The Effect Of Giving Red Dragon Fruit (Hylocereus polyrhizus) Extract On Spermatozo Motility In White Rats (Rattus norvegicus) Wistar Line With High Fat Induction

Hyperlipidemia is a condition when high level of blood lipids and causes morphological changes in spermatozoa. Red dragon fruit (Hylocereus Polyrhizus) is one of the plants as a source of antioxidants and has unsaturated fat content that is needed in the spermatozoa maturation process. The aims of this the study is to determine the benefits of red dragon fruit extract on the motility of white rat spermatozoa (Rattus norvegius) induced by high-fat diet. The study is design by true experimental research and research only testing group design. This research conducted in the Pharmacology and Therapy Laboratory, Padjajaran University Education Hospital, in January 2020 using white rats, male . ± 8 weeks wistar strains weighing ± 200-250. The study sample was divided into several groups such group I as a control, group II as a negative control, group III as positive group with simvastatin dose 0.72 mg / day, group IV as research group 1 using red dragon fruit extract dose of 60 mg / 200 grBB / day for 52 days, and group V as group 2 therapy using dragon fruit extract dose of 60 mg / 200grBB / day for 104 days. The results showed that the mean percentage of motility of rat spermatozoa increased in the relief group with the supplement of red dragon fruit extract by dosing of 60 mg / 200 grBB / day for 104 days, or 51, 2%. This study conclude that red dragon fruit has a superior effectivity better than simvastatin in increasing the motility of spermatozoa.


Introduction
Hyperlipidemia is a condition of increasing blood lipid levels characterized by increased levels of total cholesterol, Low Density Lipoprotein (LDL), and triglycerides in the blood that exceed normal limits 1 . In a study conducted by Harini (2009) 2 it was reported that the normal condition of total cholesterol is amounting to 10 -54 mg / dl. Then in a study conducted by Nugroho (2013) 3 it was reported that the level, which will cause changes in spermatozoa morphology. Feeding the mice a diet supplemented with fat led to an increase in cholesterol levels and a decrease in some gonadal cells. This will affect the motility of the spermatozoa so that the decreased motility of the spermatozoa will affect the fertilization process. In addition, a high-fat diet can cause impaired function of the pituitary-pituitary-gonadal pathway and a disruption of the spermatogenesis process and a decrease in HDL and an increase in total cholesterol will cause male erectile dysfunction 7 . If there are excess free radical compounds in the body, the body will not be able to handle it so the body needs a supply of antioxidants from outside to neutralize the radicals that are formed. To prevent the bad effects of free radicals, antioxidants are needed.
Antioxidants play an important role in protecting spermatozoa against ROS, this may help repair spermatozoa abnormalities due to free radicals so that it will increase the quality and quantity of spermatozoa 1 .
Antioxidants are compounds that can inhibit the fat oxidation process. Antioxidants stabilize free radicals by complementing the lack of electrons that free radicals have and will inhibit the chain reaction from forming free radicals. If the formation of free radicals is inhibited, the motility of spermatozoa in obese men will improve 10 One study shows that the nutritional components of fruits and vegetables can lower cholesterol levels.
Currently there are more and more studies on fruits that have a high antioxidant content, one of which is red dragon fruit 11 Red dragon fruit (Hylocereus Polyrhizus) is a plant that is used as a source of antioxidants. Red dragon fruit is believed to reduce cholesterol levels, balance blood sugar levels, prevent colon cancer, and increase fertility. In addition, red dragon fruit see ds also contain unsaturated fats which are needed in the maturation process of spermatozoa 1 From the research results, red dragon fruit can help maintain the survival of spermatozoa and also plays an important role as a fertility agent because it contains antioxidants and anti-proliferative properties. 12 Along with the times, dragon fruit is not only used as decoration and food but also used as a fruit which is one of the natural herbal medicines. Dragon fruit contains vitamin E which is high enough to increase fertility. Based on the content of substances contained in red dragon fruit, researchers are interested in conducting this research.

Research design
The research conducted was a true experimental research design with a test only control group design.
The purpose of data collection is to determine the effect of the treatment variable (independent variable) on the impact variable (dependent variable) 13

Population and sample
The study population was male white rats. The samples of this research were white rats (Rattus Novergicus), Wistar strain, males weighing ± 200-250 and aged ± 8 weeks.

