Seven unusual new ene-yne hydroquinones (<b>1</b>-<b>3</b>, <b>5</b>-<b>8</b>), including three rare glycosylated derivatives named pestalotioquinosides A-C (<b>6</b>-<b>8</b>), were obtained from the marine-derived strain SCSIO41403 of the fungus <i>Pestalotiopsis neglecta</i>. Their structures including absolute configurations were elucidated by spectroscopic analysis and induced electronic circular dichroism experiments. In silico molecular docking and <i>in vitro</i> surface plasmon resonance studies showed that pestalotioquinoside C (<b>8</b>) could act as a liver X receptor alpha (LXRα) modulator. Further study showed that LXR target gene <i>ABCA1</i> was significantly upregulated by <b>8</b>, which revealed <b>8</b> as a potential LXRα agonist.