Advances in Health Sciences Research, volume 26 2nd Bakti Tunas Husada-Health Science International Conference (BTH-HSIC 2019) Ethanol Extract Activity of Yellow Pumpkin Flesh (Cucurbita Moschata Duch) on the Cataract Formation Tita Nofianti*), Nurlaili Dwi Hidayati, Vera Nurviana, Ruswanto, Rani Yunda L Department of Pharmacy, Bakti Tunas Husada Health Science College, Tasikmalaya Tasikmalaya, Indonesia *[email protected] contain some minerals, such as calcium, phosphorus, iron, and Abstract—Objectives Oxidative stress is suspected as a major factor vitamins, vitamins B and C, as well as fiber. flesh of the fruit triggering cataracts. Yellow pumpkin contains beta-carotene which is yellow or orange is a sign that it has a very high compounds can reducing oxidative stress, and it is suspected to be carotenoid content [3]. able to reduce the occurrence of cataracts. This study aims at determining and proving the benefits of yellow pumpkin (Cucurbita Indonesian society has long used flesh of yellow pumpkin moschata Duch) as a deterrent in cataract formation in male albino as everyday dishes, and it is also used as a traditional medicine rats (Wistar strain) induced by Sodium Selenite. Macroscopic and to treat various diseases. It has triggered many researchers microscopic features of cataract formation on negative controls conducting research on yellow pumpkin plants ranging from included in mature cataracts (stadium 3), at the dose I and dose III the chemical content within it to the advantage that can be including incipient cataracts (stadium 2) and dose II including obtained from it. Research on the use of yellow pumpkin has immature cataracts (stadium 1). The average parameter diameter of been investigated as antidiabetic, hepatoprotector, anti-cancer, the lens (μm) of group dose II (1738 ± 583) and dose III (1825 ± anti-obesity [4]. In addition, yellow pumpkin is useful as anti- 106) experiences a widening of the lens diameter compared to the HIV, anti-diarrhea, anti-bacterial, laxative, carminative, anti- negative control (1492 ± 52). Whereas, at the dose I (1392 ± 38), there was a narrowing compared to the negative control. The pyretic, anti-oxidant, anthelmintic, and anti-tuberculosis [5]. parameters of lens epithelial cell thickness (μm) at dose I (15 ± According to Valenuela et al. (2011), yellow pumpkin is used 3.46), dose II (15.67 ± 5.03), and dose III (9.67 ± 4.62) of ethanol for the treatment of cataracts. extract of yellow pumpkin showed an increase in thickness lens This study aims at determining the effect of ethanol extract epithelial cells of mouse compared to negative controls (7 ± 1). The of yellow pumpkin fruit in improving the health of eye ethanol extract of the flesh of yellow pumpkin showed the experiencing cataracts from a laboratory rat having been improvement of eye health of albino rats (Wistar strain) induced of sodium selenite. Observations was macroscopically experienced cataract, although it has not been able to improve it on and microscopically done on the rat eyes. the whole. Keywords: cataract, Cucurbita moschata Duch, sodium selenit II. MATERIAL AND METHOD I. INTRODUCTION A cataract is a clouding of the lens in the eye that can A. Procedure reduces the amount of light entering the eye that affects vision. Tools and materials a cataract remaining in the eye for a very long time become The tools used are rotary evaporator, Olympus binocular cause of visual impairment and blindness around the world microscope, paraffin block molding place, tissue processor, making at least 50% of blindness in most developing countries. embedding tools, microtome. The materials used were 2-3 Blindness is estimated to reach 75 million in 2020. Of these, months old yellow pumpkin fruit obtained from the cataracts are estimated at 35 million. This number is equivalent Cimaragas, Ciamis, West Java, sodium selenit, BNF 10%, a to the current combined total population of Australia, New laboratory rat used is male albino rats (Wistar strain) aged 9 Zealand, Sweden, and Denmark including Indonesia. Thus, the burden of cataracts increases [1]. days. Yellow pumpkin (Cucurbita moschata Duch) is a vegetable plant type, but it also can be used for various types of food, B. Research procedure such as: bread, dodol , chips, kolak , manisan and so on which This research was conducted with the following stages: have a fairly complete nutritional content. Those nutritional Material Collection content of foods are carbohydrates, proteins, fats, glycosides, sitosterol, and carotenoids [2]. In addition, those foods also Copyright © 2020 The Authors. Published by Atlantis Press SARL. This is an open access article distributed under the CC BY-NC 4.0 license -http://creativecommons.org/licenses/by-nc/4.0/. 