[The role of angelica injection on P-selectin and anti-cardiolipin antibodies in acute pulmonary embolism rats].

OBJECTIVE: To study the effect of Chinese medicine, Angelica, injection on the expression of P-, E-selectin and anti-cardiolipin antibody in acute pulmonary embolism rats. METHODS: SD rats were randomly divided into 3 groups: normal control group(Group N), thromboembolism group (Group T), and treatment group of thromboembolism with angelica injection (Group TA). There were three time points in every group: 1 h, 4 h and 8 h. Plasma was detected by P-, with 4% paraformaldehyde, and paraffin embedded sections were detected by immunohistochemistry for P-, E-selectin and anti-cardiolipin antibodies. RESULTS: With HE stain, the inflammatory cells in the lung of rats were relatively rare in every time point in normal control group. In group T and group TA, the inflammatory cells were increasing in every time point in comparison to group N (P < 0.05) and the inflammatory cells were increasing with time in group T. The data revealed that the plasmic level of P-, E-selectin was significantly higher than that in group T1, group T4, group T8 in comparison to the corresponding sub groups of group N (P < 0.05), while it was significantly lower than that in group TA1, group TA4, group TA8 in comparison to the corresponding sub groups of group T (P < 0.05); For the OD value of plasmic anti-cardiolipin antibodies (ACA), no significant difference was observed during was lower expressed by immunohistochemistry. CONCLUSION: Acute pulmonary embolism can lead to infiltration of inflammatory cell in rat lungs. The lung inflammation of acute pulmonary embolism rats can be enhanced probably by the increased release of P-, E-selectin and anti-cardiolipin antibodies, and the enhanced inflammation promotes the release of a series of inflammatory mediators, which exacerbate the injury of lung. Angelica injection relieves the lung inflammation of acute pulmonary embolism rats possibly by inhibiting the expression of P-, E-selectin and anti-cardiolipin antibody, thus playing a role in reducing thrombogenesis.

Blockade of glycoprotein IIb/IIIa mediates the antithrombotic activity of butanol fraction of Actinostemma lobatum Maxim.

AIM OF THE STUDY: Actinostemma lobatum Maxim, a wildlife plant of Cucurbitaceae family, has been utilized for the prevention or treatment of cardiovascular diseases as a folk remedy in Korea. However, its scientific evidence remains unclear. Thus, in the present study, we examined the effects of butanol fraction of Actinostemma lobatum Maxim (BFALM) on the in vitro and in vivo antithrombotic activity and possible mechanisms were elucidated for the first time. MATERIAL AND METHODS: To elucidate the antithrombotic mechanism of BFALM, platelet aggregation assay, coagulation assay, glycoprotein IIb/IIIa assay, thromboxane A(2) assay and in vivo pulmonary thromboembolism experiment were performed. RESULTS: BFALM significantly inhibited collagen, adenosine diphosphate (ADP) and thrombin-induced platelet aggregation in a concentration dependent manner. Consistently, oral administration of BFALM resulted in a dose-dependent increase of survival rates of mice with pulmonary thromboembolism induced by intravenous injection of collagen and epinephrine. In mechanism assays for the antithrombotic activity of BFALM, BFALM significantly inhibited the fibrinogen binding to the platelet surface Glycoprotein IIb/IIIa (GP IIb/IIIa) receptor in a concentration dependent fashion, as well as reduced the level of thromboxane A(2) at 400microg/ml. Furthermore, BFALM significantly prolonged the prothrombin time (PT) and activated partial thromboplastin time (APTT) compared with untreated control. CONCLUSIONS: These results suggest that BFALM may exert antithrombotic activity through inhibition of platelet aggregation via GP IIb/IIIa and thromboxane A(2) pathways, along with anticoagulatory activity through intrinsic and extrinsic pathways.

GPVI-deficient mice lack collagen responses and are protected against experimentally induced pulmonary thromboembolism.

Platelet glycoprotein VI (GPVI) is now considered to be a major player in platelet-collagen adhesive interactions leading to thrombus formation. GPVI blockade, or its depletion, has been shown in mice to result in complete protection against arterial thrombosis, without significant prolongation of bleeding time. GPVI may therefore represent a useful antithrombotic target. In order to reaffirm the role of GPVI in platelet-collagen interactions, we developed GPVI(null) mice by targeted disruption methodology. GPVI(null) mice platelets failed to respond to a high dose of fibrillar collagen, or convulxin, a GPVI agonist, but showed a normal response to other agonists such as ADP, PMA and arachidonic acid. We report, for the first time, that a proportion of GPVI(null) mice is protected against lethal thromboembolism, induced by the infusion of a mixture of collagen and epinephrine. Greater than 55% of GPVI(null) mice survived the challenge, whereas the maximal survival from the other genotypes was 17% (n=18 per genotype). Washed platelets obtained from GPVI(null) mice showed >90% reduction in adhesion to fibrillar collagen under static conditions. Platelet adhesion to collagen under dynamic conditions using a high shear rate (2600 s(-1)) was dramatically reduced using blood from GPVI(null) mice, while platelets from wild-type and heterozygous animals showed a similar amount of adhesion. Animals from each genotype had essentially similar tail bleeding time, suggesting that a complete deficiency of GPVI, at least in mice, does not result in an enhanced bleeding tendency. These observations clearly establish that blockade of GPVI may attenuate platelet-collagen interactions without adversely affecting the bleeding time.

Demonstration of enhanced endogenous fibrinolysis in thrombin activatable fibrinolysis inhibitor-deficient mice.

