE group. A decrease in urine thromboxane/prostaglandin F1 alpha ratio was seen only in the olive-oil-based IVFE group. CONCLUSIONS: In the parenterally fed preterm infant, an olive-oil-based IVFE may have a beneficial effect on pulmonary artery pressure when compared with soy-based IVFE. Effects on pulmonary vascular tone are likely to be mediated through alterations in eicosanoid metabolism. A randomized trial is warranted to compare the effects of different lipid emulsions.

Influence of ShuJinHuoXue tablets on ischemia reperfusion injury of animals' skeletal muscle.

Ischemia-reperfusion (IR) can lead to serious tissue oxidative injury in animals. ShuJinHuoXue tablet (SJHXT) is a Chinese Traditional Medicine which can relax the muscles and stimulate the blood circulation and has been used as a clinical medicine. In the present study, we investigated the effects of SJHXT pretreatment on oxidative injury using an animal model of acute limb IR. Results showed that SJHXT pre-treatment (200, 300 and 400 mg/kg/day) markedly reduced serum endothelin-1 (ET-1), thromboxane B2 (TXB2) levels and thromboxane B2/6-keto- prostaglandin F1α (TXB2/6-Keto-PGF(1α)), wet weight/dried weight (W/D) ratio, myeloperoxidase (MPO), creatine kinase (CK), lactate dehydrogenase (LDH) activities, and increased serum nitric oxide (NO), 6-Keto-PGF(1α) levels and NO/ET-1 ratio in the IR+SJHXT groups. In addition, the SJHXT pre-treatment (200, 300 and 400 mg/kg/day) markedly reduced skeletal muscle Ca²âº, malondialdehyde (MDA) levels, increased Na⁺-K⁺-ATPase, Ca²âº-Mg²âº-ATPase, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities. Our results suggest that SJHXT pre-treatment may improve skeletal muscle blood vessel microcirculation, decrease skeletal muscle oxidative injury and enhance antioxidant enzymes activities in IR animals.

Dietary palmitic and oleic acids exert similar effects on serum cholesterol and lipoprotein profiles in normocholesterolemic men and women.

To compare the effects of dietary palmitic acid (16:0) vs oleic acid (18:1) on serum lipids, lipoproteins, and plasma eicosanoids, 33 normocholesterolemic subjects (20 males, 13 females; ages 22-41 years) were challenged with a coconut oil-rich diet for 4 weeks. Subsequently they were assigned to either a palm olein-rich or olive oil-rich diet followed by a dietary crossover during two consecutive 6-week periods. Each test oil served as the sole cooking oil and contributed 23% of dietary energy or two-thirds of the total daily fat intake. Dietary myristic acid (14:0) and lauric acid (12:0) from coconut oil significantly raised all the serum lipid and lipoprotein parameters measured. Subsequent one-to-one exchange of 7% energy between 16:0 (palm olein diet) and 18:1 (olive oil diet) resulted in identical serum total cholesterol (192, 193 mg/dl), low-density lipoprotein cholesterol (LDL-C) (130, 131 mg/dl), high-density lipoprotein cholesterol (HDL-C) (41, 42 mg/dl), and triglyceride (TG) (108, 106 mg/dl) concentrations. Effects attributed to gender included higher HDL in females and higher TG in males associated with the tendency for higher LDL and LDL/HDL ratios in men. However, both sexes were equally responsive to changes in dietary fat saturation. The results indicate that in healthy, normocholesterolemic humans, dietary 16:0 can be exchanged for 18:1 within the range of these fatty acids normally present in typical diets without affecting the serum lipoprotein cholesterol concentration or distribution. In addition, replacement of 12:0 + 14:0 by 16:0 + 18:1, but especially 16:0 or some component of palm olein, appeared to have a beneficial impact on an important index of thrombogenesis, i.e., the thromboxane/prostacyclin ratio in plasma.

A study on the optimal dose of aspirin therapy in Kawasaki disease--clinical evaluation and arachidonic acid metabolism.

Aspirin is the basic treatment for Kawasaki disease, however its optimal dose is controversial. We investigated the therapeutic efficacy of high-dose (100 mg/kg/day, n = 30) versus low-dose (30 mg/kg/day, n = 30) aspirin. Duration of fever, transaminase, plasma thromboxane B2 (TxB2) and 6-keto-prostaglandin F1 alpha (PGF1 alpha) levels were compared before enrollment and on days 4, 7 and 14. In the high-dose group, duration of fever was significantly shorter than that of low-dose group (3.2 +/- 1.8 versus 5.4 +/- 4.3 days, p less than 0.05), however, serum glutamic pyruvic transaminase levels were elevated (157.4 +/- 187.7 versus 48.0 +/- 58.2I.U./liter, p less than 0.005). No differences in the incidence of coronary artery lesions were observed (5 of 30 versus 7 of 30). Plasma TxB2 production was completely blocked in both groups, plasma 6-keto-PGF1 alpha levels in the high-dose group on day 14 was lower than that in the low-dose group (39 +/- 26 versus 160 +/- 207 pg/ml, p less than 0.05). This latter observation suggest that high-dose therapy may be disadvantageous as anti-thrombotic treatment, and supports the notion that low dose therapy is safe in the acute stage of Kawasaki disease.

