title: [Effects of ammonium pyrrolidine dithiocarbamate (PDTC) on osteopontin expression and autophagy in tubular cells in streptozotocin-induced diabetic nephropathy rat].
abstract: Objective:To investigate the effects of ammonium pyrrolidine dithiocarbamate (PDTC) on tubulointerstitial inflammatory molecules and autophagy in diabetic nephropathy (DN) rats.Methods:Twenty-four male Sprague-Dawley rats were assigned to DN group (n=6) and DN+ PDTC group (n=6, PDTC, ip, 100 mg·kg-1·d-1), all received streptozotocin (STZ) 60 mg/kg intraperitoneally, and the other 12 rats were randomly divided into control group (n=6) and PDTC group (n=6). At the end of 12 weeks, after serum creatine (Scr) and 24-hour urinary protein were determined, rats were sacrificed to determined the renal pathological damages and the changes of nuclear factor (NF)-κB p65, p62, osteopontin (OPN), microtubule associated protein 1 light chain 3 (LC3)-Ⅱ/LC3-Ⅰ, nuclear p-NF-κB p65 by immunohistological stainning and Western blot, and ultrastructural changes of autophagic process was observed by electron microscopy (EM).Results:Scr was similar among the four groups (P>0.05). The levels of urinary protein in DN group and DN + PDTC group were significantly higher than the other two groups (allP<0.01), but the level of urinary protein in DN + PDTC group was lower than that of DN group (P<0.05). DN + PDTC group had less tubulointerstitial damage compared with DN group (P<0.05). Among the four groups, expressions of p62, p65, OPN of tubulointerstitial area in DN group were significantly higher than that of the other groups (allP<0.05), and Western blot showed that DN+ PDTC group had less expressions of NF-κB p65, nuclear p-p65, OPN and more expresssion of LC3-Ⅱ/LC3-Ⅰ compared with DN group (allP<0.05), which were consistent with the decreased autophagic vacuoles and increased mitochondria dysfunction revealed by EM. Correlation analysis showed that renal LC3-Ⅱ/LC3-Ⅰ was negatively correlated the expressions of nuclear p-p65 and OPN (r=-0.45,P=0.02;r=-0.50,P=0.01), and p62 was positively correlated the expressions of nuclear p-p65 and OPN (r=0.33,P=0.01;r=0.41,P=0.01).Conclusion:Tubular NF-κB activation is closely related to autophagy dysfunction in DN rats, and PDTC may enhance autophagy activity in tubule cells by blocking NF-κB activity.
authors.affiliation: *Department of Nephrology, the General Hospital of Tianjin Medical University, Tianjin 300052, China.
authors.forename: S
authors.initials: S
authors.lastname: Gao
authors.forename: J Y
authors.initials: JY
authors.lastname: Jia
authors.forename: T K
authors.initials: TK
authors.lastname: Yan
authors.forename: Y M
authors.initials: YM
authors.lastname: Yu
authors.forename: W Y
authors.initials: WY
authors.lastname: Shang
authors.forename: L
authors.initials: L
authors.lastname: Wei
authors.forename: Z F
authors.initials: ZF
authors.lastname: Zheng
authors.forename: P
authors.initials: P
authors.lastname: Fang
authors.forename: B C
authors.initials: BC
authors.lastname: Chang
authors.forename: S
authors.initials: S
authors.lastname: Lin
mesh_terms: D000818:Animals; D001343:Autophagy; D015153:Blotting, Western; D003928:Diabetic Nephropathies; D007668:Kidney; D008297:Male; D016328:NF-kappa B; D053495:Osteopontin / Q000378:metabolism*; D011759:Pyrrolidines / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013311:Streptozocin; D013859:Thiocarbamates / Q000627:therapeutic use*
chemical_list: D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; D053495:Osteopontin; C020972:pyrrolidine dithiocarbamic acid; D013311:Streptozocin