The Lions are now pointless from their first two matches.
Joint-manager Marcus Bignot said: “We had chances and it was a game that could have gone our way, we have to focus on Saturday now.”
It was a case of differing results from their opening matches, with Guiseley looking for their first points after losing at Boston United, while Chorley were aiming to build off a win against York City. The hosts had the first chance when a Scott Garner header caused problems for Chorley but they were just about able to clear it away from danger.
The first opportunity for the visitors fell to Marcus Carver who saw his effort go over the crossbar, after he was found in the area by Alex Newby.
The visitors began to grow into the game and went ahead when Newby collected a pass and on the angle was able to squeeze his effort past Guiseley keeper Joe Green.
A scrappy opening to the second period saw Carver hit a shot across the face of the goal and wide, with the visitors looking to double their advantage.
The Lions thought they had levelled when Niall Heaton headed in, but the linesman’s flag was up for offside and it was disallowed.
Just when it looked like Guiseley were taking control of the game, Chorley broke and doubled their lead.
A Matt Challoner effort hit the crossbar and it looked like the danger was cleared but Adam Blakeman hit a fine effort from 20 yards to seal the win for Chorley.
Guiseley are back at home on Saturday as they look to pick up their first points of the season against AFC Telford United.
In the National League, promoted Harrogate Town picked up their second point at Hartlepool United, following a second succesive 2-2 draw.
Callum Howe was the Town hero scoring a 90th-minute leveller at Victoria Park.
All the goals came in the second half, with Niko Muir (53) opening the scoring for Pools, Dom Knowles replying (60). Former Guisley man Jake Cassidy (76) restored the hosts’ lead before Howe’s heroics.