Figure 6. Paneth cell granule depletion with dithizone treatment.

A. Representative H&E (of 6 experiments, magnification 400X) of ileum from mice treated with vehicle (Li₂CO₃) or with dithizone 6 hrs prior. Note complete depletion of Paneth cell granules (arrows) after dithizone treatment (*). B. Representative (of 5 experiments, 400X) ileum lysozyme immunostaining. Note heavy lysozyme stain in Paneth cells (arrow heads) of mice treated with vehicle (Li₂CO₃). Paneth cell depletion with dithizone treatment decreased lysozyme staining in Paneth cells. In addition, lysozyme staining in Paneth cells was reduced after bilateral nephrectomy (BNx) compared to sham operated mice (Sham) and we were able to detect lysozyme staining in crypt lumen (arrows and enlarged insert of 2000X magnification). Villous lysozyme staining was also evident (*). C. Representative (of 3 experiments, 200X magnification and enlarged insert, 600X magnification) immunofluorescence stain for IL-17A (green) and CD68 (red) in small intestine of mice treated with dithizone (100 mg/kg, i.v. 6 hrs prior to renal ischemia or nephrectomy) and subjected to sham-operation, BNx or renal ischemia reperfusion (RIR). Paneth cell granule depletion reduces crypt IL-17A production after AKI. D. Dithizone treatment reduces plasma IL-17A levels (analyzed 5 hrs after acute kidney injury) in mice subjected to acute kidney injury (BNx or 30 min. RIR, N=4 each). Dithizone treatment also reduced IL-17A protein upregulation in small intestine (N=4) and in isolated crypts (N=4) 5 hrs after acute kidney injury. *P<0.05 vs. sham-operated mice. #P<0.05 vs. mice subjected to vehicle treated animals subjected to acute kidney injury. Error bars represent 1 SEM. E. Paneth cell granule depletion with dithizone treatment protects against hepatic injury after ischemic acute kidney injury (RIR) or bilateral nephrectomy (BNx). Mice were subjected to sham-operation (vehicle (veh) or dithizone Sham, N=4), BNx (N=6) or 30 min. RIR (N=6). Plasma was collected 5 hrs after BNx and 24 hrs after RIR. F. Paneth cell granule depletion with dithizone treatment also protects against acute kidney injury (creatinine and blood urea nitrogen) after RIR.