• Biomedical Science Letters
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• v.29 no.1
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• pp.41-47
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• 2023

Discovery of new substance that can regulate platelet aggregation or suppress aggregation will aid in the prevention and treatment of cardiovascular diseases. Artesunate is a compound from plant roots of Artemisia or Scopolia, and its effects have shown to be promising in areas of anticancer and Alzheimer's disease. However, the role and mechanisms by which artesunate affects the aggregation of platelets, and the formation of a thrombus are currently not understood. This study examined the ways artesunate affects platelets activation and thrombus formation induced by collagen. As a result, cAMP and cGMP production were increased significantly by artesunate relative to the doses, as well as phosphorylated VASP and IP₃R, substrates to cAMP-dependent kinase and cGMP-dependent kinase, in a significant manner. The Ca²⁺ normally mobilized from the dense tubular system was inhibited due to IP₃R, phosphorylation from artesunate, and phosphorylated VASP aided in inhibiting platelet activity via αIIb/β₃ platelet membrane inactivation and inhibiting fibrinogen binding. Finally, artesunate inhibited thrombin-induced thrombus formation. Therefore, we suggest that artesunate has importance with cardiovascular diseases stemming from the abnormal platelets activation and thrombus formation by acting as an effective prophylactic and therapeutic agent.

• Microbiology and Biotechnology Letters
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• v.51 no.2
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.

• Korean Journal of Veterinary Research
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• v.37 no.2
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• pp.451-456
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• 1997

120 heads of korean native calves were examined of serum protein by using cellulose acetate electrophoresis. During 20 days since the calves were purchased, serum protein, fibrinogen values and plasma : fibrinogen ratio were examined in the calves with respiratory and diarrheal disease. The result obtained in this investigation were as follows : 1. Among the 120 heads of calves that were purchased from market, 14 heads(22%) of calves were occurred respiratory disease, and 12 heads(20%) of calves are occurred diarrhea. Occurrence of respiratory disease was 14.5(4~20) days in average and diarrhea was 9.6(2-15) days after they had been purchased. 2. Serum protein fraction were analyzed by cellulose acetate electrophoresis. ${\beta}-globulin$, A/G ratio and ${\beta}_2-globulin$ values were decreased in the calves with respiratory disease. Especially, ${\beta}_2-globulin$ were significantly decreased. In calves with diarrhea, there was no change in ${\beta}-globulin$ values. ${\beta}_2-globulin$ values were higher than that of the normal and respiratory diseased calves. 3. ${\alpha}-globulin$ values were increased in both of calves with diarrhea and respiratory disease. This tendency was due to increase ${\alpha}_2-globulin$ values. 4. The $\gamma$-globulin value of calves with diarrhea was the lowest among the 3 groups. 5. The total protein values of normal calves were $7.0{\pm}1.1g/dl$ and that of respiratory and diarrheal diseased calves were $6.9{\pm}0.9g/dl$ and $6.6{\pm}0.8g/dl$, respectively. Total protein value of calves with diarrhea was lower than that of normal and respiratory diseased calves. Globulin value of calves with diarrhea was the lowest among them. The low value of total protein in diarrheal diseased calves was due to decrease globulin values. 6. The fibrinogen values of calves with respiratory disease ($643{\pm}189mg/dl$) were significantly higher than that of normal calves($533{\pm}135mg/dl$) and calves with diarrhea($572{\pm}188mg/dl$). The plasma : fib. ratio of respiratory diseased calves was $12.0{\pm}4.9$, normal calves was $13.8{\pm}3.5$ and diarrheal diseased calves was $12.8{\pm}4.6$. The ratio of the calves with respiratory disease was significantly decreased.

