Reprinted from the 1. 31. A. Archives of Pathology March 1958, f/of• 65, pp. 340-342

Copyright 1958. by American Medical Association

GLOMERULAR LESIONS PRODUCED IN THE RABBIT BY PREDNISONE AND PREDNISOLONE

SERGIO A. BENCOSME, M.D.; D. LAURENCE WILSON, M.D., and DAVID A. ROSEN, M.D., Kingston, Ont., Canada

After administration of cortisone to rabbits for short periods, kidney lesions with some morphologic features of diabetic gbmerulosclerosis have been described by Rich and his co-workers,2 by Bloodworth and Hamwi,3’4 and by Wilens and Stumpf.5 The histologic changes consist of striking glomerular capillary dilatation and nodular glomertilar lesions which stain positively with periodic acid-Schiff (PAS) stain, fat stains, and fibrin stains. Deeply staining amorphous casts are frequently noted in various portions of the renal tubular system. In the course of studies on the nature and significance of these lesions the effects of steroids other than cortisone on rabbit kidney morphology were investigated. This report deals with experiments carried out using the,1 synthetic steroids, prednisone and prednisolone)’ These substances are reported to have greater anti-inflammatory activity than cortisone, while exerting negligible effects on carbohydrate or electrolyte balance.6

Material and Methods

Twenty-four male New Zealand white rabbits were employed. Six were normal animals, and ten had been made diabetic six weeks previously with a single intravenous dose of alloxan monohydrate. The left kidney of each of these 16 animals was removed with use of pentobarbital (Nembutal) and ether anaesthesia to serve as control material. ____________

Submitted for publication May 28, 1957. From the Departments of Pathology, Medicine and Ophthalmology Queen’s University. This work was supported by National health Grants and by grants from the Canadian Life Insurance Officers’ Association. * The prednisone and prednisolone used in these studies were provided by J. W. Brisick, of the Schering Corporation Limited.

Results

The animals were maintained on Sherwood feed unsupplemented by cyanocohalamin (vitamin B12) or chlortetracycline (Aureomycin) and were provided with 1% sodium

chloride solution as drinking water. The diabetic group received 10 mg. of prednisone suspension intraperitoneally each day and the nondiabetic group, 5 mg. Wherever the condition of the animal permitted the steroid was continued for a period of 21 days.

A further group, of eight rabbits, received a daily a daily intraperitoneal injection of 5 mg. of prednisolone suspension. After 21 days the left kidney was removed, and the animals continued to be given 15 mg. of prednisone intraperitonelly each clay for a further 21 days. These animals were maihtained on a similar feed hut were provided with ordinary tap water ad libitum. In all groups, body weight, nonfasting blood glucose, and 24-hour urine glucose were measured twice weekly. Blood pressure was measured in the prednisone-treated nondiabetic group by auscultation over the aorta below a pediatric sphygmomanometer cuff. Kidney slices were fixed in formol and Zenkerformol, sectioned at 2.5u, and stained with hematoxylin, phloxine, and saffron.7 Additional sections were stained with PAS, oil red 0, masson richrome, and phosphotungstic acid and hema toxylin. Results

RESULTS

The blood sugar level in the diabetic rabbits ranged between 200 and 570 mg. per 100 cc. and was unaffected by prednisone administration. The blood sugar level of the nondiabetic animals receiving either prednisone or prednisone remained within normal limits. In all cases the body weight remained essentially unchanged throughout the experiment. The nondiabetic rabbits given Prednisone were found to have a mean maximum blood pressure increase of 31 mm. of mercury systolic and 33 mm. of mercury diastolic. All animals were noted to excrete

amber   urine   containing   a   large   quantity   of   sediment.     The   urine  was   not   found   to  react  with  benzidine.  

The   kidneys   removed   at   time   of   autopsy   presented   a   petechial   mottling   on   the  capsular  and  cut   surfaces  of   the  cortex;  no  other  gross  alteration  was  observed.  All  animals   which   had   received   prednisone   and/or   prednisolone   for   21   days   showed  characteristic   alterations   indistinguishable   from   those   observed   with   cortisone   by  ourselves  anti  others   2-­‐5     (Figs.  1  and  2).  Kidneys   from  the  diabetic  group  of  animals  presented,   in   addition,   evidence   of   acute   pyelonephritis   and   tubular-­‐cell   calcium  accumulation.  

