The La Liga leaders dominated the match at the Groupama stadium but lacked accuracy up front and were grateful to their goalkeeper Marc Andre ter Stegen who made two great saves.
Lyon were lively in the first half but ran out of steam after the break and were holding on in the closing stages as Lionel Messi tried in vain to open up the home defence.
Barca had a total of 25 shots but only five on target and their best chance came in the last minute when Sergio Busquets’s 18-metre attempt was tipped over the bar by Anthony Lopes.
The second leg will be at the Nou Camp on March 13, a game where Lyon will welcome back France striker and captain Nabil Fekir, who was suspended on Tuesday.
OL coach Bruno Genesio sprung a surprise by fielding Martin Terrier on the left wing instead of Maxwell Cornet and the 21-year-old did not disappoint.
Barca dominated possession early on, but Lyon had the first two shots on target and it took a couple of superb saves from Ter Stegen to deny the hosts, who never came that close again.
He stretched to his left to parry a vicious low shot by Hassem Aouar in the fifth minute before making a one-handed save to tip a 25-metre Terrier missile onto his crossbar.
Barca forward Ousmane Dembele came close in the 19th minute when he dribbled past Dubois on the left flank, only for keeper Anthony Lopes to deny him in a one-on-one.
The visitors had the upper hand but were uncharacteristically wasteful as they spurned several counter-attacking opportunities.
A Dembele attempt shaved Lopes’s post before Terrier skied his shot over the bar after a superb series of one-twos tore the Barca defence apart as both sides tried to break the deadlock.
Lyon were under siege early in the second period but their defence was perfectly marshalled by Jason Denayer with Dubois also proving decisive against Luis Suarez.
Lopes beat Messi’s angled shot away to keep his side level before Dembele was replaced by Philippe Coutinho, who had a clear chance in the 76th but was denied by in-form Lopes.
Barca have now failed to win any of their last six away games in the Champions League knockout stage, with their last victory coming at Arsenal in February 2016.
Solid and smart Bayern hold Liverpool at Anfield
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Liverpool's Naby Keita in action with Bayern Munich's James Rodriguez and Robert Lewandowski - Champions League - Round of 16 First Leg - Liverpool v Bayern Munich - Anfield, Liverpool, Britain - February 19, 2019. (Photo: Reuters)
The same day, Bayern Munich showed all their Champions League experience and quality organisation as they held Liverpool to a goalless draw in the first leg of their last 16 match at Anfield.
The result leaves it all to play for in the second leg in Munich on March 13, although the German side will be without defender Joshua Kimmich who will be suspended after picking up a yellow card.
There were few clear-cut chances in the game although Liverpool created several half-openings in the first period.
The first was one of the best they managed - Henderson found Salah, who had snuck in behind Niklas Sule, but the Egyptian’s toe-poke at full stretch was straight at Manuel Neuer.
At the other end, Liverpool goalkeeper Alisson Becker had to be alert to keep out a mis-hit clearance from Joel Matip that hit him in the chest.
Kingsley Coman fired into the side-netting after a sloppy give away from Alisson but that was a rare sign of jitters from either defence.
Bayern were content to sit deep after the break and the closest Klopp’s side came in the second half was a diving header from Sadio Mane in the 86th minute which Neuer pushed around the post.
