PRELIMINARY:

The rapid progress of food science and technology and pharmaceuticals has provided scientific evidence that some types of Indonesian food are believed to be useful for improving health and treatment, one of which is functional food products that can be used as a treatment for diabetes mellitus.

Antidiabetic drugs may be needed for people with diabetes who are allergic to insulin or who do not use insulin injections. Excessive use of antidiabetic drugs can cause unwanted side effects, so that experts develop a safer antidiabetic traditional treatment system because traditional drugs contain safe antioxidants without causing side effects.

Antioxidants are chemical compounds that can contribute one or more electrons to free radicals so that free radicals can be quenched. Based on the source of the acquisition¹, antioxidants are divided into two: natural and artificial antioxidants (synthetic). The human body does not have antioxidant reserves in excess amounts, so if there is excessive exposure to radicals, the body will need exogenous antioxidants. Plants that can be used as natural antioxidants are cassava and soybeans.

Banggai sweet potato (Dioscorea alata L.) contains diosgenin, dioscorin, and phenol bioactive compounds that function as antioxidants.² Sweet potato also contains Water-Soluble Polysaccharides (PLA) and inulin which are useful as prebiotics, control blood glucose levels and show an inhibitory effect on blood glucose absorption in blood glucose response tests.³. Soybean (Glycine max (L.) Merr.) also has the potential to reduce blood glucose levels in diabetics. This is due to the presence of isoflavone phytoestrogens (genistein, daidzein, glycitein) which affect glycemic control, insulin sensitivity, dyslipidemia, and kidney function.⁴ Soybean is a food ingredient that is often consumed by the community in various processed forms, one of which is soyghurt⁵

Soy-yamghurt is a fermented product of soy and sweet potato juice made from the addition of lactic acid bacteria and is thought to have antioxidant abilities, function to repair pancreatic cells, and reduce blood glucose levels.

Based on this background, the authors are interested in researching soy-yamghurt formulated with Banggai sweet potato extract and soybean extract with various comparisons between Banggai sweet potato extract and soybean juice whether soy-yamghurt has potential as antidiabetic in male white rats in vivo induced by streptozotocin.

MATERIALS AND HOW TO WORK:

Ingredients:

The materials used in this study included sterile aqua dest, Lactic Acid Bacteria (LAB), namely Streptococcus thermophiles, Lactobacillus bulgaricus, L. acidophilus bifidobacterium (biokul plain) glibenclamide, sugar, gum arabic, solution of NaHCO3 (sodium bicarbonate), Na CMC, soybeans which from Bulan Village, Ampana and proud tubers from Luwuk Banggai, aqua dest, streptozotocin, skim milk.

Phytochemical screening test:

A phytochemical screening test was carried out to detect the chemical composition of plants based on their groups. Used as initial information to determine the class of chemical compounds that have biological activity from a plant. Phytochemical screening tests were carried out on chemical compounds including tests for alkaloids, flavonoids, saponins, tannins.

Making yogurt starter:

16grams of skim milk is added with 3% sugar, then dissolved in 100ml of hot water while stirring well and then the temperature is lowered to 45ºC. Adding a commercial yogurt starter culture (plain biokul) as much as 5% of the volume of the mixture and then stirred. Then it is closed using polyethylene plastic that has been perforated using a needle. The resulting mixture was incubated at a temperature of 40-45ºC. Then store the yogurt starter that has been produced in the refrigerator at 4ºC and under anaerobic conditions⁶.

