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Rotenone-induced necrosis in insect cells via the cytoplasmic membrane damage and mitochondrial dysfunction
Zhipeng Sun1, Li Xue2, Yun Li1
1Key Laboratory of Crop Integrated Pest Management in South China, Ministry of Agriculture and Rural Affairs, South China Agricultural University, Guangzhou 510642, China; Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou 510642, China.
Abstract:
Rotenone, a selective inhibitor of mitochondrial complex I, has been extensively studied on kinds of neuron and neuroblast in Parkinson's disease. However, little is known about the potential mechanism of this promising botanical insecticide upon insect cells. In the article, cell proliferation of two Lepidoptera cell lines, Spodoptera litura SL-1 cells and Spodoptera frugiperda Sf9 cells, were all inhibited by rotenone in a time- and dose-dependent manner. Typical necrotic characteristics of cell morphology and ultrastructure, such as plasma membrane collapses and organelle lyses, were all observed by transmission electron microscope and scanning electron microscope. Moreover, irregular DNA degradation was also detected by DNA gel electrophoresis and Hoechst 33258 staining, while the typical apoptotic feature, DNA ladder, hadn't been observed. Flow cytometric analysis showed that rotenone-induced cell death of Sf9 and SL-1 cells accompanied with the plasma membrane potential depolarization and mitochondrial membrane potential reduction. Furthermore, the activity of Na+-K+-ATPase was detected in our study. In conclusion, rotenone could cause necrosis but not apoptosis in insect cells through a mitochondrial- and plasmic membrane-dependent pattern, which shed a light on the rotenone-induced cytotoxicity on insects.
Insights
Rotenone, a mitochondrial complex I inhibitor, causes necrosis, not apoptosis, in insect cells. This botanical insecticide disrupts cell membranes and mitochondria, impacting insect health.
Area of Science:
- Insect toxicology
- Cell biology
- Biochemistry
Background:
- Rotenone is a known inhibitor of mitochondrial complex I, extensively studied in Parkinson's disease research.
- Its precise mechanism of action on insect cells remains largely uncharacterized.
Purpose of the Study:
- To investigate the cytotoxic effects and cell death mechanisms of rotenone in Lepidoptera insect cell lines.
- To elucidate the role of mitochondrial and plasma membranes in rotenone-induced toxicity.
Main Methods:
- Utilized Spodoptera litura (SL-1) and Spodoptera frugiperda (Sf9) cell lines.
- Assessed cell proliferation, morphology (light and electron microscopy), DNA degradation (gel electrophoresis, Hoechst staining), and membrane potential (flow cytometry).
- Measured Na+-K+-ATPase activity.
Main Results:
- Rotenone inhibited cell proliferation in a time- and dose-dependent manner.
- Observed necrotic cell morphology, irregular DNA degradation (no DNA ladder), plasma membrane depolarization, and reduced mitochondrial membrane potential.
- Detected alterations in Na+-K+-ATPase activity.
Conclusions:
- Rotenone induces necrosis, not apoptosis, in insect cells.
- The cell death pathway is dependent on mitochondrial and plasma membrane integrity.
- Findings provide insight into rotenone's insecticidal cytotoxicity.
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