Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Michigan 48824, USA.
Toxicol Sci. 2011 Nov;124(1):169-78. doi: 10.1093/toxsci/kfr213. Epub 2011 Aug 24.
Intranasal aspiration of satratoxin G (SG), a mycotoxin produced by the black mold Stachybotrys chartarum, selectively induces apoptosis in olfactory sensory neurons (OSNs) in mouse olfactory epithelium (OE) through unknown mechanisms. Here, we show a dose-dependent induction of apoptosis 24 h post-SG exposure in vitro as measured by increased activated caspases in the OP6 olfactory placodal cell line and increased propidium iodide staining in primary OE cell cultures. Intranasal aspiration of SG increased TUNEL (Terminal dUTP Nick End Labeling) staining in the neuronal layer of the OE and significantly increased the latency to find a buried food pellet, confirming that SG selectively induces neuronal apoptosis and demonstrating that SG impairs the sense of smell. Next, we investigated whether ATP can prevent SG-induced OE toxicity. ATP did not decrease apoptosis under physiological conditions but significantly reduced SG-induced OSN apoptosis in vivo and in vitro. Furthermore, purinergic receptor inhibition significantly increased apoptosis in OE primary cell culture and in vivo. These data indicate that ATP is neuroprotective against SG-induced OE toxicity. The number of cells that incorporated 5'-bromodeoxyuridine, a measure of proliferation, was significantly increased 3 and 6 days post-SG aspiration. Treatment with purinergic receptor antagonists significantly reduced SG-induced cell proliferation, whereas post-treatment with ATP significantly potentiated SG-induced cell proliferation. These data indicate that ATP is released and promotes cell proliferation via activation of purinergic receptors in SG-induced OE injury. Thus, the purinergic system is a therapeutic target to alleviate or restore the loss of OSNs.
鼻腔吸入由黑曲霉产生的真菌毒素 satratoxin G(SG)通过未知机制选择性诱导小鼠嗅上皮中的嗅觉感觉神经元(OSN)凋亡。在这里,我们通过体外 OP6 嗅基板细胞系中激活的半胱天冬酶的增加和原代 OE 细胞培养物中碘化丙啶染色的增加,显示出 SG 暴露后 24 小时的剂量依赖性凋亡诱导。SG 的鼻腔吸入增加了 OE 神经元层中的 TUNEL(末端 dUTP 缺口末端标记)染色,并显著增加了找到埋藏食物颗粒的潜伏期,证实 SG 选择性诱导神经元凋亡,并证明 SG 损害嗅觉。接下来,我们研究了 ATP 是否可以预防 SG 诱导的 OE 毒性。ATP 在生理条件下不会减少凋亡,但在体内和体外显着减少了 SG 诱导的 OSN 凋亡。此外,嘌呤能受体抑制在 OE 原代细胞培养物和体内显着增加了凋亡。这些数据表明 ATP 对 SG 诱导的 OE 毒性具有神经保护作用。掺入 5'-溴脱氧尿苷的细胞数量(增殖的量度)在 SG 吸入后 3 天和 6 天显著增加。嘌呤能受体拮抗剂的治疗显着降低了 SG 诱导的细胞增殖,而 ATP 的后期治疗显着增强了 SG 诱导的细胞增殖。这些数据表明,在 SG 诱导的 OE 损伤中,ATP 通过激活嘌呤能受体而被释放并促进细胞增殖。因此,嘌呤能系统是减轻或恢复 OSN 损失的治疗靶标。