Producing sweet potato juice and soybean juice:

The extracted sweet potato juice was cleaned by peeling the tubers of the proud yam. Furthermore, washing is carried out to remove the remaining dirt on the tuber flesh, after washing it is then refined with a ratio of 1:3 w/v (Herlina et al., 2018) then filtered using a filter cloth that has been blanched and then the juice is taken.⁶ Soybeans were sorted and washed thoroughly and then boiled 2 times for 30 minutes, before and after soaking the 0.2% NaHCO3 solution for 30 minutes. Then the soybean skin is separated by kneading and washing it many times until it is clean by adding hot water (100º) to the skinless soybeans and then grinding or blending. The resulting soybean porridge is filtered using a filter cloth that has been blanched to obtain soybean juice and then left on low heat at 80ºC for 20 minutes.⁶

Making Soy-yamgurt:

Soy-yamgurt is made from cassava extract and 79.4% soybean extract in a ratio of 1:1, 1:2, and 2:1. Then 15% skim milk, 2% sugar, 3% starter and 0.6% NaCMC mixture were added. Then do the incubation process at a temperature of 40ºC for 6 hours. Soy yamgurt that has been formed is then stored in the refrigerator at 4ºC. Anaerobic conditions⁶

Testing of antidiabetic activity in test animals:

A total of 30 were divided into 5 groups, each group divided into 6 test animals with each group, namely Normal control (NaCMC), negative control (NaCMC + STZ), positive control (glibenclamide + STZ), formula 1(1:1) given 3.4ml, formula 2(1:2) was given 3.4ml and formula 3(2:1) administration of 3.4ml. Before the test animals were used, the test animals have fasted for 16 hours, and then blood glucose levels were checked using a glucometer. After that, the initial blood glucose level was measured (on day 0). Then streptozotocin was induced at a dose of 40mg/kg BW intraperitoneally except for the normal group. On the 7th day, the rats fasted for 16 hours, and then the second blood glucose level was measured to determine blood sugar levels after streptozotocin induction. After the fasting blood glucose levels of the mice reached a state of hyperglycemia, the mice that had become hyperglycemic were treated according to their group for 28 days. Blood glucose levels were measured every 1 week after treatment. The measurement data of blood glucose levels before and after the treatment obtained were recorded and analyzed

Data analysis:

The results of the measurement of blood glucose levels obtained were taken using the Kolmogorov Smirnov test first to determine the normality value. After that, it was continued with the lavender test to determine homogeneity. If the data shows a normal and homogeneous distribution, then proceed with parametric analysis using the one-way ANOVA test and continue with the LSD further test. If the data is not normally distributed, Kruskal Wali's nonparametric analysis and Mann Whitney further test are performed.

RESULTS:

Phytochemical Screening Test Results

Table 1 results of the phytochemical test of the extract of Banggai sweet potato (Dioscorea alata L)

S. No	Secondary Metabolic Compound	Reactor	Results
S. No	Secondary Metabolic Compound	Reactor	Red castor leaf ethanol extract
1.	Flavonoids	5ml Hydrochloric acid 2n + 3 drops dragendrof (brown-black precipitate)	Formation of orange color
2.	Alkaloids	10ml hot water+shake+hydrochloric acid 2N (forms 1-10cm foam)	A dark brown precipitate formed
3.	Saponins	20ml hot water + 3 drops of 10% NaCl + FeCl3 (Formed black blue or greenish-black)	Foam is formed
4.	Tannins	10ml hot aqua dest + 3 drops FeCl3 (a bluish-green color is formed)	Formed a bluish-greenish black color

Table 2 Phytochemical test results of the extract of Banggai sweet potato (Dioscorea alata L)

No	Secondary Metabolic Compound	Reactor	Results
No	Secondary Metabolic Compound	Reactor	Red castor leaf ethanol extract
1.	Flavonoids	5ml Hydrochloric acid 2n + 3 drops dragendrof (brown-black precipitate)	Formation of orange color
2.	Saponins	20ml hot water + 3 drops of 10% Nacl + FeCl3 (Formed black blue or greenish black)	Foam is formed

