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Background: The regulation of microglial function via mitochondrial homeostasis is important in the development of neuroinflammation. The underlying mechanism for this regulatory function remains unclear. In this study, we investigated the protective role of mitochonic acid 5 (MA-5) in microglial mitochondrial apoptosis following TNFα-induced inflammatory injury. Methods: TNFα was used to induce inflammatory injury in mouse microglial BV-2 cells with and without prior MA-5 treatment. Cellular apoptosis was assessed using the MTT and TUNEL assays. Mitochondrial functions were evaluated via mitochondrial membrane potential JC-1 staining, ROS flow cytometry analysis, mPTP opening assessment, and immunofluorescence of cyt-c. Mitophagy was examined using western blots and immunofluorescence. The pathways analysis was carried out using western blots and immunofluorescence with a pathway blocker. Results: Our results demonstrated that TNFα induced apoptosis in the microglial BV-2 cell line by activating the caspase-9-dependent mitochondrial apoptotic pathway. Mechanistically, inflammation reduced mitochondrial potential, induced ROS production, and contributed to the leakage of mitochondrial pro-apoptotic factors into the cytoplasm. The inflammatory response reduced cellular energy metabolism and increased oxidative stress. By contrast, treatment with MA-5 reduced mitochondrial apoptosis via upregulation of mitophagy. Increased mitophagy degraded damaged mitochondria, disrupting mitochondrial apoptosis, neutralizing ROS overproduction, and improving cellular energy production. We also identified that MA-5 regulated mitophagy via Bnip3 through the MAPK–ERK–Yap signaling pathway. Inhibiting this signaling pathway or knocking down Bnip3 expression prevented MA-5 from having beneficial effects on mitochondrial homeostasis and increased microglial apoptosis. Conclusions: After TNFα-induced inflammatory injury, MA-5 affects microglial mitochondrial homeostasis in a manner mediated via the amplification of protective, Bnip3-related mitophagy, which is mediated via the MAPK–ERK–Yap signaling pathway.
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Ethnobotany of Acorus in China

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In China, species in the genus Acorus are used for many different purposes, viz., in traditional medicine, for rituals, and as ornamental plants. Acorus calamus (sweet flag) has been a symbolic plant in Chinese culture for many centuries and is used as a ceremonial object in festivals throughout the country. For other Acorus species in China, ethnobotanical records are few. During field investigations from 2006 to 2017, we interviewed 573 individuals representing ethnic groups such as the Miao, Yao, Buyi, Shui, Tujia, Dong, She, Maonan, Zhuang, Yi, and Han people. Various ethnobotanical approaches were adopted in the field surveys, including key informant interviews, semistructured interviews, participatory observation, direct observation, etc. The results have revealed a rich treasure trove of traditional knowledge on Acorus in China. Two species (A. macrospadiceus and A. gramineus) were used by ethnic groups as spices to add flavor to meat. Through the study found that all the species of Acorus had medicinal value, especially A. tatarinowii, deafness, blindness, and digestive disorders were among the conditions commonly treated using these plants. Traditional knowledge of the cultural, ethnomedicinal, and food values of Acorus, based on our recent literature surveys and field investigations in China is presented here. A better understanding of Acorus is vital for conserving the plants and the traditional knowledge associated with them.
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