While  the  effects  of  prednisolone  and  cortisone  were  similar,  those  of  prednisone  at  the   dosages   employed  were  more   numerous   and   severe.     Preexisting   diabetes   and  the   presence   of   hypertension   had   no   significant   influence   on   the   severity   or  frequency  of  lesions.  

Comment  and  Conclusions  

Our  own  histologic  studies  have  led  us  to  agree  with  those  authors  2,1,8    who  find  that  the  cortisone-­‐induced  lesions  of  the  rabbit  kidney  resemble  those  of  human  diabetic  glomerulosclerosis.    A  relationship  is  suggested  too  by  recent  studies  

 

 

 

of   Sommers   and  Haley,9  who  have  demonstrated   in   the   glomeruli   and  arterioles  of  nondiabetic  humans  and  animals  treated  with  cortisone  the  presence  of  an  abnormal  

substance   identical   in   ultraviolet   absorptive   properties   with   the   nodules   of   human  intercapillarv   glomerulosclerosis.   The   present   study   demonstrates   that   the,1    

substituted   steroids,   prednisone   and   prednisolone,   can   produce   similar   kidney  damage   to   that   observed   in   our   cortisone   studies.   It   is   particularly   significant   that  these  lesions  occur  in  nondiabetic  animals  and  are  not  influenced  by  alloxan  diabetes  or  experimental  hypertension.  

Department  of  Ophthalmology,  Queen’s  University  (Dr.  Rosen).  

REFERENCES  

1.  Kimmelstiel,  P.,  and  Vilson,  C.  :  Intercapillary  Lesions  in  Glomeruli  of  Kidney,  Am.  .1.  Path.  12:83-­‐89,  1936.  

2.   Rich,   A.   It.;   Berthrong,   M.,   and   Bennett.   I.   L.,   Jr.:   Effect   of   Cortisone   upon  Experimental   Cardiovascular   and   Renal   Lesions   Produced   by.     Anaphylactic  Hypersensitivite.  Boll.  Johns  Hop  kins  Hosp.  87:549-­‐567,  1950.  

3.   Bloodworth,   J.   M.   B.,   Jr.,   and   Hamwi,   G.   J.     Histopathologv   of   Experimental  Glomerular   Lesions   Simulating   Human   Diabetic     Glomerulosclerosis,   Am.   J.   Path.  31:167-­‐187,  1955.  

4.   Bloodworth,   J.   M.   B.,   Jr.,   and   Hamwi,   G.   J.   Experimental   Diabetic  Glomerotosclerosis,  Diabetes  5  :37-­‐43,  1956.  

5.  Wilens,  S.L.,  and  Stumpf,  H.  H.:  Nodular  and  Fatty  Glomerular  Lesions  in  Rabbits  on  Cortisone,  Am.  J.  Path.  31:275-­‐295,  1955.  

6.  Bunim,   J.   J.   ;  Pechet,  M.  M.,  and  Bollet,  A.   j.   :  Studies  on  Metacortandralone  and  metacortandralone   in   Rheumatoid   Arthritis:   Antirheomatic   Potency,   Metabolic  Effects,  and  Hormonal  Properties,  J.  A.  M.  A.  157:311-­‐318,  1955.  

7.  Bencosme,  S.  A.:  A  Trichrome  Staining  Method  for  Routine  Use,  Am.  J.  Clin.  Path.  24:  1324-­‐1328,  1954.  

8.  Becker,  B.;  Maengwyn-­‐Davies,  G.  D.;  Rosen,  D.;  Friedenwald,  J.  S.,  and  Winter,  F.  C.  :   The   Adrenal   Cortex   and   B-­‐Vitamins   in   Diabetic   Retinopathy,   Diabetes   3:175-­‐187,  1954.  

9.  Sommers,  S.,  and  Haley,  K.  H.:  Similarity  of  Glomerular  Ultraviolet  Absorptions   in  Diabetes  Mellitus  and  After  Cortisone  Therapy,  Proc.  Soc.  Exper.  Biol.  &  Med.  91:263-­‐265,  1956.