Table 3 Average Blood Glucose Level

days to	0	7	14	21	28
N	99.2±8.927	99.2±8.927	105.8±5.119	101.0±8.185	100.4±7.635
K	87.0±7.906	291.0±134,625	280.6±65.359	278.2±71.482	298.8±13,971
K+	94.4±3.975	250.4±23.522	161.4±72.851	141.0±53,558	81.2±7.396
F1	90.8±12.458	326.8±78.643	313.2±123,837	196.6±82.160	178.4±61,484
F2	92.4±7.436	347.6±123,470	170.2±28.039	116.6±19.629	108.0±17.986
F3	92.0±9.192	264.0±109,902	221.4±160.355	172.0±75.561	134.4±15.143

Note: N = normal,K− = negative control, K+ = positive control, F1 = formula 1, F2 = formulas, F3 = formula 3

Figure 1 Profile of blood glucose levels

Note: - the same letter on the same day shows an insignificant difference and different letters on the same day show a significant difference

DISCUSSION:

This study used soybeans (Glycin max (L) Merr) and sweet potato (Dioscorea alata L) which aims to obtain the extract from soybeans and sweet potato as raw materials for making soy-yamghurt formula with various concentrations, namely formula 1(1:1). contains 39.7ml of Banggai sweet potato extract and 39.7ml of soybean extract, formula 2(1:2) contains 26.46ml of Banggai sweet potato juice and 52.92 soybean extract, formula 3(2:1) contains 52.92ml of Banggai sweet potato juice and 26.64ml of soybean juice.

This study used male white rats (Rattus norvegicus) as test animals because these animals tend to be easy to adapt, have easier handling and maintenance, have high reproductive ability, hormonally male rats are more stable. After all, they do not experience estrus and pregnancy phases which can interfere with the final results of a study, production and reproduction characteristics tend to be similar with mammals⁷. Male white rats were divided into 6 treatment groups, each group consisting of 5 rats, Male white rats were adapted for 14 days to adapt to the new environment and prevent stress during treatment. Furthermore, male white rats have fasted for 16 hours to normalize the metabolism in the body of the test animals and to reduce food interaction factors, and speed up gastric emptying time.⁸. Then the induction of streptozotocin at a dose of 40 mg/kg BW was administered intraperitoneally. Streptozotocin acts directly on pancreatic beta cells with its cytotoxic action mediated by reactive oxygen species (ROS) so that it can be used as an induction of diabetes mellitus by triggering an increase in the production of excess free radicals and causing oxidative stress.⁹ Then measured blood glucose levels using a glucometer 5 times on days 0, 7, 14, 21, and 28.

The study began with measuring the initial blood glucose level (day 0) to determine the initial blood sugar level before the study. The results of the glucose measurement can be seen in Table 3 and Figure 1. The statistical results of the one-way Anova test show that there is no significant difference between all treatment groups. This can be seen from the value of P = 0.395 (P>0.05). shows the results of the average blood glucose levels of normal rats. The measurement data on the 7th day can be seen in table 3 and figure 1. seen from the one-way Anova test with p-value = 0.005 (p <0.05) shows that the average blood glucose level of rats has increased except for normal controls. This shows the effect of streptozotocin according to the literature stating that the administration of streptozotocin can trigger an increase in the production of excess free radicals and cause oxidative stress which has a high role in damage to pancreatic cells so that it can increase blood glucose levels.¹⁰

The measurement data on the 14th day can be seen in table 3 and figure 1. Judging from the results of the one-way ANOVA test with a value of p = 0.015 (p < 0.05). showed the results of the treatment group had given effect to lowering blood glucose but had not reached normal values. The measurement data on the 21st day can be seen in table 3 and figure 1. Seen from the one way ANOVA test with p-value = 0.001 (p < 0.05). The results showed that all treatment groups had the effect of reducing blood glucose levels. This is thought to be caused by lactic acid bacteria (LAB) contained in the soy-yamghurt formula, under the statement by¹¹ that the administration of L. casei fermented milk may have the opportunity to lower blood glucose levels.