• Journal of Embryo Transfer
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• v.25 no.1
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• pp.29-34
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• 2010

Follicular cystic ovary (FCO) is one of the major causes of reproductive failure in cattle. Genetic alterations affect the function of diverse cells and/or tissues, which could be present in cystic ovaries. A microarray analysis was performed to screen differential gene expressions in follicular cystic follicles of cattle. In this study, we hypothesized that follicular cysts may be induced by changes in ion- and transporter-related gene expression. Microarray data showed that fibrinogen-gamma (FGG) and low density lipoprotein receptor-related protein 8 (LRP8) were up-regulated, while choline transporter-like protein 4 (SLC44A4), very long-chain acyl-CoA synthetase homolog 2 (SLC27A5), annexin 8 (ANXA8), and aquaporin 4 were down-regulated in follicular cystic follicles. A semi-quantitative RT-PCR was carried out to validate DEGs altered in follicular cystic follicles. Of six DEGs, three DEGs (FGG, SLC44A4, and aquaporin 4) showed a positive correlation between microarray and semi-quantitative PCR data. We focused on FGG, among three DEGs, which was highly up-regulated in follicular cystic follicles. The FGG mRNA was upregulated by 8.4-fold and by 1.7-fold in the bovine follicular cystic follicles as judged by microarray and RT-PCR analysis, respectively. However, there was no significant changes in the expression level of FGG protein in both follicular cystic follicles and granulosa cells isolated from follicular cystic follicles by Western blot analysis. Although this study does not reveal a positive correlation between the mRNA and protein level, FGG appears to be an important biomarker in the discrimination of follicular cyst from normal ovary.

• The Journal of Internal Korean Medicine
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• v.19 no.1
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• pp.114-133
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• 1998

In order to investigate the effects of Alcohol-Steamed Rhei Rhizoma and Row Rhei Rhizoma on varied extract time in both endotoxin-induced blood stasis model(hereafter Endotoxin Model) and hydrocortisone acetate-induced blood stasis model (hereafter HA Model), Half of rats were treated with endotoxin(0.4mg/kg, single Ⅳ, into caudal vein) for Endotoxin Model. Thereafter, they were orally administrated water extract of Alcohol-Steamed Rhei Rhizoma or Row Rhei Rhizoma, which were boiled during 30, 60, 120 minute, respectively. Finally, the number of platelet, fibrinogen, prothrombin time, hematocrit, the number of RBC and WBC were measured after sacrifice. The remainder rats were treated with hydrocortisone acetate(10mg/kg, daily IM for 7 days into the muscular rump) for HA significantly decreased. Together, they were orally administrated for 7 days water extract of Alcohol-Steamed Rhei Rhizoma or Row Rhei Rhizoma that were boiled above methods Finally, the number of platelet, fibrinogen, prothrombin time, hematocrit, the number of RBC and WBC were measured after sacrifice. The results were summarized as follows : 1. The number of platelet was significantly increased in boiled water extract for 30 min of Row Rhei Rhizoma group as compared with that of control group in Endotoxin Model. 2. Fibrinogen level was significantly increased in all administration groups as compared with that of control group in Endotoxin Model. It was significantly increased in all administration groups except boiled water extract for 30 min of Alcohol-Steamed Rhei Rhizoma group as compared with that of control group in HA Model. 3. Prothrombin time was significantly shortened in boiled water extract for 60 min of Alcohol-Steamed Rhei Rhizoma group and boiled water extract for 120 min of Alcohol-Steamed Rhei Rhizoma group as compared with that of control group in Endotoxin Model It was significantly shortened all administration groups as compared with that of control group in HA Model. 4 Hematocrit was significantly increased in all administered groups except boiled water extract for 60 min of Alcohol-Steamed Rhei Rhizoma group and boiled water extract for 30 min of Row Rhei Rhizoma group as compared with that of control group in Endotoxin Model. It was significantly increased in all administration groups as compared with that of control group in HA Model. 5. The number of RBC was significantly decreased in boiled water extract for 60 min of Alcohol-Steamed Rhei Rhizoma group, boiled water extract for 120 min of Alcohol -Steamed Rhei Rhizoma group and boiled water extract for 30 min of Row Rhei Rhizoma administered group in Endotoxin Model. It was significantly increased boiled water extract for 30 min of Row Rhei Rhizoma group and boiled water extract for 60 min of Row Rhei Rhizoma group in HA Model as compared with data of control group. 6 The number of WBC was significantly decreased in all administered groups except boiled water extract for 30 min of Alcohol-Steamed Rhei Rhizoma group and boiled water extract for 60 min of Alcohol-Steamed Rhei Rhizoma group as compared with that of control group in HA Model.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.6
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• pp.788-794
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• 2013