To investigate the importance of thrombin activatable fibrinolysis inhibitor (TAFI) in the stabilization of plasma clots, we have compared fibrinolysis in TAFI-deficient (KO) and wild-type (WT) littermate mice. TAFI-deficient mice were previously generated by targeted gene disruption. The level of TAFI activity generated in plasma from WT mice in the presence of added thrombin and thrombomodulin (activatable TAFI) is twice that of plasma from TAFI heterozygous mice (HET); no activatable TAFI is detected in TAFI KO plasma. In vitro, TAFI KO plasma clots lysed faster than WT plasma clots, and HET plasma clots lysed at an intermediate rate. The rate of clot lysis for KO mice is not changed in the presence of potato carboxypeptidase inhibitor, a specific inhibitor of TAFIa, whereas the WT and HET clot lysis rates are increased in the presence of potato carboxypeptidase inhibitor. C-terminal lysine residues are preserved on partially degraded clots from KO mice, but are absent from partially degraded WT clots. In vivo, in a batroxobin-induced pulmonary embolism model, KO mice displayed a lower retention of fibrin in the lungs than did WT mice. These results are the first demonstration of enhanced endogenous fibrinolysis in an in vivo model without the addition of exogenous thrombolytic.

Effects of short-term, high dose, heparin therapy on biochemical markers of bone metabolism.

To evaluate the effects of the short-term, high-dose sodium heparin therapy on biochemical markers of bone metabolism, we studied 20 patients (11 males and 9 females) with pulmonary embolism, treated with sodium heparin (daily dose range: 40,000-45,000 I.U. by continuous i.v. infusion). Heparin therapy lasted 5-7 days, after which patients received warfarin over 12 months. Eleven patients (6 males and 5 females) with ischaemic stroke, treated with i.v. glycerol and pentoxifilline, were used as controls. Before and after therapy serum and urinary markers of bone metabolism were evaluated; in 12 heparin-treated pts., the parameters were also evaluated 4 months after discontinuation of warfarin therapy. After heparin therapy a significant reduction vs. basal value was observed in levels of serum osteocalcin (ng/ml;mean + SEM): 3.32 & 0.19 vs. 2.05 + 0.21; p < 0.001. In the 12 patients evaluated 4 months after discontinuation of warfarin therapy, serum osteocalcin levels returned to basal value: 3.41 + 0.12 ng/ml (p:n.s.). No significant changes of the examined parameters were observed in controls. In conclusion, our data seem to indicate an effect of i.v. short-term heparin therapy on bone metabolism. This effect seems to be characterized by an inhibition of osteoblast function as suggested by the reduction of serum osteocalcin levels.

Pulmonary transvascular fluid and protein exchange after thrombin-induced microembolism. Differential effects of cyclooxygenase inhibitors.

We studied the effects of cyclooxygenase inhibitors (indomethacin, meclofenamate, and ibuprofen) on the increases in pulmonary transvascular fluid and protein fluxes that occur after thrombin-induced intravascular coagulation. Each drug-pretreated group was compared with a control group receiving the same amount of alpha-thrombin (alpha-T), the native enzyme used to induce intravascular coagulation. Studies were made in the following groups of anesthetized sheep prepared with lung lymph fistulas: low dose-indomethacin (bolus injection of 5 mg/kg and infusion of 3 mg/kg/h, high-dose indomethacin (bolus injection of 20 mg/kg and infusion of 12 mg/kg/h), meclofenamate (bolus injection of 5 mg/kg and infusion of 3 mg/kg/h), low-dose ibuprofen (bolus of 6.2 mg/kg and infusion of 3.1 mg/kg/h), and high-dose ibuprofen (bolus of 25 mg/kg and infusion of 12.5 mg/kg/h). The concentrations of the arachidonic acid metabolites, thromboxane B2 and 6-keto-prostaglandin F1 alpha, did not increase in the treated groups. Infusion of alpha-T increased the mean pulmonary arterial pressure and pulmonary vascular resistance in the control and treated groups, but to higher levels in the indomethacin and meclofenamate groups. In the control group, alpha-T increased pulmonary lymph flow (Qlym) and lymph protein clearance (Qlym X lymph/plasma protein concentration ratio), a measure of lung vascular permeability. In the indomethacin, meclofenamate, and low-dose ibuprofen groups, alpha-T resulted in delayed increases in Qlym and lymph protein clearance, but these lymph parameters attained the same levels as the control groups. However, the high-dose of ibuprofen significantly attenuated the increases in Qlym and lymph protein clearance after alpha-T.(ABSTRACT TRUNCATED AT 250 WORDS)

[Cytophotometric study of the DNA concentration in the supraoptic neurosecretory nuclei of the hypothalamus]. / Tsitofotometricheskoe issledovanie soderzhaniia DNK v supraopticheskikh neirosekretornykh iadrakh gipotalamusa.

DNA content and distribution in supraoptic neuronal nuclei and in their satellites was studied in human subjects died under different conditions of hypothalamo-hypo-physeal neurosecretory activity: moderate (control) and high. In control observations, prevalence of diploid and paradiploid nuclei both in the secretory neurons and in the nuclei of glial satellites was noted. High neurosecretory activity was connected with a tendency towards increased DNA content in the neuronal nuclei, up to the appearance of some tetraploid elements. In the glial nuclei of the satellites, events of poliploidization were observed, that is, evidently, of adaptive character to maintain active functioning of the neurons under certain intensified conditions.