BACKGROUND It is considered that the pathogenesis is closely related to an excessive production of pro-inflammatory cytokines caused by bacterial toxins and an imbalance between pro-inflammatory and anti-inflammatory mediators. METHODS This work investigates the effect of electro-acupuncturing (EA), at Zusanli point (ST36) on plasma cytokine release and organ dysfunction and their mechanism in conscious rats with endotoxin challenge. RESULTS EA at Zusanli points obviously lowered the elevated levels of plasma TNF-α, and attenuated changes in parameters relevant to various organ functions at 2 h after LPS challenge. α-BGT injection or bilateral cervical vagotomy could weaken or eliminate the effects of EA, and further aggravated the elevated levels of pro-inflammatory cytokines and organ dysfunction. CONCLUSIONS The results suggested that EA at Zusanli points significantly reduced the release of pro-inflammatory cytokines and organ dysfunction after LPS challenge by activating cholinergic anti-inflammatory pathway.