A late strike from Taylor Perry ensured Wolves got a first win of the season against a battling nine-man Rovers at Leyland.
After Matty Platt and then Charley Doyle were handed each handed an early bath, Perry crashed home in the final minute of normal time.
The Rovers pair saw red just minutes after Harry Chapman has levelled the score for thehosts in a fiery second half at the Lancashire FA HQ.
After a goalless first half, the game sprung into life with Meritan Shabani's opener, prior to Chapman's pinpoint finish for the equaliser.
Barr handed outings to first team trio Sam Hart, Chapman and Dominic Samuel, who all gave themselves a fitness boost with game time
The visitors had the first opportunity of the game inside the opening 10 minutes, with Benny Ashley-Seal firing wide of Joe Hilton's right-hand post from just inside the box.
At the other end, the returning Samuel smashed an effort from range that flew past the upright in a lively start at Leyland.
The visitors almost took the lead through Shabani, but the midfielder fired wide of the target 10 minutes before the break.
The best chance for Rovers came from rampaging left back Lewis Thompson, but the youngster's excellent solo run ended in a cross-shot that just evaded Stefan Mols at the back post.
But just five minutes after the interval the visitors grabbed the advantage, with Shabani in the right place at the right time to turn home a cross from John Kitolano.
But Rovers bounced back almost instantly, with Chapman rifling into the bottom corner from the edge of the box to equalise.
However, just seconds later and Barr's boys were reduced to 10 men when Platt was shown a straight red card for a cynical challenge on Kitolano in the middle of the park.
The cause wasn't helped just moments later when Doyle was shown a second yellow for a late challenge, leaving Rovers to play the final 23 minutes with nine men.
Wolves were pushing up searching for a winner, with Lewis Richards seeing Hart in the way of his goalbound strike, before a brilliant block from Tom White denied Elliot Watt from inside.
Rovers were defending well and putting their bodies on the line, with Watt slicing over with just a couple of minutes remaining.
But just as it looked like the home side would bravely hold on to a battling point, Perry smashed into the bottom corner in the final minute of normal time to break Rovers' hearts.
Rovers are next in action on Friday 30th August at Brighton & Hove Albion, kick-off 7pm.
Rovers U23s: Hilton, Doyle, Thompson, White, Platt (c), Grayson, Mols, Lyons, Samuel (Brennan, 46 (Barnes, 68), Chapman (Annesley, 82), Hart.
Unused substitutes: Eastham, Jackson.
Goal: Chapman, 58
Booked: Doyle, 32, 67, Mols, 90
Sent off: Platt, 60, Doyle, 67
Wolves Under-23s: Sondergaard, Buur (Thompson, 77), Nya (Cristovao, 56), Francis, Kitolano, Perry, Watt, Ashley-Seal, Jordao (Diallo, 46), Shabani.
Unused substitutes: Dai, Samuels.
Goal: Shabani, 51
Booked: Nya, 16, Perry, 71, Richards, 90.