The Bile Acid receptor TGR5 is well known to activate the cAMP pathway leading to CFTR activation and Cl- ions secretion, needed for bile alkalinization and hydration. However, during cystic fibrosis development, only 10 to 15 % of the patients present liver defects due to bile duct disorders, meaning that another process should compensate for the loss of CFTR activity. Interestingly, some bile acids had also been reported to mobilize Ca2+ ions in cholangiocytes. Using normal human cholangiocytes and cholangiocarcinoma cell lines, we confirmed by using a specific agonist, that TGR5 stimulation induced a Ca2+ release from the endoplasmic reticulum and an influx of extracellular Ca2+ ions. Next, this Ca2+ mobilisation allows an ATP (and UTP) release, leading to the activation of P2Y receptors, reinforcing this Ca2+ mobilisation. This study shows that activation of the BA receptor TGR5 has the capacity to induce the two main intracellular pathways, cAMP and IP3-Ca2+ in cholangiocytes. From our data, we speculate that the pathway we described will allow activation of the Ca2+-activated Cl- channels TMEM16A, to compensate in part or in totality the loss of CFTR in CF patients.