Abstract: Objective To access the progressive,permanent,and irreversible damage of kidney of rabbit for repeat ESWLs,seek digital times-effect relationship of ESWL,learn the processes of eternal and irreversible damages of renal function caused by multiple and repeat ESWLs,and guide ESWL for the patient with recurrent or complicated kidney stone to reduce renal damage and prevent severe renal function damage. Methods Animal and design of the studies are same as study I.We killed the rabbits,removed the kidney,and performed historical examinations our earlier after ESWL,observed changes of histological structure and fibrosis of glomeruli and tubules;digitized the numbers of glomeruli under microscope by H.E.and Masson stain;detected expression of TGF-β₁,collagen Ⅳ by immunohistochemical stain under microscope. Results ①Accumulated damage of kidney occurred after repeat ESWL with structural changes of nephron,and fibrolification of glomeruli and tubules occurred gradually with the increasing of ESWL times.②There are significant difference on average numbers of glomerular under 100 times microscope fields between ESWL group and control group.After 6-times of ESWL,numbers of glomeruli decreased significantly,and degree of renal fibrosis gradually enhanced with increased ESWL times.③There were significant differences in comparison of degree of TGF-β₁ and Collagen Ⅳ expression increased,and the degree markedly enhanced after 4 times of ESWL,and as the times of ESWL increasing,the degree increased gradually. Conclusion Multiple,repeat ESWLs can cause accumulated damage of kidney with fibrosis;After 4-times ESWLs,expressions of TGF-β₁ and Collagen Ⅳ increased,these predicted severe fibrilification would occur,which can cause permanent,irreversible kidney damage.ESWL should be performed with strict indications for renal stone patients and control ESWL within 4 times to prevent the irreversible accumulated damage of kidney and protect renal function maximum.