Research variables
The independent variable in this study was the red dragon fruit variable and the dependent variable in this study was sperm motility

Operational Definition of Research Variables
The operational definition can be described as

Standard Feed
The selection of Bravo-512 feed is based on the ease of access to obtain it, and the similarity in the percentage of nutrient content with other standard diets in experimental rat studies. The standard Bravo-512 feed will be provided ad libitum 17 .

High Fat Feed Making
Feed making is done by mixing 50 grams of egg yolk (5%) and 100 grams of goat fat (10%) in 1000 grams of standard feed. Previously, the goat fat was heated until it melted, then mixed with the beaten egg yolk. This mixture is then put into standard feed, chilled feed, and high fat feed is ready to be served for male rats as a treatment to increase male rat choleterol levels. 20 g / day 18 feed will be given

Preparation of CMC (Carboxymethyl Cellulose) Solution
The solution is made by: Making dragon fruit extract preparations is carried out every time you will brush by calibrating the electric scale first, then placing the measuring cup on the scale, first measuring the weight of the measuring cup.
Then enter the red dragon fruit extract, namely: Red dragon fruit extract = 60 mg x 6 rats = 360 mg = 0.36 gr. Then mix with 2 ml of CMC / mouse solution (2 ml x 6 tail = 12 ml of CMC solution) and stir until homogeneous After the red dragon fruit extract preparation is ready, then prepare a 5 ml stomach sonde and take 2 ml of the solution for each mouse and give it to the mice with certainty and with care.

Preparation of Simvastatin
Making After the simvastatin preparation is ready, take 2 ml of simvastatin solution using a gastric swab to be given to the treatment group I rats for 52 days at 10 am.

Treatment Intervention in Experimental Animals
After the rats were acclimated for 7 days, all rats were divided into 5 groups, where one group contains 6 rats using random sampling with the following weeks.

Surgical Procedure And Sperm Sampling
Termination and quality analysis of spermatozoa

Motility Statistics Test of Mouse Spermatozoa
The results obtained were analyzed with statistical software using the One Way Anova method because the dependent variable and independent variable had a numerical data scale and this study was comparative because it wanted to compare the effect of giving red dragon fruit extract (Hylocereus polyrhizus) on the motility of spermatozoa given high-fat feed induction.

Data Normality Test
To perform the Anova test, the requirements must be met, namely the data must be normally distributed After it was found that the data were normally distributed and had homogeneous variances, the Anova test could be performed.

One W ay Anova test
All the Anova test requirements have been fulfilled, namely consisting of normally distributed data and homogeneous data variance, then the One Way Anova test can be carried out.
From Table 3, the results of the One Way Anova test showed that spermatozoa motility had a significance value of less than 0.05 (p <0.05). It can be concluded that there were significant differences in at least two groups.

Post Hoc Test
To find out which groups had significant differences with other groups, a post hoc analysis was carried out.  Source: Primary Data, 2020

Discussion of Control Group Results
The results showed that a diet high in fat will cause damage to the spermatozoa motility of Wistar rats. This sperm morphology 25 .

Discussion of Treatment Group Results
Research in the treatment group given red dragon The results showed that the red dragon fruit extract at a dose of 60 mg / 200 grBB / day for 52 days and 104 days could be a source of antioxidants. This is because red dragon fruit has phenol and vitamin C antioxidants. Phenolic compounds can stabilize free radicals by quickly giving hydrogen atoms to free radicals, while radicals derived from antioxidant phenol compounds will be more stable than free radicals.
Phenolic compounds also have the ability to negate perioxide radicals and free radicals so they are effective in inhibiting lipid oxidation 27 . Vitamin C has been pr oven to be a strong antioxidant 28 . Vitamin C or Lascorbic acid contained in red dragon fruit is one of the antioxidants that acts as an ROS scavenger and has high polarity properties because it contains a lot of hydroxyl groups so that it is easily dissolved in water.
That vitamin C can re act and is also able to neutralize free radicals, both in terms of induced DNA damage and excessive ROS production, so that vitamin C can improve sperm quality 29 . Red dragon fruit has high levels of ascorbic acid, where ascorbic acid also has an important role for DNA protection in sperm 30 .