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Immunomodulatory Effect of Parkia Speciosa Hassk

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Immunomodulatory Effect of Parkia Speciosa Hassk © 2020 Journal of Pharmacy & Pharmacognosy Research, 8 (5), 457-465, 2020 ISSN 0719-4250 http://jppres.com/jppres Original Article | Artículo Original Immunomodulatory effect of Parkia speciosa Hassk. pods extract on rat induced by Salmonella typhimurium [Efecto inmunomodulador del extracto de vainas de Parkia speciosa Hassk. en ratas inducidas por Salmonella typhimurium] Fitrya Fitrya1*, Annisa Amriani1, Rennie Puspa Novita1, Elfita2, Dewi Setiorini1 1Pharmacy Department, Sriwijaya University, Palembang, Indonesia. 2Chemistry Department, Sriwijaya University, Palembang, Indonesia. *E-mail: [email protected] Abstract Resumen Context: Parkia speciosa, which has phenolic compounds and Contexto: Parkia speciosa, que tiene compuestos fenólicos y actividades pharmacology activities, is a plant having potential immunomodulatory farmacológicas, es una planta que tiene agentes inmunomoduladores agents from the extract of its pod. potenciales del extracto de su vaina. Aims: To evaluate the potential of P. speciosa as an immunomodulatory Objetivos: Evaluar el potencial de P. speciosa como agente agent tested on albino rats induced by the bacteria Salmonella inmunomodulador en ratas albinas inducidas por la bacteria Salmonella typhimurium leading to typhoid fever. typhimurium que conduce a la fiebre tifoidea. Methods: The animals were divided into six groups: normal non-infected Métodos: Los animales se dividieron en seis grupos: grupo normal no group, negative control group, positive control, and three test groups infectado, grupo de control negativo, control positivo y tres grupos de with doses of 200, 400 and 800 mg/kg BW. The test groups were treated prueba con dosis de 200, 400 y 800 mg/kg de peso corporal. Los grupos by the extract for 12 days. The control and test groups were induced de prueba fueron tratados con el extracto durante 12 días. El control y with the S. typhimurium on the 8th day. On day 12th, CD4+, total los grupos de prueba fueron inducidos con S. typhimurium en el octavo leukocytes, differential cell count, and histology of the spleen organ día. El día 12, se evaluaron los CD4+, los leucocitos totales, el recuento were evaluated. Statistical analysis was performed by using one-way diferencial de células y la histología del órgano del bazo. El análisis ANOVA followed post hoc LSD test (α 0.05). estadístico se realizó mediante el uso de ANOVA unidireccional seguido Results: The result showed that the ethanol extract of P. speciosa pod de la prueba post hoc LSD (α 0,05). could increase the number of CD4+. The extract to 200 and 400 mg/kg Resultados: El extracto etanólico de la vaina de P. speciosa podría BW doses increased in the parameters of leukocyte, neutrophil, aumentar el número de CD4+. El extracto a dosis de 200 y 400 mg/kg de lymphocyte and monocyte with insignificant differences compared to peso corporal aumentó en los parámetros de leucocitos, neutrófilos, that of the positive control group (p>0.05). However, there were linfocitos y monocitos con diferencias insignificantes en comparación significant differences between the two first parameters between the test con el grupo de control positivo (p>0,05). Sin embargo, hubo diferencias group with 800 mg/kg BW doses compared to the positive control significativas entre los dos primeros parámetros entre el grupo de group. Moreover, more necroses in the spleen were observed from prueba con dosis de 800 mg kg de peso corporal en comparación con el histological analysis on 800 mg/kg BW doses than 400 mg/kg BW. grupo de control positivo. Además, se observaron más necrosis en el Conclusions: In general, P. speciosa ethanolic extract can increase the bazo a partir del análisis histológico con dosis de 800 mg/kg de peso immune system based on the CD4+ count, leukocyte, monocyte, and corporal que con 400 mg/kg de peso corporal. neutrophil, and causes normal necroses in low doses. Conclusiones: En general, el extracto etanólico de P. speciosa puede aumentar el sistema inmunitario en función del recuento de CD4+, leucocitos, monocitos y neutrófilos, y causa necrosis normales en dosis bajas. Keywords: Parkia speciosa; immunomodulator; Salmonella typhimurium; Palabras Clave: Parkia speciosa; inmunomodulador; Salmonella typhoid fever. typhimurium; fiebre tifoidea. ARTICLE INFO Received: March 21, 2020. Received in revised form: May 29, 2020. Accepted: May 31, 2020. Available Online: june 11, 2020. Declaration of interests: The authors declare no conflict of interest. Funding: This work was supported by funding from Research and Community Service Institution of Sriwijaya University (Indonesia) by mean the project “Hibah Penelitian Unggulan Kompetitif 2019” [No. 0149.159/UN9/SB3.LP2M.PT/2019]. _____________________________________ Fitrya et al. Immunomodulator effect of Parkia speciosa INTRODUCTION ed as an antioxidant, anti-carcinogenic and anti- inflammatory (Kamisah et al., 2013; Mustafa et al., Infectious diseases are the second leading cause 2018), antianemia (Nursucihta et al., 2014), antihy- of death in worldwide