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1: # Effects of α-mangostin against rotenone-induced Parkinson’s disease-like symptoms in mice 2: Author: Yung-Jung Tsai, Yu-Hsin Lai 3: Date: 10-6-2021 4: Link: https://doi.org/10.1186/s12906-021-03466-z 5: BMC Complementary Medicine and Therapies: Research 6: ## Abstract 7: BackgroundRotenone-induced neurotoxicity has been used as an animal model for Parkinson’s disease (PD). α-Mangostin has been reported to exhibit protective effects against rotenone-induced toxicity. 8: MethodsIn this study we investigated whether α-mangostin could protect mice against rotenone-induced PD-like symptoms. 9: ResultsRotenone exposure increased dopaminergic neuron loss in substantia nigra pars compacta (SNpc), tyrosine hydroxylase immunoreactivity (TH-IR) density decrease in striatum (STR), decreased striatal dopamine content and increased microglial activation marker ionized calcium-binding adaptor molecule-1 (Iba-1) immunoreactivity density compared with vehicle-treated controls. 10: Oral administration of α-mangostin at doses of either100 mg/kg/day or200 mg/kg/day reversed these effects. 11: In addition α-mangostin treatment decreased oxidative stress marker nitrotyrosine immunoreactivity density but had no effect on protein levels of anti-apoptotic gene B-cell lymphoma-extra large(Bcl-xl)and pro-apoptotic gene Baxin SNpc. 12: ConclusionThese results indicate that α-mangostin can protect against rotenone-induced PD-like symptoms via reducing oxidative stress. 13: ## Background 14: Parkinson’s disease (PD) is one of the most common neurodegenerative disorders worldwide [1]. The most characteristic pathological symptom is motor dysfunction resulting from degeneration of dopaminergic neurons in substantia nigra pars compacta (SNpc) [2]. The pathogenesis remains unclear; however oxidative stress [3]and mitochondrial dysfunction [4]have been proposed as possible causes. 15: Rotenone has been used as an insecticide since ancient times [5]. In humans chronic exposure has been linked with development of PD [6]. Mice exposed chronically have shown similar symptoms [7],and thus rotenone-induced PD mouse models have been widely used in studies investigating PD pathogenesis [8]. 16: The mangosteen fruit belongs to Garcinia species [9]. Its extracts have exhibited antioxidant [10], anti-inflammatory [11]and neuroprotective effects [12]. One major active compound found within mangosteen fruit extracts is α-mangostin [13],which has been reported to exhibit protective effects against rotenone-induced toxicity [14]. 17: In this study we examined whether oral administration of α-mangostin could protect against rotenone-induced PD-like symptoms. 18: ## Methods 19: ### Animals 20: All animal experiments were conducted according to National Institutes of Health Guide for Care and Use of Laboratory Animals under approval from Institutional Animal Care Committee at Kaohsiung Medical University(KMU). 21: Eight-week-old male C57BL/6 mice were purchased from BioLASCO Taiwan Co., Ltd., Taiwan(Stock No.: BL-006)and maintained under standard conditions(temperature at25 ± 1 °C,humidity at60 ± 5%and12-h light-dark cycle). Food was provided ad libitum except during experimental procedures. 22: ### Experimental design 23: Thirty-two mice were randomly assigned into four groups(each containing eight mice): Control group(mice treated intraperitoneally(IP)with saline solution(0.9% NaCl)daily); Rotenone group(mice treated IP with rotenone(1 mg/kg/day)daily); Low-dose α-mangostin group(mice treated orally(LPO)with α-mangostin(100 mg/kg/day)daily); High-dose α-mangostin group(mice treated LPOwith α-mangostin(200 mg/kg/day)daily). 24: All treatments lasted for five weeks. 25: Mice were sacrificed after overnight fasting under ketamine/xylazine anesthesia at end point. 26: ### Rotenone preparation 27: Rotenone was purchased from Sigma-Aldrich Chemical Co.(St.Louis,Missouri,U.S.A.)and dissolved at concentrationof10 mg/ml in DMSO(Sigma-Aldrich Chemical Co.,St.Louis,Missouri,U.S.A.)followed by dilution at ratioof1:9in saline solution(0.9% NaCl). 28: ### Preparationofα-mangostin 29: α-Mangostin was purchased from Sigma-Aldrich Chemical Co.(St.Louis,Missouri,U.S.A.). The powder was dissolved at concentrationof50 mg/ml in DMSO(Sigma-Aldrich Chemical Co.,St.Louis,Missouri,U.S.A.)followed by dilution at ratioof1:9with Tween80(Sigma-Aldrich Chemical Co.,St.Louis,Missouri,U.S.A.)for oral administration. 30: ### Tissue preparation 31: Mice were sacrificed under ketamine/xylazine anesthesia after overnight fasting at end point. 32: Brains were rapidly removed then immersed immediately into ice-cold fixative solution containing phosphate-buffered saline(PBS)(Gibco Thermo Fisher Scientific,Waltham,Massachusetts,U.S.A.)and paraformaldehyde(4%)for postfixation overnight followed by immersion into PBS containing sucrose(30%)for cryoprotection. 33: ### Immunohistochemistry 34: Cryoprotected brains were cut into coronal sections(30 μm thickness)using cryostat microtome(LS1510,Thermo Fisher Scientific,Waltham,Massachusetts,U.S.A.). 35: Sections were incubated overnightat4 °Cwith primary antibodies rabbit anti-ionized calcium-binding adaptor molecule-1(Iba-1)(1/1000;Wako Pure Chemical Industries,Ltd.,Osaka,Japan), rabbit anti-nitrotyrosine(NT)(1/500;Abcam plc,Cambridge,Cambridgeshire,U.K.), rabbit anti-Bcl-xl(1/500;Abcam plc,Cambridge