Effects of low-dose marine oils on intimal hyperplasia in autologous vein grafts.

The effects of low-dose cod-liver oil on intimal hyperplasia of vein grafts were examined in 45 adult mongrel dogs undergoing peripheral arterial reconstruction. Fifteen animals served as the control group, 15 animals were fed a fish-oil supplement containing 240 mg of eicosapentaenoic acid daily, and a further 15 animals received 480 mg of eicosapentaenoic acid daily. Segments of undistended external jugular vein were anastomosed to bilaterally divided femoral arteries. The grafts were harvested at 6 weeks and intimal thickness was measured with a computerized interactive image analyzing system. Serum cholesterol level, prothrombin time, partial thromboplastin time, bleeding time, and platelet counts were measured before the operation and at 2, 4, and 6 weeks after the operation. Plasma levels of thromboxane B2 and prostaglandin F1 alpha and serum levels of eicosapentaenoic acid were measured before and 4 weeks after the operation. Serum cholesterol level increased similarly and significantly in all animals. Serum levels of eicosapentaenoic acid rose proportionately with the oral ingestion of fish oil but did not affect coagulation parameters. Plasma thromboxane B2 and prostaglandin F1 alpha were not significantly affected by the ingestion of marine oils. Intimal thickness was 39 +/- 5 microns in the control dogs. Ingestion of 240 mg of eicosapentaenoic acid reduced intimal thickness to 24 +/- 3 microns at 6 weeks (p less than 0.01). Increasing the dose by a factor of 2 did not decrease intimal thickness further, the intima being 23 +/- 2 microns (p less than 0.005). Our data indicate that small doses of fish oil will reduce intimal proliferation in autologous vein grafts and that marine oils may exert their beneficial effects on intimal hyperplasia by a mechanism other than their known effects on prostanoid metabolism.

Evening primose oil and fish oil are ineffective as supplementary treatment of bronchial asthma.

The effect of daily dietary supplementation with 15 to 20 mL of evening primrose seed oil or fish oil was assessed by comparison with olive oil as placebo in a cross-over study in 29 asthmatics. During 10 weeks of each regimen, the patients kept record of symptoms, peak expiratory flow rates and medication. Plasma and urine TxB2, PGE2, PGF2 alpha and 6 keto-PGF1 alpha and plasma fatty acid composition of plasma cholesterol esters were measured at the end of each treatment period. There were no differences between regimes with regard to peak flow rates, symptoms, or drug consumption. Plasma PGE2 levels increased during the fish oil treatment but there were no changes in other prostanoids in plasma or urine. The fatty acid pattern of plasma cholesterol esters showed significant differences between the supplementation periods. We conclude that moderate doses of evening primrose oil or fish oil are ineffective as a supplementary treatment of bronchial asthma.

The effect of dietary supplementation with fish oil fatty acids on surgically induced endometriosis in the rabbit.

As a means of assessing the effects of natural inhibition of cyclooxygenase enzymes on arachidonic acid metabolism in vivo, the authors supplemented the diet of 38 New Zealand white rabbits with fish oil containing eicosapentaenoic acid and docosahexaenoic acid (EPA/DHA) or olive oil (control). Endometriosis was surgically induced 10 days later using a previously described experimental technique. Peritoneal fluid PGE2 and PGF2-alpha concentrations were significantly lower in the EPA/DHA group versus controls (P less than 0.05, P = 0.05, respectively). Total endometrial implant diameter 8 weeks after induction of endometriosis was significantly smaller in the experimental group (3.1 +/- 0.2 cm) compared with the controls (4.0 +/- 0.3 cm) (P less than 0.03). The authors conclude that dietary supplementation with fish oil, containing the n-3 polyunsaturated fatty acids EPA and DHA, can decrease intraperitoneal PGE2 and PGF2-alpha production and retard endometriotic implant growth in this animal model of endometriosis.

Liposyn infusion increases plasma prostaglandin concentrations.

To determine whether Liposyn infusion results in increased plasma prostaglandin (PG) concentrations, the following study was performed in 33 adult rabbits with chronically implanted arterial and venous catheters. Plasma PG concentrations were determined by radioimmunoassay for two vasodilators, PGE2 and PGI2 (as measured by its metabolite 6-keto-PGF1 alpha), and two vasoconstrictors, thromboxane (TX) A2 and PGF2 alpha, as measured by their metabolites TXB2 and PGF2 alpha-M, respectively. A 1-hour infusion of Liposyn at 4 ml per kg resulted in statistically significant increases in arterial and venous concentrations of PGE2 and 6-keto-PGF1 alpha (p less than 0.001) and of TXB2 (p less than 0.04). There were no significant changes in PGF2 alpha-M plasma concentrations. Liposyn infusion also resulted in a small but statistically significant increase in PaO2 of 4.7 +/- 1.5 torr (p less than 0.01). It is concluded that Liposyn infusion results in statistically significant increases in plasma concentrations of PGE2, 6-keto-PGF1 alpha, and TXB2.

