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Nattokinase Decreases Plasma Levels of Fibrinogen, Factor VII, and Factor

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Nattokinase Decreases Plasma Levels of Fibrinogen, Factor VII, and Factor Available online at www.sciencedirect.com Nutrition Research 29 (2009) 190–196 www.nrjournal.com Nattokinase decreases plasma levels of fibrinogen, factor VII, and factor VIII in human subjects☆ Chien-Hsun Hsiaa, Ming-Ching Shenb, Jen-Shiou Linc, Yao-Ke Wend, Kai-Lin Hwange, ⁎ Thau-Ming Chamf, Nae-Cherng Yangg, aDivision of Cardiovascular Medicine, Changhua Christian Hospital, Changhua 500, Taiwan bDivision of Hematology-Oncology, Changhua Christian Hospital, Changhua 500, Taiwan cDepartment of Laboratory Medicine, Changhua Christian Hospital, Changhua 500, Taiwan dDivision of Nephrology, Changhua Christian Hospital, Changhua 500, Taiwan eDepartment of Public Health, Chung Shan Medical University, Taichung 402, Taiwan fGraduate Institute of Pharmaceutical Sciences, Kaohsiung Medical University, Kaohsiung 807, Taiwan gDepartment of Nutrition and Health Science, Chungchou Institute of Technology, Changhua 510, Taiwan Received 17 December 2008; revised 21 January 2009; accepted 28 January 2009 Abstract Nattokinase, a serine proteinase from Bacillus subtilis, is considered to be one of the most active functional ingredients found in natto. In this study, we hypothesized that nattokinase could reduce certain factors of blood clotting and lipids that are associated with an increase risk for cardiovascular disease (CVD). Thus, an open-label, self-controlled clinical trial was conducted on subjects of the following groups: healthy volunteers (Healthy Group), patients with cardiovascular risk factors (Cardiovascular Group), and patients undergoing dialysis (Dialysis Group). All subjects ingested 2 capsules of nattokinase (2000 fibrinolysis units per capsule) daily orally for 2 months. The laboratory measurements were performed on the screening visit and, subsequently, regularly after the initiation of the study. The intent-to-treat analysis was performed on all 45 enrolled subjects. By use of mixed model analysis, a significant time effect, but not group effect, was observed in the change from baseline of fibrinogen (P = .003), factor VII (P b .001), and factor VIII (P b .001), suggesting that the plasma levels of the 3 coagulation factors continuously declined during intake; also, the extents of decrease were similar between groups. After 2 months of administration, fibrinogen, factor VII, and factor VIII decreased 9%, 14%, and 17%, respectively, for the Healthy Group; 7%, 13%, and 19%, respectively, for the Cardiovascular Group; and 10%, 7%, and 19%, respectively, for the Dialysis Group, whereas blood lipids were unaffected by nattokinase. No significant changes of uric acid or notable adverse events were observed in any of the subjects. In summary, this study showed that oral administration of nattokinase could be considered as a CVD nutraceutical by decreasing plasma levels of fibrinogen, factor VII, and factor VIII. © 2009 Elsevier Inc. All rights reserved. Keywords: Nattokinase; Cardiovascular disease; Human; Fibrinogen; Factor VII; Factor VIII; Cholesterol Abbreviations: CVD, cardiovascular disease; FU, fibrinolysis unit; HDL-C, high-density lipoprotein cholesterol; LDL-C, low- density lipoprotein cholesterol; LSD, least significant difference; TC, total cholesterol; TG, triacylglycerol. ☆ The Research Fund of Changhua Christian Hospital supported this study (Project No I050308). ⁎ Corresponding author. Tel.: +886 4 8395316; fax: +886 4 8395316. E-mail address: [email protected] (N.-C. Yang). 0271-5317/$ – see front matter © 2009 Elsevier Inc. All rights reserved. doi:10.1016/j.nutres.2009.01.009 C.-H. Hsia et al. / Nutrition Research 29 (2009) 190–196 191 1. Introduction [21]. Hence, data on uric acid concentration and adverse events collected via a self-administered questionnaire were Nattokinase, a serine proteinase from Bacillus subtilis, used to monitor the safety of nattokinase administration. has been reported to have potent fibrinolytic activity [1]. The enzyme is composed of 275 amino acids with a molecular 2. Methods and materials weight of 27.7 KDa in its mature form [2] and is considered to be one of the most active functional ingredients found in 2.1. Subjects natto, a traditional Japanese food prepared from fermented soybeans. Several studies in vitro have shown that This study used an open-label, self-controlled design. nattokinase has potent fibrinolytic activity [2-4]. In rat Study protocols and materials were reviewed and approved studies, nattokinase was found to have an effect approxi- by the institutional review board of Changhua Christian mately 4 times stronger than plasmin in dissolving a Hospital, Taiwan, before initiating the study. Forty-five thrombus [1,5]. A small trial on 12 adult