Original Article Jatonik polyherbal mixture induced rat liver MMPT pore opening in normal Wistar rat: In vitro and in vivo studies Olabinri P. Folashade a , Ibrahim Damilare Boyenle a,c , Tolulope A. Oyedeji d , Fiyinfoluwa Demilade Ojeniyi b , Adisa Ayobami Damilare a , Leonard O. Ehigie b , Adeola Folasade Ehigie a,⇑ a Membrane Biochemistry and Biophysics Research Laboratory, Department of Biochemistry, Ladoke Akintola University of Technology, Ogbomoso 210214, Nigeria b Computational Biology/Drug Discovery Laboratory, Department of Biochemistry, Ladoke Akintola University of Technology, Ogbomoso 210214, Nigeria c College of Health Sciences, Crescent University Abeokuta, Abeokuta 111105, Nigeria d Department of Biochemistry, University of Lagos, Lagos 101017, Nigeria article info Article history: Received 2 December 2022 Revised 6 April 2023 Accepted 19 June 2023 Available online xxxx Keywords: Acute toxicity Ageratum conyzoides Linn. Apoptosis Hunteria umbellate (K. Schum.) Hallier f. Lepidium meyenii Walp. Mitochondrial ATPase Mitochondrial lipid peroxidation Mitochondrial membrane permeability transition pore Polyherbal mixture Xylopia aethiopica (Dunal) A. Rich abstract Objective: To assess acute toxicity, the in vitro and in vivo effects of methanol and ethyl acetate extracts (JME and JEE) of Jatonik polyherbal mixture on some mitochondria-related parameters and their effect on the activity of some liver enzymes. Methods: Acute toxicity of JME and JEE was determined using Lorke’s method. In vitro and in vivo opening of the mitochondrial membrane permeability transition pore (MMPT pore) was spectrophotometrically assayed. Production of malondialdehyde (MDA) as an index of lipid peroxidation and the activity of mito- chondrial ATPase was evaluated in vitro and in vivo and the effect of JME and JEE on the activity of liver enzymes: alkaline phosphatase (ALP), aspartate and alanine aminotransferase (AST and ALT) and gamma- glutamyl transferase (GGT) activities were also investigated. Results: JME had an LD 50 of 3808 mg/kg b.w whereas JEE had an LD 50 greater than 5000 mg/kg b.w. of rats. After the rats have been fed with both extracts, a photomicrograph of a piece of liver tissue showed no apparent symptoms of toxicity. From the in vitro and in vivo studies, both extracts prompted intact mitochondria to open their MMPT pores. When compared to the control, lipid peroxide product release and ATPase activity were significantly increased (P < 0.05) in vitro and in vivo. The activities of AST, ALT, and GGT were all reduced at 50 mg/kg when treated with JME, but the activity of AST was considerably enhanced when treated with JEE (P < 0.05). The results revealed that both JME and JEE of the Jatonik poly- herbal mixture had low toxicity, profound MMPTpore induction, and enhanced ATPase activity, but an increased MDA production. Conclusion: Jatonik extracts may be a promising target for drug development in diseases where there is dysregulation of apoptosis, however, further studies are needed to better clarify the molecular mecha- nism involved in these phenomena. Ó 2023 Tianjin Press of Chinese Herbal Medicines. Published by ELSEVIER B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). 1. Introduction Cell death is an important biological process that can be induced by intrinsic or extrinsic apoptosis, and often time abnor- malities in its pathway can contribute to pathologies in human dis- ease (Lin et al., 2014). Mitochondria is a dynamic organelle that produces ATP which is also an important contributor to cell activ- ities in both development and cell death processes. Mitochondrial permeability transition pore (mPTP) regulates this dual function of mitochondria (Pérez & Quintanilla, 2017) and hence its involve- ment in apoptotic cell death via the formation of a nonspecific pore formed from a voltage–dependent anionic channel (VDAC), the adenine nucleotide translocase and cyclophilin-D (cyP-D) at con- tact sites between mitochondrial outer and inner membranes (Crompton, 1999). Mitochondria permeability transition (mPT) which is mediated by the mitochondrial permeability transition pore (mPTP) causes the permeabilization of the outer membrane of the mitochondria (Bonora et al., 2021) and the opening of a non- specific pore in the inner membrane of the mitochondria, convert- ing them from organelles whose production of ATP sustains the cell to instruments of death (Halestrap, 1999). Opening of the mPTP can be triggered by calcium overload that was stimulated by mito- chondria metabolism (Zhou et al., 2022) and this allows the release https://doi.org/10.1016/j.chmed.2023.06.002 1674-6384/Ó 2023 Tianjin Press of Chinese Herbal Medicines. Published by ELSEVIER B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). ⇑ Corresponding author. E-mail address: afehigie@lautech.edu.ng (A.F. Ehigie). Chinese Herbal Medicines xxx (xxxx) xxx Contents lists available at ScienceDirect Chinese Herbal Medicines journal homepage: www.elsevier.com/locate/chmed Please cite this article as: O.P. Folashade, I.D. Boyenle, T.A. Oyedeji et al., Jatonik polyherbal mixture induced rat liver MMPT pore opening in normal Wistar rat: In vitro and in vivo studies, Chinese Herbal Medicines, https://doi.org/10.1016/j.chmed.2023.06.002