MD2 contributes to the pathogenesis of perioperative neurocognitive disorder via the regulation of α5GABA_A receptors in aged mice

Background: Perioperative neurocognitive disorder (PND) is a long-term postoperative complication in elderly surgical patients. The underlying mechanism of PND is unclear, and no effective therapies are currently available. It is believed that neuroinflammation plays an important role in triggering PND. The secreted glycoprotein myeloid differentiation factor 2 (MD2) functions as an activator of the Toll-like receptor 4 (TLR4) inflammatory pathway, and α5GABA_A receptors (α5GABA_ARs) are known to play a key role in regulating inflammation-induced cognitive deficits. Thus, in this study, we aimed to investigate the role of MD2 in PND and determine whether α5GABA_ARs are involved in the function of MD2.

Methods: Eighteen-month-old C57BL/6J mice were subjected to laparotomy under isoflurane anesthesia to induce PND. The Barnes maze was used to assess spatial reference learning and memory, and the expression of hippocampal MD2 was assayed by western blotting. MD2 expression was downregulated by bilateral injection of AAV-shMD2 into the hippocampus or tail vein injection of the synthetic MD2 degrading peptide Tat-CIRP-CMA (TCM) to evaluate the effect of MD2. Primary cultured neurons from brain tissue block containing cortices and hippocampus were treated with Tat-CIRP-CMA to investigate whether downregulating MD2 expression affected the expression of α5GABA_ARs. Electrophysiology was employed to measure tonic currents. For α5GABA_ARs intervention experiments, L-655,708 and L-838,417 were used to inhibit or activate α5GABA_ARs, respectively.

Results: Surgery under inhaled isoflurane anesthesia induced cognitive impairments and elevated the expression of MD2 in the hippocampus. Downregulation of MD2 expression by AAV-shMD2 or Tat-CIRP-CMA improved the spatial reference learning and memory in animals subjected to anesthesia and surgery. Furthermore, Tat-CIRP-CMA treatment decreased the expression of membrane α5GABA_ARs and tonic currents in CA1 pyramidal neurons in the hippocampus. Inhibition of α5GABA_ARs by L-655,708 alleviated cognitive impairments after anesthesia and surgery. More importantly, activation of α5GABA_ARs by L-838,417 abrogated the protective effects of Tat-CIRP-CMA against anesthesia and surgery-induced spatial reference learning and memory deficits.

Conclusions: MD2 contributes to the occurrence of PND by regulating α5GABA_ARs in aged mice, and Tat-CIRP-CMA is a promising neuroprotectant against PND.