Malnutrition is caused by inadequate protein intake and affects growth factor. High protein from patin (
Malnutrition is generally caused by restricted dietary intakes and ended with poor linear growth even in early life (
Malnutrition is caused by a lack of nutrients intake, such as a protein restriction diet (
IGF-1 and IGFBP-3 are produced in the liver. They have a function in mediating the protein metabolism and controlling the growth factors which are accumulated in somatic cells (
Fish protein concentrate is a product resulting from the relieving moisture and fat content. Bioactive compounds and peptide contained in fish have benefits to control growth (
There are various doses of patin fish protein concentrate that affect insulin-like growth factor (IGF)-1 and IGF binding protein (IGFBP)-3 levels of Sprague Dawley neonate rats-induced malnutrition.
Patin fish were bought at fishpond in the Ambarawa subdistrict, Semarang Regency, Central Java Province, Indonesia. The selected patin fish as those that have a good condition as shown by fresh patin that has just been captured, weighs about 1 kilogram for one fish, has fresh flesh, has gills that are still red, the color does not change and odorless. Then the patin fish were filleted and cleaned. The filleted fish were ground using a food processor with the addition of salt (0.5%) and NaHCO3 (1.5%). Next, fish pasta was steamed for 30 minutes. The steamed pasta was then pressed and extracted by 96% food-grade ethanol with a 3:1 patin ratio. The extraction was carried out twice to fully relieve the moisture and fat content. The extracted fish was then dried at a temperature of 40 °C and mixed until refined size (60 mesh). The PFPC was wrapped in foodgrade silica gel and stored in the bottle at 4 °C (
The proximate analysis includes moisture, protein, ash, fat, and carbohydrate content (by difference) (
This research was a true-experiment study with a randomized pre-post test and has a control group design. The animal used was male Sprague Dawley rats, aged 21 days induced with a low 8% protein diet (L8PD), except health control. The acclimatization was held in the laboratory of Gadjah Mada University, Yogyakarta. The rats were placed in each group and numbered. They were placed at a regulated temperature (21°C) and a clean cage. They were given ad-libitum water during the experiment. Animal care in the laboratory was carried out in accordance with the animal Laboratory Guideline from the Central Laboratory for Food and Nutrition Studies, Gadjah Mada University, Yogyakarta.
Thirty rats were divided into five groups, namely groups of normal control (K1), malnutrition control (K2), malnutrition with PFPC 13.26 mg.g-1 BW/day (X1), malnutrition with PFPC 19.89 mg.g-1 BW/d (X2), and malnutrition with casein supplement 13.26 mg.g-1 BW/d (X3) (
Ingredient compositions (g/kg) of low 8% protein diet (L8PD) fed to malnutritiomn-induced rats.
Ingredients | L8PD (g)* |
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|
|
Casein purified high nitrogen | 80 |
Corn starch | 780 |
Cottonseed Oil | 100 |
Salt mixture U.S.P XIV | 40 |
Total | 1000 |
Note: * L8PD for K2,X1,X2, and X3.
This study was approved by the Health Research Ethics Commission of the Faculty of Medicine Diponegoro University Semarang through ethical clearance No. 131/EC/H/KEPK/FK-UNDIP/X/2019.
Results were expressed as mean ± SD (for normally distributed data), otherwise, it was expressed as median (min-max). Statistical difference was analyzed by using a one-way analysis of variance (ANOVA) followed by post hoc Bonferroni for normally distributed data, otherwise, the Kruskal-Wallis test followed by Mann-Whitney-U-test was used. The software used was SPSS software version 22 (SPSS Inc, Chicago, IL, USA). Spearman’s correlative test was used to analyze the relationship between variables. Statistical analyses were done by the computer. The differences and correlations were considered significant at
Related data about PFPC was shown as proximate analysis (Table
Proximate analysis of PFPC.
Nutrient Content | % |
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|
|
Moisture | 7.24 ±0.35 |
Ash | 2.77 ±0.07 |
Protein | 81.07 ±0.56 |
Fat | 4.08 ±0.18 |
Carbohydrate | 4.83 ±0.30 |
Note: * L8PD for K2,X1,X2, and X3.
Wilcoxon test carried out on the body weight before and after the intervention, showed that the intervention of PFPC increased the body weight in the all-treatment-group as shown in Table
Statistical analysis of body weigth, IGF-1 and IGFBP-3 (control groups).
Variables | K1 | K2 | ||||||
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Pre | Post |
|
Δ | Pre | Post |
|
Δ | |
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BW | 106.5 (104-112) | 152.5 (149-158) | 0.026 | 48 (38-53) | 75.50 (70-81) | 98.5 (92-101) | 0.026 | 22.00 (20.00-23,00) |
IGF-1 | 41.32 (39.77-42.53) | 42.35 (40.81-43.22) | 0.026 | 1.03 (0.69-1.38) | 69.04 (67.67-71.11) | 70.59 (68.70-72.14) | 0.027 | 1.20 (0.69-1.72) |
IGFBP-3 | 40.89 ±2.69 | 42.07 ±2.43 | 0.004 | 1.17 ±0.57 | 92.16 ±1.96 | 94.53 ±2.23 | 0.034 | 2.37 ±1.99 |
Note: a
Statistical analysis of body weigth, IGF-1 and IGFBP-3 (group with PFPC and casein intervention).
