The Effective Dose and Pattern of Soybean Extract Administration to Regulate Body Weight of Laboratory Rats

Background: Protein in soybean, β conglycinin is responsible for anti-obesity effects by suppressing appetite via stimulation of Cholecystokinin. Cholecystokinin is a hormone released in the digestive tract in response to the intake of protein and suppress appetite for short-term. Detam 1 variety is a high-quality soybean according to the Minister of Agriculture of Indonesia. Soybean protein extract Detam 1 by Deak method contains high levels of β conglycinin. The purpose of this study was to determine the effective dose of protein extract Detam 1 soybean Deak Method (PEDSDM) in reducing food intake, regulate body weight, and plasma CCK level for 14 and 28 days at various dosage and pattern of treatment on male Wistar rats. Methods: There were eleven groups of treatment (n = 3), administrated with extracts at 5 mg/1x/day, 10 mg/1x/day, 20 mg/1x/day, 2.5 mg/2x/day, 5 mg/2x/day, 10 mg/2x/day and 1.7mg/3x/day, 3.4mg/3x/day, 6.7 mg/3x/day, negative control group (distilled water) and positive control group (Sibutramine). Food intake (g), weight loss (g) and measurement of plasma Cholecystokinin levels by ELISA (ng /ml) Results: The results showed that the highest percentage decrease in food intake is: group 3.4mg /3x/ day (p <0.05), inhibition weight gain for 14 days: group 10 mg /1x/ day, for 28 days: group 1.7 mg/3x/day (p <0.05), increased plasma Cholecystokinin levels: group 20 mg /1x/day (p <0.05). Conclusions: The effective dose and pattern administrating the rats for 14 days is extract of 10 mg once a day in the morning, for 28 days is 1.7 mg three times a day. (Health Science Journal of Indonesia 2016;7:17-26)

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Statistical Analysis
Data of food intake, body weight and CCK were analysed XVLQJ $129$ WHVW IROORZHG E\ 'XQFDQ +6' WHVW Ethical approval from Ethical Review Board of Faculty of Medicine, Maranatha Christian University ZLWK 1R .