Japanese subjects adult men and nonpregnant women, 20 to 70 years of age, reported that oral administration of nattokinase produced a who met the enrollment criteria and gave written informed gradual enhancement of fibrinolytic activity in the plasma, consent, were assigned to one of the 3 groups: the Healthy as indicated by assays of euglobulin fibrinolytic activity and Group, the Cardiovascular Group, and the Dialysis Group, the production of tissue plasminogen activator [6]. The according to their conditions, aggregating 15 in each group. potent fibrinolytic activity of nattokinase suggests that it Study participants underwent screening evaluations up to 30 may be a nutraceutical for improving circulation and days before administration of the investigational nattokinase reducing the risk of cardiovascular disease (CVD). How- product. Eligible for inclusion into the Cardiovascular ever, no evidence is available in the literature on whether a Group were patients with any one type of CVD such as supplement of nattokinase is beneficial for humans with risk coronary artery disease, peripheral arterial occlusive factors of CVD. disease, history of stroke, transient ischemic attack, Fibrinogen is a strong, consistent, and independent risk pulmonary embolism, deep vein thrombosis, and/or who factor related to CVD [7-9] and even as a possible marker for had at least 2 major risk factors of CVD as listed by the CVD [10,11]. Several studies have also indicated that factor Taiwan National Health Insurance guidelines. These risk VII and factor VIII were highly related to the increase in risk factors were hypertension, smoking, diabetes mellitus, atrial of CVD [9,11-14]. Since the coagulation factors named are fibrillation, lipid disorder, overweight, and physical inactiv- highly associated with CVD (hereafter referred to as “CVD- ity. Patients undergoing regular dialysis at the same institute associated coagulation factors”), they were used here as for at least 3 months were eligible for inclusion into the indicators for assessment of the effects of nattokinase on Dialysis Group. The healthy volunteers (Healthy Group) did CVD. In addition, it is well established that increases in total not have any of the risk factors listed above or abnormal cholesterol (TC), triacylglycerol (TG), and low-density renal function as defined by serum creatinine above 1.4 mg/ lipoprotein cholesterol (LDL-C) concentrations as well as a dL for males or above 1.3 mg/dL for females. Subjects with decrease in high-density lipoprotein cholesterol (HDL-C) known allergies to the component products of the study, concentration are important risk factors for CVD [15-17].A those currently using warfarin (an anticoagulant medication study has shown that natto extracts could lower the levels of for thrombosis or embolism), those with active disease TG and TC in cholesterol-fed rats [18]. In this study, we status, or those suffering from acute diseases were excluded hypothesized that nattokinase could reduce certain factors of from this study. blood clotting and lipids that are associated with an increase 2.2. Investigational product risk for CVD. To test this hypothesis, we investigated the effects of nattokinase on plasma levels of blood coagulation The study product of enteric-coated nattokinase capsule factors and blood lipids in human subjects. We hypothesized contained nattokinase 400 mg (2000 FU per capsule) and that, by administration of nattokinase, the CVD high risk gelatin 100 mg. The suggested usage was 2 capsules once a group should be affected more on the indicators examined day taken at about 30 minutes after dinner. than healthy volunteers because this group might have more 2.3. Efficacy assessment abnormal levels of CVD-associated coagulation factors and blood lipids. Therefore, we recruited 3 groups of subjects, After a 2- to 4-week screening period, subjects were including healthy volunteers, patients with CVD and/or more provided an 8-week supply of the study product to evaluate than 2 of CVD risk factors, and patients undergoing dialysis. the effects of oral intake of the nattokinase capsules. Since Patients undergoing dialysis were included because it is well- most of the assessments made in the study are known to known that CVD is the major cause of complication and fluctuate easily intra- and interindividually, subjects were mortality in these patients [19,20]. Furthermore, although requested to return for evaluations at the same time of day natto has been consumed as a natural food supplement for starting from the screening visit. All laboratory samples were years without safety concerns, an increase in uric acid collected after an overnight fast from all subjects. At every concentration after soybean consumption had been reported study visit, patients were asked to return all of the unused 192 C.-H. Hsia et al. / Nutrition
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