Fig 1.
Schedule of the accelerated nephrotoxic serum nephritis model.
Mice were divided into six groups (n = 6): normal (N), NTS control (NTS) saikosaponin C (SSC) control (SC), SSC 5 (L), 10 (M), 20 (H) mg/kg/day. Mice in groups NTS, L, M, and H were pre-immunized by injection with rabbit IgG then were induced with 200 μL NTS intravenous injection, while mice in groups N and SC were injected with normal saline. On the next day, mice in groups SC, L, M, and H were administered SCC once daily for 14 days. The urine was collected at four intervals for analysis of urine protein, methylglyoxal and D-lactate. The serum was collected for biochemical assays as serum creatinine and BUN. The kidneys were collected for histological examination (hematoxylin and eosin, Masson’s trichrome, and sirius red stain), immunoblotting analysis of glyoxalase 1 (GLO 1), and immunohistochemistry analysis of F4/80.
Fig 2.
Urinary protein levels of each experimental group.
N: normal group, SC: saikosaponin C control group, NTS: nephrotoxic serum control group, L: SSC 5 mg/kg/day treatment group, M: SSC 10 mg/kg/day treatment group, H: SSC 20 mg/kg/day treatment group. ★p<0.05 compared to N group, #p<0.05 compared to NTS control group.
Table 1.
BUN and serum creatinine (SCr) levels of each experimental group.
Fig 3.
Hematoxylin and eosin stained renal tissue from different experimental groups (×200 magnification).
(A) Normal, (B) SSC control, (C) NTS control (arrow: the crescent), (D) NTS + SSC 5 mg/kg, (E) NTS + SSC 10 mg/kg, and (F) NTS + SSC 20 mg/kg. (G) Histological scores for the experimental groups. ★p<0.05 compared to N group, #p<0.05 compared to NTS control group.
Fig 4.
Masson’s trichrome-stained renal tissues from different experimental groups (×200 magnification).
(A) Normal, (B) SSC control, (C) NTS control (arrow, the part of fibrosis), (D) NTS + SSC 5 mg/kg, (E) NTS + SSC 10 mg/kg, and (F) NTS + SSC 20 mg/kg. (G) Quantification of the area of fibrosis for each experimental group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 5.
Sirius red-stained renal tissues from different experimental groups (×200 magnification).
(A) Normal, (B) SSC control, (C) NTS control (arrow, the part of fibrosis), (D) NTS + SSC 5 mg/kg, (E) NTS + SSC 10 mg/kg, and (F) NTS + SSC 20 mg/kg. (G) Quantification of the area of fibrosis for each experimental group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 6.
Immunohistochemistry stained renal tissues for F4/80 from different experimental groups (×200 magnification).
(A) Normal, (B) SSC control, (C) NTS control (arrow, the part of macrophage infiltration), (D) NTS + SSC 5 mg/kg, (E) NTS + SSC 10 mg/kg, and (F) NTS + SSC 20 mg/kg. (G) Quantification of the area of fibrosis for each experimental group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 7.
Renal TNF-α levels in the different experimental groups.
N: normal group, SC: saikosaponin C control group, NTS: nephrotoxic serum control group, L: SSC 5 mg/kg/day treatment group, M: SSC 10 mg/kg/day treatment group, H: SSC 20 mg/kg/day treatment group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 8.
Urinary methylglyoxal levels in the different experimental groups.
N: normal group, SC: saikosaponin C control group, NTS: nephrotoxic serum control group, L: SSC 5 mg/kg/day treatment group, M: SSC 10 mg/kg/day treatment group, H: SSC 20 mg/kg/day treatment group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 9.
Urinary D-lactate levels in different experimental groups.
N: normal group, SC: saikosaponin C control group, NTS: nephrotoxic serum control group, L: SSC 5 mg/kg/day treatment group, M: SSC 10 mg/kg/day treatment group, H: SSC 20 mg/kg/day treatment group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 10.
Immunoblotting analysis of glyoxalase 1 in renal homogenates from different experimental groups on day 15.
N: normal group, SC: saikosaponin C control group, NTS: nephrotoxic serum control group, L: SSC 5 mg/kg/day treatment group, M: SSC 10 mg/kg/day treatment group, H: SSC 20 mg/kg/day treatment group. ★p<0.05 compared to normal group, #p<0.05 compared to NTS control group.
Fig 11.
The relationship between methylglyoxal and D-lactate in saikosaponin C treatment of nephrotoxic serum nephritis.
Nephrotoxic serum nephritis induced inflammation up-regulated glycolysis then cause methylglyoxal and D-lactate increased in the urine. The appropriate dosage of saikosaponin C treatment may improve the immune-reaction and down-regulate methylglyoxal and D-lactate production.