Ginsenoside Re (GRe) exerts diverse effects. Based on our observations, the present study was designed to investigate GRe-exerted bidirectional regulation (BR) on the contractility of isolated jejunal segment. Six pairs of different low and high contractile states of rat jejunal segment were established and used in the study. Stimulatory effects on the contractility of jejunal segment were exerted by GRe (10.0 μM) in all 6 low contractile states, and inhibitory effects were exerted in all 6 high contractile states, indicating that GRe exerted BR on the contractility of jejunal segment. The effects of GRe on the phosphorylation of 20 kDa myosin light chain, protein contents of myosin light chain kinase (MLCK) and MLCK mRNA expression in jejunal segment in low and high contractile states were also bidirectional. GRe-exerted BR was abolished in the presence of neurotoxin tetrodotoxin or Ca2+ channel blocker verapamil or c-Kit receptor tyrosine kinase inhibitor imatinib. Atropine blocked the stimulatory effects of GRe on jejunal contractility in low-Ca2+-induced low contractile state; phentolamine, propranolol and l-NG-nitro-arginine blocked the inhibitory effects in high-Ca2+-induced high contractile state, respectively. In summary, GRe-exerted BR depends on jejunal contractile state and requires the presence of enteric nervous system, Ca2+, and interstitial cells of Cajal; the stimulatory effects of GRe on jejunal contractility are related to cholinergic stimulation and inhibitory effects are related to adrenergic activation and nitric oxide relaxing mechanisms.