Variables | P1 | P2 | P3 |
|
|
|
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Pre | Post |
|
Δ | Pre | Post |
|
Δ | Pre | Post |
|
Δ | ||||
|
|||||||||||||||
BW | 77 (71-80) | 108 (100-109) | 0.027 | 29.5 (28-34) | 77 (73-82) | 110 (107-118) | 0.027 | 34.50 (33-36) | 76.5 (75-79) | 107 (104-109) | 0.026 | 29.5 (29-32) | 0.005 | 0.001 | 0.001 |
IGF-1 | 69.56 (67.67 - 99.35) | 53.72 (52.51 -54.92) | 0.028 | -16.01 (-44.42 - 13.77) | 68.70 (66.29 -71.11) | 39.77 (34.95 -41.15) | 0.028 | -29.61 (-33.75- -25.14) | 68.53 (66.63 -69.73) | 44.42 (42.87 -45.63) | 0.027 | -23.59 (-25.83 - -23.07) | 0.003 | 0.001 | 0.001 |
IGFBP-3 | 92.56 ±2.43 | 66.23 ±2.37 | 0.001 | -26.32 ±3.23 | 90.59 ±2.65 | 49.33 ±1.23 | 0.001 | -41.26 ±2.79 | 92.71 ±4.50 | 46.56 ±1.45 | 0.001 | -36.15 ±3.60 | 0.001 | 0.001 | 0.001 |
Note: a
Patin Fish Protein Consentrate.
Changes of body weight of neonate rats during the study. Malnutrition was induced from day 0 to 21st by giving low 8% protein diet (L8PD). PFPC intervention was started from day 24th to 38th. Three days acclimatization was done from day 21st to 24th.
The IGF-1 levels decrease at the end of the intervention, and significantly different (
The IGFBP-3 levels experienced a decrease after the performance of the intervention. The levels were significantly different (X1, X2 and X3 had the same
The Spearman test on all data from all rats at the end of the study showed there was a very strong correlation that was found between the variables (IGF-1 and IGFBP-3). A very strong positive correlation was observed between IGF- 1 and IGFBP-3 (
Malnutrition can be ended with a linear growth disruption. Restriction of protein intake during the lactation period affects the disruption of long term linear growth (
Inflammation is a possible indication in the intervention group which is shown through the decreasing serum IGF-1 levels in neonatal rats. Indications of inflammation are thought to occur due to the placement of each group of neonate rats in one cage. The same result was also found in other previous studies showing the effect of interleukin 6 (IL-6) inflammatory biomarkers on IGF-1 descent. The decrease of IGF-1 is related to the mitogenic function of IGF-1 thereby inducing a decrease of its concentration in cell proliferation (
These indications of inflammation due to chronic malnutrition are also associated with GH resistance response, which modulates gluconeogenesis (
Other previous studies have assessed that serum IGF-1 levels can be assessed at four weeks of intervention. Meanwhile, this study assessed the IGF-1 in two weeks’ intervention. This is also supported by the insignificant assessment of protein synthesis which ended with a disruption of metabolism especially the disruption of IGF-1 synthesis (
Decreasing serum IGFBP-3 levels can cause disrupt IGF- 1 modulation. IGFBP-3 is the principal binding protein that modulates the action of IGF-1 as a growth factor. Disturbances that are caused by protein restriction and are not properly repaired realized that the pituitary-GH-IGF axis binding protein has a reciprocal effect (
Proteolysis is the most possible mechanism in this study which has a function to reduce serum IGFBP-3 levels. Both levels of IGF-1 and IGFBP-3 which were decreased show that the mechanism most likely caused by proteolysis (
Interestingly, other growth factors besides IGF-1 and IGFPB-3 might be involved and affected the increasing body weight of neonate rats. These growth factors show the effect of suppression on IGF-1 and IGFBP-3 but support the mechanism of other growth biomarkers which is associated with the increasing body weight (
Malnutrition condition, which was caused by cachexia symptoms and inflicted from malignancy response, also observed a similar indication. It was shown significantly by a specific combination of high-protein intake intervention which contained leucine (
The findings in protein intervention also improve clinical symptoms due to protein restriction. Other growth factors can affect the increase in body weight of malnutrition with re-feeding (
The increase of body weight which is indicated by increased GH is also associated with carbohydrate and fat homeostasis (
Furthermore, amino acid, especially lysine and leucine which are high in PFPC products could also change weight gain. We found in this study that the PFPC contained protein amounting 80% (Table
The high protein concentration also supports the lipid and carbohydrate pool of neonate rats. Increased protein intake can affect the amount of lipids and carbohydrates stored in adipose tissue, especially white adipose tissue (WAT) (
Limitations of this study we do not examine other growth indicators on this intervention except the body weight. Discussing another growth factor, we also do not examine another related growth factor that can relate to and support the hypothesis of this study. Inflammation is not measured in this study, as expected to exist in chronic malnutrition neonate rats. The proteolytic, which can be the most possible involved mechanism in this study is also not concerned. We did not concern about the digestible amino acid of PFPC which can be compared to other products in case of body weight gain of malnutrition rats. Other biomarkers that relate to lipid metabolism also not a concern.
The administration of patin fish protein concentrate with various doses tested, significantly increased the body weight, but decreased IGF-1 and IGFBP-3 serum level of malnutrition-induced neonatal rats. The administration of patin fish protein concentrate with dose 13,26 mg.g-1 body weight per day is the most effective dose in increasing body weight of malnutrition-induced neonatal rats.
This research did not receive any specific grant from funding agencies in the public, commercial, or not-forprofit sectors.