Mastitis set apart as clinical and sub clinical is a disease complex of dairy cattle, with sub clinical being the most important economically. Of late, laboratories showed interest in developing biochemical markers to diagnose sub clinical mastitis (SCM) in herds. Many workers reported noteworthy alternation of acute phase proteins (APPs) and nitric oxide, (measured as nitrate+nitrite = NOx) in milk due to intra-mammary inflammation. But, the literature on validation of these parameters as indicators of SCM, particularly in riverine milch buffalo (Bubalus bubalis) milk is inadequate. Hence, the present study focused on comparing several APPs viz. ${\alpha}_1$-anti trypsin, ${\alpha}_1$-acid glycoprotein, fibrinogen and NOx as indicators of SCM in buffalo milk. These components in milk were estimated using standardized analytical protocols. Somatic cell count (SCC) was done microscopically. Microbial culture was done on 5% ovine blood agar. Of the 776 buffaloes (3,096 quarters) sampled, only 347 buffaloes comprising 496 quarters were found positive for SCM i.e. milk culture showed growth in blood agar with $SCC{\geq}2{\times}10^5$ cells/ml of milk. The cultural examination revealed Gram positive bacteria as the most prevalent etiological agent. It was observed that ${\alpha}_1$-anti trypsin and NOx had a highly significant (p<0.01) increase in SCM milk, whereas, the increase of ${\alpha}_1$-acid glycoprotein in infected milk was significant (p<0.05). Fibrinogen was below detection level in both healthy and SCM milk. The percent sensitivity, specificity and accuracy, predictive values and likelihood ratios were calculated taking bacterial culture examination and $SCC{\geq}2{\times}10^5$ cells/ml of milk as the benchmark. Udder profile correlation coefficient was also used. Allowing for statistical and epidemiological analysis, it was concluded that ${\alpha}_1$-anti trypsin indicates SCM irrespective of etiology, whereas ${\alpha}_1$-acid glycoprotein better diagnosed SCM caused by gram positive bacteria. NOx did not prove to be a good indicator of SCM. It is recommended measuring both ${\alpha}_1$-anti trypsin and ${\alpha}_1$-acid glycoprotein in milk to diagnose SCM in buffalo irrespective of etiology.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.1
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• pp.103-108
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• 1997

It has been known that garlic, one of the essential ingredients of spices in Korean food, has a hypotensive effect. The following experiments were done to compare the effect of heat treatment of garlic on change in blood pressure. We selected SHR(Spontaneously Hypertension Rat) for experimental animals since, in the case of human beings, 85~90% of high blood pressure is in hereditary origin. Animals were divided into 3 groups, control group(no garlic), 3% raw garlic group and 3% heated garlic group. Serum was analyzed for lipid concentration, and plasma for prothrombin time and fi-brinogen concentration. The effects of heat treatment of garlic were as follow. There was no significant differences in body weight gain and feed efficiency ratio except that feed intake of 3% heated garlic-fed group was significantly lower than that of control group and 3% raw garlic-fed group. Garlic-fed groups, in contrast to the control group, showed significant difference in cholesterol content in pro-thrombin time and in fibrinogen concentration. Taken together, hypotensive effects of garlic on high blood pressure were significant. Regardless of heat treatment both heated garlic and raw garlic showed hypotensive effects.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.434-438
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• 2005