[Effect of trihydroxyoctadecadienoic acids on the blood content of prostaglandins E2 and F2 alpha and 5-hydroxyeicosatetraenoic acid in rats with alloxan diabetes]. / Vliianie trigidroksioktadekadienovykh kislot na soderzhanie prostaglandinov E2, F2 al'fa i 5-gidroksiéikozatetraenovoi kisloty v krovi krys pri alloksanovom diabete.

It has been demonstrated that the level of prostaglandin F2 alpha and 5-hydroxyeicosatetraenoic acid (5-HETE) in the serum of alloxan-diabetic rats is reduced by 85% and 25%, respectively, whereas that of prostaglandin E2 is increased by 34%. The administration of trihydroxyoctadecadienoic acids, that have a hypoglycemic effect, to diabetic animals brings about a rise in the level of prostaglandins F2 alpha, E2 and 5-HETE by 33%, 64% and 279%, respectively, as compared to the control.

Correction of linoleic acid deficiency in cystic fibrosis.

To identify evidence of essential fatty acid deficiency, we screened 64 patients with cystic fibrosis by analyzing total lipid extracts from plasma. Forty-three had an abnormal linoleate (18:2) level (less than 26%). Thirteen deficient patients (aged 10-24 yr) ingested for 1 yr 7% of their total calories as linoleate derived from a daily supplement of Microlipid. Five deficient patients (aged 10-37 yr) served as controls. Plasma and erythrocyte fatty acid composition were monitored by gas chromatography of total lipid extracts seven times during the twelve month period. Prostaglandins E2 and F2 alpha and their 15 keto 13, 14 dihydrometabolite, 6-keto F1 alpha, and thromboxane B2 were measured by radioimmunoassay. Sweat tests, oxygen saturation, growth indices, clinical severity scores, compliance, and possible side effects from taking Microlipid were followed. Results showed that oral supplementation with Microlipid can significantly increase plasma and erythrocytes % 18:2. One compliant patient died during the study and had normal tissue 18:2 levels. Nine of 13 patients gained more weight while taking Microlipid than in the previous year. No significant changes in sweat electrolytes, clinical scores, or oxygen saturation were found during the study year. Prostaglandin metabolites prostaglandin E2 showed an upward trend in supplemented patients, compared to controls. Prostaglandin F2 alpha remained unchanged over 1 yr but showed a trend significantly downward over the final 6 months in supplemented patients. We conclude that linoleate deficiency can be corrected with daily Microlipid supplements and that correction may alter prostaglandin metabolism.

The effect of cervical dilatation by laminaria on the plasma levels of 15-keto-13,14-dihydro-PGF2 alpha.

Laminaria tents were inserted to induce cervical dilatation prior to suction abortion in 42 primigravidae. The plasma level of 15-keto-13,14-dihydro-PGF2 alpha, the principal metabolite of prostaglandin F 2 alpha, increased during the dilatation period. The gentle dilatation by laminaria tents probably induces an endogenous synthesis of prostaglandins causing a softening of the cervix.

PIP: Laminaria tents were inserted to induce cervical dilatation prior to suction abortion in 4i primigravidae. The plasma level of 15-keto-13,14-dihydro-prostaglandin F2alpha, the principal metabolite of prostaglandin F2alpha, increased during the dilatation period. The gentle dilation by laminaria tents probably induces an endogenous synthesis of PGs causing a softening of the cervix.

Effect of zinc deficiency and food restriction on the plasma levels of prostaglandin metabolites in male rats.

The pathological signs of zinc and essential fatty acid deficiencies are highly analogous and aspirin, an inhibitor of prostaglandin synthesis, produces similar pathology when fed at toxic levels. To investigate the possibility that impaired prostaglandin biosynthesis is involved in the etiology of zinc deficiency pathology, the plasma concentrations of three prostaglandin metabolites were determined in zinc-deficient and control rats. Immature male rats were fed a purified diet low in zinc (less than 1 ppm) for 3 weeks. Ad libitum- and pair-fed controls consumed a similar diet supplemented with 100 ppm zinc. The zinc-deficient rats had low plasma zinc and their plasma concentrations of 13,14-dihydro-15-keto PGF2 alpha and 13,14-dihyro-15-keto PGE2 were significantly higher than those of the pair-fed controls. Plasma 6-keto-PGF1 alpha was not different between the zinc-deficient and pair-fed groups, but was significantly lower in deficient rats than in ad libitum-fed controls. The pair-fed controls had significantly lower plasma concentrations of all three metabolites than did the ad libitum-fed control group. The results suggest no impairment of prostaglandin production in zinc-deficient male rats.