The Effect of Javanese Chili (Piper retrofractum) and Lemon (Citrus limon) Formula on Total Cholesterol Levels in Male White Rats
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CHOD-PAP, Total Cholesterol, Piper retrofractum, Citrus limon, cholesterol-cholate-thiouracil
Abstract
Hypercholesterolemia, defined as blood cholesterol levels exceeding 200 mg/dL, is a significant risk factor for cardiovascular disease. Traditional Indonesian medicinal plants, such as Javanese chili (Piper retrofractum) and lemon (Citrus limon), have long been recognized for their potential lipid-lowering properties. This study aimed to scientifically evaluate the hypocholesterolemic effect of a combined P. retrofractum and C. limon extract in a hypercholesterolemic rat model. Thirty male Sprague-Dawley rats were randomly assigned to seven groups: a normal control, a negative control (Na-CMC), a positive control (simvastatin 0.018 mg/200 g BW), a scientific herbal formula (0.18 g/200 g BW), and three treatment groups receiving P. retrofractum and C. limon in ratios of 1:1, 1:2, and 2:1. Hypercholesterolemia was induced using a cholesterol-cholate-thiouracil (CCT) mixture alongside a high-fat diet. Total cholesterol levels were quantified using the Cholesterol Oxidase-Peroxidase Aminoantipyrine (CHOD-PAP) method. After 21 days of treatment, all P. retrofractum and C. limon-treated groups demonstrated a statistically significant reduction in total cholesterol compared to the negative control (p <0.05). Notably, the 1:1 and 1:2 ratios of P. retrofractum to C. limon yielded the most substantial cholesterol reductions (44% and 43%, respectively), comparable to the scientific herbal formula and superior to simvastatin (30%). In contrast, the 2:1 ratio showed a comparatively lower reduction (38%), suggesting a potential inhibitory effect of excessive P. retrofractum on C. limon's lipid-lowering properties. These findings collectively highlight the promising potential of optimized P. retrofractum and C. limon combinations, particularly at 1:1 and 1:2 ratios, as effective natural interventions for managing hypercholesterolemia.
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References
2. Aisyah YL, Ilmi IMB, Marjan AQ. Factors Affecting Total Cholesterol Level in the Pre-Elderly and Elderly in Kedaung Village. J Kesmas Jambi. 2025;9(1):1-8. DOI: 10.22437/jkmj.v9i1.41685
3. Khatiwada N, Hong Z. Potential Benefits and Risks Associated with the Use of Statins. Pharmaceutics. 2024;16(2):214. DOI: 10.3390/pharmaceutics16020214; PMCID: PMC10892755; PMID: 38399268
4. Azzahra F, Suleman HF, Yunas RW, Iswandana R. Wisdom of nature: Cardiovascular, anti-obesity, and immunomodulator nutraceuticals. J Appl Pharm Sci. 2025;15(1):43-63. DOI: 10.7324/JAPS.2024.196392
5. Faramayuda F, Permana J, Syam AK, Elfahmi E. Identification Secondary Metabolites from Callus Piper retrofractum Vahl. Elkawnie J Islamic Sci Technol. 2021;7(1):197-214. DOI: 10.22373/ekw.v7i1.8630
6. Klimek-Szczykutowicz M, Szopa A, Ekiert H. Citrus limon (Lemon) Phenomenon-A Review of the Chemistry, Pharmacological Properties, Applications in the Modern Pharmaceutical, Food, and Cosmetics Industries, and Biotechnological Studies. Plants. 2020;9(1):119. DOI: 10.3390/plants9010119; PMCID: PMC7020168; PMID: 31963590
7. Pratiwi A, Mustofa S, Susantiningsih T. Pengaruh Pemberian Ekstrak Etanol 95% Cabe Jawa (Piper reftrofractum Vahl.) terhadap Kadar HDL Tikus Putih (Rattus novergicus) Jantan Galur Sprague Dawley yang Diberi Diet Tinggi Lemak. Majority Med J Lampung Univ. 2014;3(1):1–8.
