• Archives of Pharmacal Research
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• 제28권7호
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• Toxicological Research
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• 제34권2호
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• pp.151-162
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• 2018

Anti-diabetes activity of Catharsius molossus (Ca, a type of dung beetle) glycosaminoglycan (G) was evaluated to reduce glucose, creatinine kinase, triglyceride and free fatty acid levels in db mice. Diabetic mice in six groups were administrated intraperitoneally: Db heterozygous (Normal), Db homozygous (CON), Heuchys sanguinea glycosaminoglycan (HEG, 5 mg/kg), dung beetle glycosaminoglycan (CaG, 5 mg/kg), bumblebee (Bombus ignitus) queen glycosaminoglycan (IQG, 5 mg/kg) and metformin (10 mg/kg), for 1 month. Biochemical analyses in the serum were evaluated to determine their anti-diabetic and anti-inflammatory actions in db mice after 1 month treatment with HEG, CaG or IQG treatments. Blood glucose level was decreased by treatment with CaG. CaG produced significant anti-diabetic actions by inhiting creatinine kinase and alkaline phosphatase levels. As diabetic parameters, serum glucose level, total cholesterol and triglyceride were significantly decreased in CaG5-treated group compared to the controls. Dung beetle glycosaminoglycan, compared to the control, could be a potential therapeutic agent with anti-diabetic activity in diabetic mice. CaG5-treated group, compared to the control, showed the up-regulation of 48 genes including mitochondrial yen coded tRNA lysine (mt-TK), cytochrome P450, family 8/2, subfamily b, polypeptide 1 (Cyp8b1), and down-regulation of 79 genes including S100 calcium binding protein A9 (S100a9) and immunoglobulin kappa chain complex (Igk), and 3-hydroxy-3-methylglutaryl-CoenzymeAsynthase1 (Hmgcs1). Moreover, mitochondrial thymidine kinase (mt-TK), was up-regulated, and calgranulin A (S100a9) were down-regulated by CaG5 treatment, indicating a potential therapeutic use for anti-diabetic agent.

• Natural Product Sciences
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• 제8권2호
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• pp.66-70
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• 2002

The in vitro anticoagulant and fibrinolytic activities of crude extracts from insects were evaluated in order to find effective therapeutic drugs for the treatment of myocardial and cerebral thrombosis. We prepared three types of extracts (water, methanol and ethylacetate) from 28 insects for use as raw materials for the activity assays. The fibrinolytic activity was tested using the fibrin plate method and the activated partial thromboplastin time and thrombin time were measured for blood clotting activity. With regards to the fibrinolytic system, water extracts of six kinds of insects displayed a remarkable level of activity with a plasmin-like action. The water extracts of [Catharsius molossus, Eupolyphaga sinensis, Huechys sanguinea, Mantidis $o\ddot{o}theca$, Mimela splendens, and Polistes mandarinus (Vespae Nidus)] exhibited the activity. On the other hand, the methanol extracts did not display any fibrinolytic activity. In terms of the coagulation system, an aqueous extract of silkworm Tongchunghacho (Paecilomyces japonica), Oxya japonica japonica and Buthus martensi (Scorpion) increased the clotting time significantly longer (181 times) than the control. These results suggest that crude drugs from insects are useful sources for the development of new drugs for use in treatments involving blood coagulation and fibrinolysis.

• 생명과학회지
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• 제26권7호
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• pp.772-781
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• 2016

쇠똥구리는 동물의 분을 이용하여 토질을 개선시켜 지구 생태계에 매우 중요한 역할을 담당하는 곤충이다. 이 연구에서 수컷 SD rat는 5군으로 분리되어 PBS, 쇠똥구리 에탄올 추출물, 쇠똥구리 아세톤 추출물, 귀뚜라미 아세톤 추출물, 양성대조군으로 새우기름 그룹으로 나누었다. 13주령 쥐에 고지방식이로 비만을 7주간 유도하고, 20주령부터 시험물질을 고지방 식이를 하면서 한 달간 투여하였다. 쇠똥구리 추출물의 투여는 체중과 복부지방과 부고환지방 중량의 감소로 이어졌다. 지질의 산화적 스트레스는 간에서 MDA를 측정하여 평가하였으나, 군간에 유 의성이 없었다. 단백질의 산화적 스트레스는 혈액에서 단백질 카르보닐 양으로 측정되었는데 쇠똥구리 에탄올 추출물과 쇠똥구리 아세톤 추출물에서 유의성있게 산화적 스트레스가 감소되었다. 반면에 간에서의 단백질 카르보닐 양은 군간에 유의성이 없었다. 당뇨 혈관내피세포를 효소면역분석법을 사용하여 세포부착단백질인 laminin 과 fibronectin 수준을 측정하였더니 쇠똥구리 추출물에서 유의성이 있었다. 사이토카인 IL-10, IL-1β, VEGF, eNOS를 측정하였는데 IL-10에서 쇠똥구리 에탄올 추출물과 쇠똥구리 아세톤 추출물에서 유의성이 있었다. 항산 화효소인 SOD, GSH-Px 활성은 군간에 유의성은 없었지만, 증가하는 경향이 있었고, CAT활성은 쇠똥구리 추출물에서 유의성있게 증가되었다. 지방조직 내의 포화지방산 비율이 감소하는 경향이 나타났으며, 불포화지방산과 다 가불포화지방산은 증가하였다. 쇠똥구리 추출물의 지방조직 감소 및 여러 지질 수치의 향상효과는 쇠똥구리 추출물을 기능성 곤충 소재로 이용할 수 있는 가능성을 제시한다.