The data on the 28th day of measurement can be seen in table 3 and figure 1. Judging from the one-way ANOVA test with a value of p = 0.000 (p <0.05). The results showed that formula 2 had the effect of reducing blood glucose levels, where the effect of lowering blood glucose levels as an effective formula that could have an impact on reducing blood glucose levels was almost the same as normal control. The difference in the effect of lowering blood glucose levels between the treatments of formula 1, formula 2, and formula 3 is thought to be due to the formula ingredients contained in each given formula having formula variants, namely formula 1 (1:1), formula 2 (1:2) and formula 3 (2:1). Giving soy-yamghurt fermentation in formula 1, formula 2, and formula 3 can reduce blood glucose levels.

The soy-yamghurt treatment group experienced a decrease in blood glucose levels which was assumed to be due to the activity of antioxidant compounds and oligosaccharides in soy-yamghurt which had a hypoglycemic effect. diosgenin from the Dioscorea group can inhibit -glucosidase and -amylase which are agents that break down starch into more complex carbohydrates such as glucose so that diosgenin can indirectly lower blood glucose levels, Herbal drugs treat diabetes by improving insulin sensitivity, increasing insulin production and/or decreasing the amount of glucose in blood ¹²^,13. Dioscorin is a compound that is included as an alkaloid, an alkaloid compound that has a mechanism for reducing blood glucose levels. Alkaloids also can efficiently stop free radical chain reactions. In reducing blood glucose levels, these compounds can inhibit the absorption of glucose in the intestine and increase glucose transport in the blood¹⁴. Isoflavones are flavonoid compounds that have a significant role as antioxidants that can regenerate damaged pancreatic cells. In addition, flavonoids can delay, slow down, and prevent the lipid oxidation process that plays a role in the MDA production process and improve insulin receptor sensitivity so that insulin deficiency can be overcome and there is an improvement in pancreatic cell tissue ¹⁵, The mechanism of probiotics in the gut increases the production of short-chain fatty acids (SCFA)- acetic, propionic and butyric. SCFAs are an important source of energy and are ligands for the free fatty acid receptors (FFAR) – Gpr41 (FFAR3) – (Gpr41 – G-protein coupled receptor) and Gpr43 (FFR2) in intestinal cells. Because SCFA activates these receptors, it can reduce insulin resistance, increase the proliferation and development of pancreatic cells. SCFAs can stimulate GLP-1 and GLP-2 secretion thereby increasing insulin expression and possibly contributing to increased insulin sensitivity, and decreased low-grade inflammation associated with type 2 diabetes mellitus, Herbal medicines have been main source of primary healthcare in all over the world. From ancient times, plants have been catering as rich source of effective and safe medicines^16,17.

CONCLUSION:

The soy-yamghurt product affects reducing glucose levels with an effective formula for reducing blood glucose levels, namely formula 2 with an average blood glucose level of 108.0 mg/dL.

SUGGESTION:

It is hoped that further tests can be carried out in situ examination of glucose absorption tests with an inverted pouch intestine.

ABBREVIATION:

reactive oxygen species (ROS), free fatty acid receptors (FFAR), acetate, propionic and butyric (SCFA), glucogen like peptide (GLP), lactic acid bacteria (BAL), malondialdehyde (MDA), milligrams (mg), deciliters (dL)

REFERENCE:

1. Tandi J, Handayani TW, Tandebia M, Wijaya JA. Effect of Parkia speciosa Hassk Peels Extract on Total Cholesterol Levels of Hypercholesterolemia Rats. 2020; 14(3):2142-2146.

2. Yuniastuti A, Iswari RS, Susanti R. Antioxidant Acitivity in Various Processed Products of Inferior Local Tubers (Dioscorea sp. L.). 2017; (KnE Life Sciences):35-40.

3. Yofananda O, Estiasih T. Potential of Local Tubers Bioactive Compounds as Blood Glucose Lowering Levels: Literature Review. 2016; 4(1):410-416.

4. Nugraheni K, Bintari SH. Antidyslipidemic activity of tempeh flour and soy milk on lipid profile of streptozotocin-induced diabetic rats. 2016; (18).

