Abstract Background: Chronic hyperglycemia is linked to either subfertility or infertilityamongdiabeticmales.Pioglitazone isoneof thethiazolidinediones (TZDs) drugs that are selective peroxisome proliferator‐activated receptor (PPAR‐γagonistsusedfortreatingtype2diabetesmellitus(T2DM). Aim:ThisstudyaimstoexplorethepossibleeffectoflowPioglitazonedoseand omega(ω‐3)onratmalereproductivefunction.Furthermore,weevaluatedthe add‐on effect of combined use of lowPioglitazone dose of and ω‐3 on reproductivefunctionsinadultmaleT2DMrats. Methods:Fiftyadultmaleratswereincludedandsubdividedintocontrolandfour testsubgroups.T2DMwasinducedintestgroupsandsubdividedintonon‐treated T2DM,ω‐3treated, 0.6mg/kgPioglitazone treated, andcombinedtreatedgroup (orallybygavage).Following16weeks,finalbodyweight,testicularweight,fasting plasmaglucose, andserumtestosterone levelsweremeasured. Semenanalysis, testicular testosterone, malondialdehyde (MDA) concentrations, superoxide dismutase (SOD) activity, immunohistochemistry staining for apoptosismarker B‐celllymphomaprotein2(Bcl‐2),proliferationmarkerasproliferatingcellnuclear antigen (PCNA), estrogen receptor α (ERα), androgen receptor (AR) were determined. Caspase‐3, nuclear factor‐kappaB (NF‐kB), glucose transporter 3 (GLUT3), 17β‐hydroxysteroid dehydrogenases (17β‐HSD) PPARγ, and PPARα genesexpressionwereanalyzedbyreal‐timepolymerasechainreaction(RT‐PCR). Results:OurfindingsrevealedthattreatmentwithlowdoseofPioglitazoneor ω‐3significantlyloweredfastingplasmaglucoseandMDAlevels,ameliorated diabeteseffectsonhistologicaldamage, improvedantioxidant activity(SOD), significantlyimprovedanti‐apoptosisBCL‐2andproliferation(PCNA),remarkably elevated ERα, AR, 17β‐HSD PPARγ, and PPARα expression with significant reduction in caspase‐3, NF‐kB genes expression and improved semenqualityaswell.Combineduseoflowdoseofandω‐3hasbettereffectson allmeasuredparameters.