Objective To study the effect of prunella vulgaris on calcium ion channel TRPV5 in rat, and investigate its mechanism on calcium oxalate nephrolithiasis. Methods The Wistar rats were randomly divided into four groups(n=10 each): A group(normal control), B group(stone formation model), C group(treated by 50 g/L methanolic extract from prunella vulgaris), D group(treated by 100 g/L methanolic extract from prunella vulgaris). The model rats with renal calcium oxalate stone formation were induced by intragastric administration of ethylene glycol and ammonium chloride. After 4 weeks, the concentration of serum creatinine(Cr), blood urea nitrogen(BUN), Ca²⁺, P, 24 h urinary excretion Ca²⁺, Mg²⁺, and oxalate were detected. Renal samples were collected to prepare tissue slices and observe the pathological changes. The expression of TRPV5 was detected by Western blotting and real-time PCR. Results Compared with control group, nephrolithiasis rats in B group(model group)showed significant pathological changes in calcium oxalate calculus. After prunella vulgaris treatments, the renal pathological changes were shown. The concentration of Cr, BUN and urine calcium in group C and D were lower than those in group B(P<0.05), There was no difference in serum Ca²⁺,P, urine Mg²⁺, and oxalate between group C, D and group B(P>0.05). The expressions of TRPV5 protein and mRNA in group C and D were higher than those in group B(P<0.05). Conclusion The results showed that prunella vulgaris may increase the expression of TRPV5 in rats, while decrease the urine Ca²⁺, and reduce calcium oxalate saturation. It can inhibit renal calcium oxalate stone formation.