5. Rustanti N, Nafsih VZ, Avisha RN, et al. Effect of Yoghurt and Cinnamon Soyghurt (Cinnamomum burmannii) on Blood Glucose Levels, Serum Insulin, and Malondialdehyde in Pre-Metabolic Syndrome Rats.J Nutrition Indonesia. 2019; 8(1):60-68.

6. Kwanariesta J, Rusmarilin H, Suhaidi I. Analysis on Soy-Yamghurt Potential as Antidiabetic in Streptozotocin-Nicotinamide Induced Rats. 2018; 01(01).

7. Azhari D, Yuliet, Khaerati K. Activity Test of White Oyster Mushroom Powder (Pleurotus ostreatus (Jacq.) P. Kumm) Against Blood Glucose Levels in Animal Models of Hypercholesterolemia-Diabetes. Galen J Pharm. 2016; 2(2):96-102.

8. Saud M, Jumain, Stevani H. Effect of ethanol extract of daruju (Achantus iliciafolius L) leaves on blood glucose levels in rabbits (Oryctolagus cuniculus). J Chem Inf Model. 2019; 53(9):1689-1699.

9. Tandi J, Sutrisna INE, Pratiwi M, et al. Potential Test of Nephropathy Sonchus arvensis L. Leaves on Male Rats (Rattus norvegicus) Diabetes Mellitus. 2020; 12(5):1-6.

10. Saktra et.al 2018. 2001, Szkudelski T. The Mechanism of alloxan and streptozotocin action in B cells of therat pancreas. Physio. res. 50(6): 537-546.Bul Vet Udayana. 2018; 10(2):116. doi:10.24843/bulvet.2018.v10.i02.p02

11. Sari PP, Hasan M, Sayuti A. 3. Lactobacillus casei Fermented Milk as a Treatment for Diabetes in Mice (Mus musculus). 3 Lact casei Fermented Milk as a Treat Diabetes Mice (Mus musculus). 2017; 11(1):15-19. doi:10.21157/j.med.vet..v11i1.4088

12. Tinggi S, Pharmacy I, Mas P. Qualitative and quantitative determination of secondary metabolites and antidiabetic potential of Ocimum basilicum L . LEAVES EXTRACT. 2021; 14(1):622-628.

13. M. R. Chitra Devi, B. Ramesh. Hypoglycemic activity of Leaves of Bougainvillea spectabilis extract in Streptozotocin-Induced Diabetic Rats. Asian J. Pharm. Res. 2018; 8(2):99-103

14. Tandi J, Handayani TW, Tumanan IR, Wijaya JA. The effect of Myrmecodea tuberose Jack ethanol extract on streptozotocin-induced diabetic nephropathy rats. 2020; (4626):2279-2284.

15. Tandi J, Danthy R, Kuncoro H. Effect of Ethanol Extract from Purple Eggplant Skin (Solanum melongena L) On Blood Glucose Levels and Pancreatic Cells Regeneration on White Rats Male Hypercholesterolemia-Diabetic. doi:10.5958/0974-360X.2019.00494.3

16. Asgharzadeh F, Tanomand A, Ashoori MR, Asgharzadeh A, Zarghami N. Investigating the effects of Lactobacillus casei on some biochemical parameters in diabetic mice. J Endocrinol Metab Diabetes South Africa. 2017; 22(3):47-50. doi:10.1080/16089677.2017.1378460

17. Shreyash Tripathi, Brijesh Kumar Saroj, Mohd Yaqub Khan. Pharmacological Evaluation of Folk Medicinally used Plant by usin Streptozotocin Induced Rat Models. Asian J. Pharm. Tech. 2018; 8 (4):231-243.

Received on 12.08.2021 Modified on 09.10.2021

Research J. Pharm. and Tech 2022; 15(9):4139-4143.

DOI: 10.52711/0974-360X.2022.00695