118 Advances in Health Sciences Research, volume 26 Advances in Health Sciences Research, volume 26 The material used is Yellow Pumpkin (Cucurbita moschata Durch) obtained from plantations in Cimaragas, Ciamis, West Test Material Preparation Java. Dose Determination of Sodium Selenite Yellow Pumpkin Plant Determination Based on previous research, sodium selenite dose of 25 Determination of Yellow Pumpkin Plant (Cucurbita moschata µmol/Kg of body weight rats can induce cataracts. Sodium Durch) at Laboratory of Plant Taxonomy, Department of selenite dose is 4,32345 x 10-3 grams/Kg of body weight rats. Biology, Faculty of Mathematics and Natural Sciences, Conversion dose = 0.25 x 10-3 g/200 g of body weight rats = Padjadjaran University, Bandung. 0.25 mg / 200 g of body weight rats rats. The concentration of solvent made is 0.25 mg/mL by dissolving 25 mg in aquabides Simplicia Preparation to 100 mL. The flesh of yellow pumpkin as much as 11.6 kilograms is wet sorted by means of the flesh of yellow pumpkin being cleaned Dose Determination Flesh of Yellow Pumpkin and washed, then drained. After being drained, the flesh of Empirical dose flesh of yellow pumpkin for humans is 100 g yellow pumpkin is cut into small sizes and then dried by of fresh fruit flesh. Dose conversion of yellow pumpkin placing it in an open place with good air circulation and not powder = 100 g/8000 gx 477.6 = 5.97 grams. The dose exposed to direct sunlight with a black flannel cloth covered. conversion of extract = 5.97 g / 200 g x 84.63 g = 2.5262 g. Simplicia is mashed using a blender to make the process of By using a conversion factor of 0.018, a dose of 0.0454 withdrawal of secondary metabolites by solvents easier. gram/200 gram of weight of rats was obtained, 3 variations of dose were made, namely 0.0227 g / 200 gram of weight of Extraction rats, 0.0454 g / 200 gram of body weight rats, and 0.0908 g / The flesh of yellow pumpkin which has become powder 200 gram of body weight rats. extracted by maceration, by soaking the powder of flesh of yellow pumpkin in 70% ethanol for 3x24 hours, every 24 Dose Determination of Ascorbic Acids hours replaced by a new solvent and stirring occasionally. The empirical dose of ascorbic acid of humans is 50 mg. The After 3x24 hours, it is filtered with a filter cloth until it is dose in rat is = 0.05 g x 0.018 g = 0.0009 g / 200 g of body perfectly filtered which is marked with the ethanol color weight rats. The concentration of solvent made is 0.9 mg/mL becoming clear again. After that, the remaining ethanol by dissolving 90 mg in aquadest to 100 mL. solvent was evaporated with a rotary evaporator so that a thick extract of flesh of yellow pumpkin was obtained. Process of Cataract Testing The procedure for cataract testing can be seen in the chart Phytochemical Test below: Phytochemical test conducted includes alkaloid compounds, A laboratory rat aged 9 days were grouped into 6 groups. Each flavonoids, polyphenols, tannins, monoterpenoids and group was induced (except the normal group) with sesquiterpenoids, saponins, quinones, terpenoids and steroids. subcutaneous sodium selenite. 24 hours after being induced, then it was given the test compound. The dose was given Animal Preparation orally to each laboratory rat until it opened its eyes for the first In this study, the animals used male albino rats (Wistar strain) time (± 18 days). Macroscopic observations of all laboratory aged 9 days, and it was placed in a cage at room temperature rats were performed. All laboratory rats were sacrificed at the (25 ± 100C) and relative humidity (55 ± 5%). Then, the end of the study, and the eye organs were taken to make the animals are grouped into 6 groups, in which in each group lens histopathological preparations. The histopathological consisted of 5 animals, and they were given treatment as features of the lens experiencing cataracts in each eye of the shown in table 1: laboratory rats. Table 1. Animal Preparation Setup the histopathology preparations [6,7]. Test Group Treatment Soaking the tissue in 10% of buffer neutral formalin (BNF) Normal PGA 1% (2 mL/200 g of body weight rats) Soaking the tissue in 10% of buffer neutral formalin (BNF) Negative Na. Selenite + PGA 1% (2 mL/200 g of body functioned as a preservative to prevent tissue digestion by weight rats) enzymes or bacteria and to protect the physical structure of Positive Na. Selenite + Vitamin C (0,0009 g/200 g of cells. The process of tissue fixation by using 10% of BNF body weight rats) soaked for 3 days. Dose I Na. Selenite + Ethanol Extract Flesh of Yellow Pumpkin (0,0227 g/200 g of body weight rats) Process of Histopathology Preparation Dose II Na. Selenite + Ethanol Extract Flesh of Yellow It Included organ washing, dehydration process, purification Pumpkin (0,0454 g/200 g of body weight rats) process, vacuum, printing paraffin blocks, cutting tissue Dose III Na.