The role of the molecular alignment distribution with high harmonic generation (HHG) in aligned CO2 molecules is investigated and discussed in this letter. We first present the molecular field-free alignment and the polar plot of the weighted alignment distribution, and then also give the angular distribution of the molecular HHG. At last, we theoretically and experimentally demonstrate the alignment dependence for the molecular HHG. These results systematically verify the role of the molecular alignment distribution in the HHG process, which is useful for investigating the molecular dynamics in practical application.