In the previous study, we isolated a myulchikinase (MK), which has fibrinolytic activity and cytotoxicity to the tumor cell line, from myl- chi-jeot-gal. In this study, the effect of NaCl concentration, metallic ions, pH, temperature, and plasminogen on the activity of MK was analysed. The MK activity was maintained at least $80\%$ activity up to $30\%$ NaCl, which indicates that the enzyme may be halotolerant. The optimal pH and temperature were 8 and $40^{\circ}C$, respectively. The fibrinolytic activity of MK was completely inhibited with 0.5 mM $Hg^{2+}$ and inhibited to $50^{\circ}C$ with 1 mM $Cu^{2+}\;and\;Zn^{2+}$. The MK showed strong activity in plasminogen- rich fibrin plate but not in plasminogen-free fibrin plate. The result indicates that the MK may be a plasminogen activator type fibrinolytic enzyme.

• Journal of Life Science
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• v.24 no.8
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• pp.873-879
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• 2014

The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.

• The Journal of Korean Medicine
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• v.19 no.1
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• pp.179-204
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• 1998

The purpose of this research was to investigate the efficacy of Goiwhasan extract powder on the gastric injuries, antiulcer, gastrointestinal tract and blood hemostasis. Animals were used through this studies mice and rats. All animals were divided into 3 groups, contol group(no treatment), sample Ⅰ group(375mg/kg administration), sample Ⅱ group(750mg/kg administration). The gastric injuries and ulcer have been made by using pyloric ligation, indomethacin, HCI-ethanol, acetic acid and then The histological observation was followed. In the gastrointestinal tract, gastric juice secretion, gastric acidity, pepsin output, blood gastrin and secretin level, transport potentials in the small and large intestine were checked. And studies on blood hemostasis were performed on normal hemostatic activities and plasma prothrombin time, plasma recalcification time, plasma fibrinogen levels in the hypoprothrombinemic mice induced by warfarin. The results were as follows: 1. The antigastric ulcer effects on the pyloric ligation, indomethacin, HCl-ethanol, acetic acid induced gastric injuries were shown in Sample Ⅱ group(p<0.05). 2. Through the morphologic examination on the acetic acid induced ulcer, Sample Ⅰ group showed mild regeneration of epithelium and slight decrease of periulcer edema then that of Control group, while Sample Ⅱ group showed more retraction of round ulcer site, remarkable loss of swelling and edema then that of Control group, and revealing the regenerated epithelium in the surrounding ulcer site. Thus it was noted that both Sample groups have antigastriculcer effects on the experimentally induced gastric ulcer. 3. The inhibitory effects on gastric juice were noted in both Sample Ⅰ group(p<0.05) and Sample Ⅱ group(p<0.01). However, only Sample Ⅱ group showed the inhibitory effects on total acidity and pepsin output(p<0.05). 4. The significant inhibition of blood gastrin level showed at 30 min.(P<0.05) and 90 min.(P<0.05) after starting medication in only Sample Ⅱ group, but significance of blood secretin level in both groups was not recognized. 5. Any significant changes in barium sulfate transport in the small intestine of mice was not recognized in both groups, but the significantly inhibitory effect in large intestine was recognized in both Sample Ⅰ group(p<0.05) and Sample Ⅱ group(p<0.001). 6. In hemostatic effect on both normal mice and hypoprothrombinemic mice induced by warfarin, the significantly shortening effect on coagulation time was seen in only Sample Ⅱ group(p<0.01). 7. On plasma prothrombin time in hypoprothrombinemic rat induced by warfarin, Sample Ⅱ group have shortened the prothrombin time significantly(p<0.001). 8. On plasma reclcification time in hypoprothrombinemic rat induced by warfarin, the recalcification time have been shortened significantly in both Sample Ⅰ group(p<0.05) and Sample Ⅱ group(p<0.01). 9. On plasma fibrinogen levels in hypoprothrombinemic rat induced by warfarin, the fibrinogen contents in Sample Ⅱ have been decreased significantly(p<0.01). Overall the above results suggest that Goiwhasan has an therapeutic efficacy on antigastric ulcer and blood hemostasis. Further studies would be needed on the interaction of its herbal medicine and its mechanism in the future.