RESULTS
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As seen in Figure 1, the result of Negative control group along 28 days of treatment, the food intake was EHWZHHQ J GD\ J GD\ EDVHOLQH DQG HQGOLQH 21.5 with p>0.05. Group B, the least food intake was J GD\ WKH KLJKHVW ZDV J GD\ DQG GHFUHDVHG DIWHU 14 days, baseline 23.25 and endline 16.5 with p>0.05. *URXS ' IRRG LQWDNH ZDV EHWZHHQ J GD\ J GD\ and decreased after 7 days, baseline 21.33 and endline S! 7KH KLJKHVW IRRG LQWDNH RI JURXS * ZDV J GD\ ZKLOH WKH OHDVW IRRG LQWDNH ZDV J GD\ LW was very low and tend to decrease after 7 days, baseline 24.00 and endline 17.0, p>0.05. Group K was between J GD\ J GD\ DQG GHFUHDVHG DIWHU GD\V EDVHOLQH 20.33 and endline 17.33 with p>0.05.
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were not in the same column with group A, negative control, so it conclude that the research methods used are considered valid. Most of the treatment group were not in the same column with group A, this suggests that the group treatment had different effects with Negative control, and was assumed that the treatment groups had potencies to control weight JDLQ 7UHDWPHQW *URXS ( + , ) DQG % ZHUH LQ WKH same column with the comparison group K, indicated that this treatment groups had a similar effect with VLEXWUDPLQH DIWHU GD\V RI WUHDWPHQW 7DEOH Jean et al, stated that daily food intake will decrease at condition of high intake of protein diet. 14 High intake of protein was proved can suppressed appetite and followed by decreased total food intake.
A metaanalysis study about energy density reports that the highest effect to satiety, is protein, second is carbohydrate then followed with fat. 15 Hydrolysate protein that comes from soybean showed the best result in inducing CCK secretion. 16 CCK will suppress hunger and short time appetite, so that animal study will lost hunger and appetite for a mean time after being given PEDSDM.
In herbal medicine, higher dosage did not cause better HIIHFW 7KLV FRQGLWLRQ RIWHQ RFFXUHG WKH UHVSRQVH GLG QRW HTXDO ZLWK DVVXPSWLRQ GRVDJH DQG SUHGLFWHG effects. In herbs, besides main active substance, there were another substances, although in a small DPRXQW FRXOG LQÀXHQFH WKHLU SRWHQF\ RU HIIHFW In PEDSDM, besides the main active substances, FRQJO\FLQLQ VXE XQLW WKHUH ZHUH DQRWKHU VXE XQLW OLNH FRQJO\FLQLQ VXE XQLW . DQG . ¶ *O\FLQLQ IUDFWLRQ ZKLFK KDV SRWHQWLDO WR LQÀXHQFH WKHLU satiety effect. In low dose, these substances still in VPDOO DPRXQW DQG QRW \HW LQÀXHQFH WKH HIIHFWV ,Q therapeutic dose, these byproduct substances still not caused any effects, but in higher dose, it will begin ruin the effects of satiety so if we raise the dosage, we can't expect better result. In this study, the phenomena that happened was inhibition of weight gain and after the percentage result of average body weight changes were calculated, among treatment groups showed highly VLJQL¿FDQW GLIIHUHQFH RQ GD\V RI WUHDWPHQWV S 0,018), 28 days of treatments (p < 0,001). If we compared the body weight graphic between NC group and treatments group, there was a difference RI VORSH . WDQJHQVLDO WKDW WKH JUDSKLF RI WUHDWPHQW groups had a smaller angle (graph more sloping) which indicates an increase in body weight that occurs is smaller than the negative control group. Effect of PEDSDM that happens in this study is inhibit the increasing weight gain or controlled body weight in male Wistar rats.
In once daily treatment, the group that showed the OHDVW UHVXOW ZDV JURXS ( GRVDJH PJ UDW RQFH GDLO\ food intake of 3 rats in group E was 4g, 7g and 7g ZLWK &&. OHYHO QJ PO QJ PO DQG QJ PO DYHUDJH QJ PO 3('60 GRVH PJ UDW GD\ VKRZHG JRRG HIIHFWV WR FRQWURO ERG\ ZHLJKW in 14 days, and caused high level of CCK plasma mean the inhibition of weight gain was caused by CCK that suppress appetite. In this study, the once daily treatment were given at 08.00 a.m so it can be concluded that administration pattern once daily is good and also means CCK were good given not in divided dose and best given in the morning (08.00 D P 7KLV UHVXOW ZDV DSSURSULDWH ZLWK 0HULQR ¶V study, which stated that CCK primary effects is in the morning, assumed that circadian had effects towards CCK. Aoyama study stated that rats which administered with soybean protein isolated for 14 days showed dramatically decrease fat and body weight compared with rats which given diet QJ PO DQG VLJQL¿HV its potential better in increasing plasma CCK levels. 7KLV LV EHFDXVH VLEXWUDPLQH LV QRW LQFOXGH SURWHLQ RU fat, which has the ability to trigger the secretion of CCK. In this study, treatment of PEDSDM dosage of 20 PJ UDW RQFH D GD\ FDXVHG WKH KLJKHVW OHYHOV RI &&. SODVPD QJ PO 7DEOH DQG )LJXUH EXW showed only good for weight control on 14 days treatment, while treatment for 28 days just showed PRGHUDWH HIIHFW 7KLV VKRZHG WKDW WKH &&. PDLQO\ plays a role in weight loss for the treatment of short time period (14 days) and better administered in a single dosage (not divided pattern) in the morning. Effect of CCK lowered portions (meal size) in rats that ate as much as they like (feed DG OLELWXP) is better than the size of overall food in fasted rats. 23,24 However CCK in small level have already shown effect, referring to another study that says that the PLQLPXP OHYHO RI &&. HIIHFW ZDV QJ UDW HYHQ RQ lower dosage of CCK can show effects in experimental animals are fasted or at the time of the morning, it is estimated that circadian factors also affect the effect of CCK. 20,24 In rats that fed DG OLELWXP given treatment GRVDJH &&.