Chlorpyrifos induces apoptosis and autophagy in common carp lymphocytes by influencing the TCR γ-dependent PI3K/AKT/JNK pathway

Abstract

Chlorpyrifos is an insecticide that is widely used in agricultural production. However, little is known about how chlorpyrifos disrupts lymphocyte homeostasis in common carp. Herein, we identified TCRγ through the results of transcriptome analysis. Subsequently, we established TCR γ knockdown and overexpression models in carp head kidney lymphocyte respectively using RNA interference and the pcDNA3.1 plasmid, respectively. Real-time PCR, fluorescent staining, ultrastructure observation and flow cytometry were used to detect the levels of the PI3K/AKT pathway, autophagy and apoptosis. Our results demonstrated that chlorpyrifos significantly decreased the expression of TCR γ, TCR γ suppression thereby induced increased mRNA expression of TNF-α, Bax, caspase-3, caspase-8, caspase-9 and significantly inhibited the expression of Bcl-2, which indicated that apoptosis was triggered. This conclusion was supported by our flow cytometry and ultrastructure observation results. In addition, the control and TCR γ overexpression groups had normal cell morphology. Moreover, TCR γ suppression activated the expression of Becline-1, ATG5, ATG10, ATG12, ATG16 and reduced the expression of mTOR, with the opposite results observed in the TCR γ overexpression group. Together, these results suggested that TCR γ imbalance triggers apoptosis and autophagy in lymphocyte. Moreover, we found that TCR γ knockdown significantly increased the mRNA expression of JNK and decreased the expression of PI3K and AKT, which indicated that the PI3K/AKT/JNK pathway was activated. Our results reported here indicated that chlorpyrifos induces apoptosis and autophagy in head kidney lymphocyte through the inhibition of TCR γ.