8. Rana R, Mehmood MH, Shaukat B, Shahid S, Malik A, Murtaza B. Evaluation of the Cardiovascular Effects of Coriandrum sativum and Citrus limon to Treat Arsenic-Induced Endothelial Damage and Hypertension in Rats. Life. 2022;12(11):1842. DOI: 10.3390/life12111842; PMCID: PMC9695286; PMID: 36362997
9. Hose. The Effect of Solvent Variations on the Phytochemical Profile of Java Chime Fruit Extract (Piper Retrofractum Vahl) With Ultrasonic Method. Interdisiplin J Qual Quant Res. 2025;2(2):79-97. DOI: 10.61166/interdisiplin.v2i2.60
10. Mayasari U, Laoli MT. Karakterisasi Simplisia dan Skrining Fitokimia Daun Jeruk Lemon (Citrus limon (L.) Burm.f.). Klorofil J Ilmu Biol Terapan. 2018;2(1):7-13. DOI: 10.30821/kfl:jibt.v2i1.1802
11. Panche AN, Diwan AD, Chandra SR. Flavonoids: an overview. J Nutr Sci. 2016;5:e47. DOI: 10.1017/jns.2016.41; PMCID: PMC5465813; PMID: 28620474
12. Safdar A, Iftikhar S, Karim M, Khan R, Hayat MQ, Afsar T, et al. Rapid and cost-effective colorimetric detection of alkaloids in botanical extracts using a novel protocol with compressed tablets. Sci Rep. 2025;15(1):26332. DOI: 10.1038/s41598-025-11907-w; PMCID: PMC12277426; PMID: 40685408
13. Hayat J, Akodad M, Moumen A, Baghour M, Skalli A, Ezrari S, et al. Phytochemical screening, polyphenols, flavonoids and tannin content, antioxidant activities and FTIR characterization of Marrubium vulgare L. from 2 different localities of Northeast of Morocco. Heliyon. 2020;6(11):e05609. DOI: 10.1016/j.heliyon.2020.e05609; PMCID: PMC7708819; PMID: 33305038
14. Adu JK, Amengor CDK, Kabiri N, Orman E, Patamia SAG, Okrah BK. Validation of a Simple and Robust Liebermann-Burchard Colorimetric Method for the Assay of Cholesterol in Selected Milk Products in Ghana. Int J Food Sci. 2019;2019:9045938. DOI: 10.1155/2019/9045938; PMCID: PMC6815567; PMID: 31737650
15. Dubale S, Kebebe D, Zeynudin A, Abdissa N, Suleman S. Phytochemical Screening and Antimicrobial Activity Evaluation of Selected Medicinal Plants in Ethiopia. J Exp Pharmacol. 2023;15:51-62. DOI: 10.2147/jep.s379805; PMCID: PMC9922502; PMID: 36789235
16. Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional. Tujuh Ramuan Jamu Saintifik: Pemanfaatan Mandiri oleh Masyarakat. Tawangmangu: Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional; 2017.
17. Fauzi NI, Ulfah M, Yunis YF. Efek Antiobesitas Ekstrak Etanol Bawang Dayak (Eleutherine bulbosa (Mill.) Urb) pada Tikus Model Obesitas. J Ilmiah Farmako Bahari. 2019;10(2):123–31. DOI: 10.52434/jfb.v10i2.653
18. Kurniati NF, Nurfatwa M, Artarini AA. Aktivitas Makrofag Pada Aorta Tikus Hiperkolesterolemia. Majalah Kedokteran Bandung. 2018;50(1):13–20. DOI: 10.15395/mkb.v50n1.1159
19. Widada ST, Martsiningsih MA, Carolina SC. Gambaran Perbedaan Kadar Kolesterol Total Metode CHOD-PAP (Cholesterol Oxidase - Peroxsidase Aminoantypirin) Sampel Serum dan Sampel Plasma EDTA. J Teknol Lab. 2016;5(1):41–4.
20. Nikolić MM, Jovanović KK, Marković TL, Marković DL, Gligorijević NN, Radulović SS, et al. Antimicrobial synergism and cytotoxic properties of Citrus limon L., Piper nigrum L. and Melaleuca alternifolia (Maiden and Betche) Cheel essential oils. J Pharm Pharmacol. 2017;69(11):1606-14. DOI: 10.1111/jphp.12792; PMID: 28815601
21. Zhang J, Lv C, Wang HN, Cao Y. Synergistic interaction between total glucosides and total flavonoids on chronic constriction injury induced neuropathic pain in rats. Pharm Biol. 2013;51(4):455–62. DOI: 10.3109/13880209.2012.739177; PMID: 23336442
22. Zulkarnain Z, Triyono A, Ardiyanto D, Saryanto S. Uji Klinik Keamanan Ramuan Jamu Penurun Kolesterol. J Kefarmasian Indones. 2021;11(1):8–16. DOI: 10.22435/jki.v11i1.3186
23. Ulfatun N, Saryanto, Wahyono S, Wijayanti E. Ramuan Jamu Saintifik Hiperkolesterolemia. In: Pramono S, Wijayahadi N, Rahardjo S, Sukasediati N, editors. Jamu Saintifik: Suatu Lompatan Ilmiah Pengembangan Jamu. Tawangmangu: Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional; 2017.
24. Cheung B, Sikand G, Dineen EH, Malik S, El-Farra AB. Lipid-Lowering Nutraceuticals for an Integrative Approach to Dyslipidemia. J Clin Med. 2023;12(10):3414. DOI: 10.3390/jcm12103414; PMCID: PMC10219430; PMID: 37240523
25. Rousset X, Vaisman B, Sethi AA, Remaley AT. Lecithin: cholesterol acyltransferase--from biochemistry to role in cardiovascular disease. Curr Opin Endocrinol Diabetes Obes. 2009;16(2):163-71. DOI: 10.1097/med.0b013e328329233b; PMCID: PMC2910390; PMID: 19306528
26. Elmaghraby DA, Alsalman GA, Alawadh LH, Al-Abdulqader SA, Alaithan MM, Alnuwaysir BI. Integrated traditional herbal medicine in the treatment of gastrointestinal disorder: the pattern of use and the knowledge of safety among the Eastern Region Saudi population. BMC Complement Med Ther. 2023;23(1):373. DOI: 10.1186/s12906-023-04197-0; PMCID: PMC10591345; PMID: 37872543
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