after cardiovascular disease pertensive and heart problems (Siow and Gan, (WHO, 2004). Infection can be caused by viruses, 2013; Kamisah et al., 2017) and antibacterial (Uyub bacteria, protozoa, worms, and parasitic fungi en- et al., 2010). However, the immunomodulatory tering the body or growing on the body surface. activity of P. speciosa has not been found in previ- An infectious disease with a high occurrence in ous studies. Thus, this study aims to reveal the several Asian countries is typhoid fever (Purba et potential of P. speciosa as an immunomodulatory al., 2016). This acute fever is caused by bacterium agent by observing CD4+ parameters, total leuko- Salmonella enterica infection, especially its deriva- cytes, differential blood cell counts, and spleen’s tive, Salmonella typhimurium (Alba et al., 2016). evaluation of albino rats induced by Salmonella Based on WHO data, the number of typhoid fever typhimurium bacteria. patients in Indonesia is relatively high, reaching 81,000 of 100,000 inhabitants (Rahmasari and Les- MATERIAL AND METHODS tari, 2018). A person with a weak immune system is at a high risk of developing typhoid fever (Kalia Vegetal material and chemicals et al., 2016). P. speciosa was collected from Musi Rawas dis- The innate immune system, which consists of tricts, South Sumatera, Indonesia (2.90°S, phagocyte cells, is the most important to defense 103.28°E). This species has been identified by the against microorganisms and malignant cells (Ha- Indonesian Institute of Science with Register No. run et al., 2015; Venkatalakshmi et al., 2016). Acti- 218/IPH.06/HM/I/2018. Salmonella typhimurium vation of the immune system is required to help a ATCC 14028 (105 CFU) was obtained from the body destroying antigenic substances, and it can Central Laboratory of Health, Palembang, South be stimulated using immunomodulators. Sumatera, Indonesia. Chemicals as methanol, Synthetic immunomodulatory drugs have NaCl (Merck), phosphate buffer pH 6,8 and Giem- many benefits, but adverse side effects limit their sa stain (Sigma-Aldrich) were in an analytical use. Further studies are thus an important way to grade. find safer and more effective immunomodulatory Preparation of P. speciosa pod extract agents (Venkatalakshmi et al., 2016). Plant extracts are commonly considered as potential agents from Ethanol extract of P. speciosa pod was prepared their immunomodulatory properties, which have by a maceration method. Its powder (500 mg) was smaller side effects (Alamgir and Uddin, 2010). soaked with 2 L of ethanol 95% for 2 days. A re- Their properties are generated by activating a maceration process of the residue was carried out function and efficiency of macrophages, granulo- twice. The maceration result then was evaporated cytes, complement, and natural killer cells and by with a rotary evaporator (Yamato®) at a tempera- producing effector molecules (jayathirtha and ture of 65°C to get a concentrated extract (Harun et Mishra, 2004). Plant metabolites such as sterols, al., 2015). polysaccharides, alkaloids, flavonoids, lectins, and glycoproteins are used as immunomodulatory Design of test animal experiments agents (Harun et al., 2015). The pod of Parkia speci- Wistar male white albino rats (150 - 200 g osa Hassk. (Leguminosae) contains phytochemical weight) were used in this study. An ethics certifi- compounds such as flavonoids, phenolics, alka- cate for the use of test animals was obtained from loids, and saponins (Kamisah et al., 2013). Phar- the Faculty of Medicine, Sriwijaya University, with macology activities of P. speciosa have been report- No. 064/kepkrsmhfkunsri/2019. The test animals http://jppres.com/jppres J Pharm Pharmacogn Res (2020) 8(5): 458 Fitrya et al. Immunomodulator effect of Parkia speciosa were acclimatized to the laboratory environment cells were manually analyzed by peripheral blood at a temperature of 23 ± 2°C and a 12-hour light- smear observation under a microscope (Olympus dark cycle for one week. During the acclimation CX21®). process, these rats were given standard ad libitium drink and feed (johnson et al., 2017). They were Evaluation of spleen organs divided into 6 groups: the normal group and nega- Peritoneal and thoracic cavities were opened af- tive control group were given Tween-80 0.5 ter euthanizing by cervical dislocation. Spleen or- mL/kg BW; the positive control group was given gans were isolated and washed with distilled water 0.54 mL (dose 2.5 mg/kg BW) of Stimuno® syrup and 0.9% NaCl, consecutively. A macroscopic ex- (containing extracts of Centella asiatica, 5 mg/mL); amination was observed based on size, weight, the test groups I-III received 200, 400 and 800 color, and consistency of spleen organs. Then the mg/kg BW of the extract, respectively. All groups spleen was fixed with 10% formalin for histologi- (except the normal group) were also given 0.5 mL cal analysis. Salmonella typhimurium 105 CFU (intraperitoneal) on the 8th day of treatment as the agent inducer of Statistical analysis the typhoid fever. The CD4+ and the differential counts were ana- Antigen preparation lyzed using SPSS for Windows. Macroscopic find- ings of the spleen of each test group were com- S.
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