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      Effect of Mahuang Gancao Ganjiang Decoction on Fusion and Fission of Mitochondria and Apoptosis of Lymphocytes in Mice under Cold Stress

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          Abstract

          Mahuang Gancao Ganjiang Decoction (MGGD) can effectively alleviate the symptoms of the patients suffering from exogenous cold stress. However, the curative mechanism has not been fully clarified. This study was designed to investigate the effect of MGGD on the apoptosis of lymphocytes induced by cold stress in mice. The model mice were randomly divided into four groups: the normal control group (no handling mice), cold stress group, MGGD + cold stress group, and MGGD group. Lymphocytes of the mice were isolated from the peripheral blood. Electron microscopy analysis revealed cold stress resulted in mitochondrial fragmentation. Accompanied with the change of morphology of mitochondria, ATP production and the activity of respiratory chain complex decreased in these cells. Western blot analysis showed that these cells expressed decreased fusion-related proteins Mitofusin 1 (Mfn1), Mitofusin 2 (Mfn2), and optic atrophy protein-1 (Opa-1) and increased fission-related proteins dynamin-related protein 1 (Drp1) and fission 1 (Fis-1); our results also show that decreased mitochondrial fusion induces cell apoptosis during cold stress. Meanwhile, we found MGGD can inhibit cell apoptosis induced by cold stress through regulating expression level of Mfn1, Mfn2, Drp1, Fis-1, and Opa-1. These findings are very significant for understanding how MGGD regulates cold-stress-induced cell apoptosis.

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          Most cited references20

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          C. elegans Dynamin-Related Protein DRP-1 Controls Severing of the Mitochondrial Outer Membrane

          Little is known about the mechanism of mitochondrial division. We show here that mitochondria are disrupted by mutations in a C. elegans dynamin-related protein (DRP-1). Mutant DRP-1 causes the mitochondrial matrix to retract into large blebs that are both surrounded and connected by tubules of outer membrane. This indicates that scission of the mitochondrial outer membrane is inhibited, while scission of the inner membrane still occurs. Overexpressed wild-type DRP-1 causes mitochondria to become excessively fragmented, consistent with an active role in mitochondrial scission. DRP-1 fused to GFP is observed in spots on mitochondria where scission eventually occurs. These data indicate that wild-type DRP-1 contributes to the final stages of mitochondrial division by controlling scission of the mitochondrial outer membrane.
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            Mitochondria are morphologically and functionally heterogeneous within cells.

            We investigated whether mitochondria represent morphologically continuous and functionally homogenous entities within single intact cells. Physical continuity of mitochondria was determined by three-dimensional reconstruction of fluorescence from mitochondrially targeted DsRed1 or calcein. The mitochondria of HeLa, PAEC, COS-7, HUVEC, hepatocytes, cortical astrocytes and neuronal cells all displayed heterogeneous distributions and were of varying sizes. There was a denser aggregation of mitochondria in perinuclear positions than in the cell periphery, where individual isolated mitochondria could be seen clearly. Using fluorescence-recovery after photobleaching, we observed that DsRed1 and calcein were highly mobile within the matrix of individual mitochondria, and that mitochondria within a cell were not lumenally continuous. Mitochondria were not electrically coupled, since only individual mitochondria were observed to depolarize following irradiation of TMRE-loaded cells. Functional heterogeneity of mitochondria in single cells was observed with respect to membrane potential, sequestration of hormonally evoked cytosolic calcium signals and timing of permeability transition pore opening in response to tert-butyl hydroperoxide. Our data indicate that mitochondria within individual cells are morphologically heterogeneous and unconnected, allowing them to have distinct functional properties.
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              A common lipid links Mfn-mediated mitochondrial fusion and SNARE-regulated exocytosis.

              Fusion of vesicles into target membranes during many types of regulated exocytosis requires both SNARE-complex proteins and fusogenic lipids, such as phosphatidic acid. Mitochondrial fusion is less well understood but distinct, as it is mediated instead by the protein Mitofusin (Mfn). Here, we identify an ancestral member of the phospholipase D (PLD) superfamily of lipid-modifying enzymes that is required for mitochondrial fusion. Mitochondrial PLD (MitoPLD) targets to the external face of mitochondria and promotes trans-mitochondrial membrane adherence in a Mfn-dependent manner by hydrolysing cardiolipin to generate phosphatidic acid. These findings reveal that although mitochondrial fusion and regulated exocytic fusion are mediated by distinct sets of protein machinery, the underlying processes are unexpectedly linked by the generation of a common fusogenic lipid. Moreover, our findings suggest a novel basis for the mitochondrial fragmentation observed during apoptosis.
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                Author and article information

                Journal
                Evid Based Complement Alternat Med
                Evid Based Complement Alternat Med
                ECAM
                Evidence-based Complementary and Alternative Medicine : eCAM
                Hindawi Publishing Corporation
                1741-427X
                1741-4288
                2017
                16 January 2017
                : 2017
                : 5132963
                Affiliations
                Hubei University of Chinese Medicine, Basic Medical College, Wuhan 430065, China
                Author notes

                Academic Editor: Luciana Dini

                Author information
                http://orcid.org/0000-0002-4905-3096
                http://orcid.org/0000-0002-2079-873X
                Article
                10.1155/2017/5132963
                5278204
                69b3d84a-0d1f-466d-9935-9ba6e82e4f8b
                Copyright © 2017 L. Chen and H. Chen.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 4 September 2016
                : 23 November 2016
                : 21 December 2016
                Funding
                Funded by: Hubei Province Education Department of China
                Award ID: D20152002
                Award ID: B2016112
                Categories
                Research Article

                Complementary & Alternative medicine
                Complementary & Alternative medicine

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