DLBS1033, a protein extract from Lumbricus rubellus, possesses antithrombotic and thrombolytic activities

Jessica Trisina, Febrina Sunardi, Maggy T Suhartono, Raymond R Tjandrawinata, Jessica Trisina, Febrina Sunardi, Maggy T Suhartono, Raymond R Tjandrawinata

Abstract

The medicinal value of earthworm has been widely known since the history of Asian ancient medicine. This present study aims to determine the mechanism of action and effect of a standardized extract of Lumbricus rubellus named as DLBS1033. The fibrinogen degradation, antiplatelet aggregation, and ex vivo antithrombotic assay using human blood were performed to study antithrombotic activity. Fibrin plate and clot lysis assay were also done to examine thrombolytic properties. DLBS1033 was found to possess fibrinogenolytic activity on α-, β-, and γ-chain of fibrinogen. It also induced antiplatelet aggregation and prolonged blood clotting time, which further confirmed its antithrombotic properties. In addition, thrombolytic properties of DLBS1033 were shown with its fast and long-acting fibrinolytic activity, as well as its effective blood clot lysis activities. In conclusion, DLBS1033 conferred antithrombotic and thrombolytic action which could be used as a safe and promising oral thrombolytic drug.

Figures

Figure 1
Figure 1
Stability of DLBS1033 in the range of pH. DLBS1033 was examined in pH range of 2–14.
Figure 2
Figure 2
Stability of DLBS1033 in the range of temperature. DLBS1033 was examined in temperature range of 25°C–80°C for 5 hours.
Figure 3
Figure 3
Lumbricus low-molecular-weight protein profile of DLBS1033. The eathworm extract has 8 major proteins with molecular weight below 100 kDa.
Figure 4
Figure 4
Fibrinogen zymogram of DLBS1033. LLP fractions which contribute to fibrinogenolytic activity were identified as clear bands.
Figure 5
Figure 5
Activity of DLBS1033 on chromozym substrate. (a) Change in the absorbance at 405 nm during assay of DLBS1033 with chromozyme. (b) Linear section of changes in absorbance per one minute.
Figure 6
Figure 6
Fibrinogen degradation by DLBS1033. Lane 1: fibrinogen. Lanes 2–8 representing mixtures aliquot were taken at 0, 5, 15, 30, 45, 60, and 90 min, respectively.
Figure 7
Figure 7
Inhibition of platelet aggregation by DLBS1033. Platelet aggregation induced by thrombin (a) and ADP (b) was downregulated by the administration of DLBS1033. EDTA and cilostazol were used as the positive control.
Figure 8
Figure 8
Ex vivo antithrombotic effect of DLBS1033. The clot formation was inhibited after the administration of DLBS1033. +++: fully clotted; high clotting consistency; ++: partially clotted, medium clotting consistency; +: slightly clotted, low clotting consistency; −: no clot occurred.
Figure 9
Figure 9
Volume of fibrin lysed by DLBS1033 was calculated at incubation time intervals.
Figure 10
Figure 10
Lysis of blood clot by DLBS1033. Clot lysis was measured and compared between control and DLBS1033.

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