Shenhua Tablet inhibits mesangial cell proliferation in rats with chronic anti-Thy-1 nephritis

Abstract

Background: In China, mesangial proliferative glomerulonephritis (MsPGN) is one of the most common kidney diseases. In this study, we treated a rat model of chronic anti-Thy-1 MsPGN with Shenhua Tablet and evaluated whether the tablet was able to protect the kidney function. Thirty-six Wistar rats were randomly divided into six groups: (1) Sham surgery (Sham); (2) anti-Thy-1 nephritis model (Thy-1); (3) anti-Thy-1 nephritis model + irbesartan-treated (Irb); (4) anti-Thy-1 nephritis model + low-dose of Shenhua Tablet (SHL); (5) anti-Thy-1 nephritis model + medium-dose of Shenhua Tablet (SHM); (6) anti-Thy-1 nephritis model + high-dose of Shenhua Tablet (SHH).

Results: Thirteen weeks after drug treatment, urinary proteins were quantified and renal pathological changes were thoroughly examined at the time point of 24 h. Meanwhile, the expression levels of p-Erk1/2, cyclin D1 and p21 at the renal cortex were also tested. The levels of urinary proteins and total cholesterol in the blood were significantly reduced in rats treated with any drug tested in this study. The level of triglyceride was significantly reduced in all three Shenhua Tablet-treated groups. Renal pathomorphological scores were significantly improved in groups of Irb, SHM and SHH. Mesangial cell proliferation was significantly inhibited in any drug-treated group. p-Erk1/2 and cyclin D1 were downregulated whereas p21 was upregulated in the renal cortex.

Conclusions: Our study indicated that Shenhua Tablet is able to inhibit the abnormal proliferation of mesangial cells and to prevent kidney damage, which is likely associated with downregulation of p-Erk1/2 and reduced activity of its downstream target-cyclin D1.

Keywords: Chronic anti-Thy-1 glomerulonephritis; Erk; Mesangial cell proliferation; Shenhua Tablet.

Fig. 1

The levels of urinary protein in 24 h were monitored for each group over the experimental process (week 1, 3, 5, 7, 11 and 20 after injection of Thy-1 antibody). At the last time point W20, the urinary proteins in 24 h in all drug-treated groups were significantly lower (p < 0.01) than that in the Thy-1 group. Values were expressed as mean ± SD. * p< 0.05, ** p< 0.01 vs. Thy-1; ^# p< 0.05, ^## p< 0.01 vs. Irb group

Fig. 2

The levels of serum creatinine in each groups at different time points. The levels of serum albumin were quantified in all six groups at different time points (week 1, 3, 5, 7 and 20 after injection of Thy-1 antibody). Values were expressed as mean ± SD. * p< 0.05, ** p< 0.01 vs. Thy-1

Fig. 3

The effect of Shenhua Tablet on the biochemical indices in blood. a The levels of serum albumin were quantified in all six groups at different time points (week 1, 3, 5, 7 and 20 after injection of Thy-1 antibody). b The levels of total cholesterol in blood were quantified in all six groups at different time points mentioned above. c The levels of triglyceride were quantified in all six groups at different time points mentioned above. Values were expressed as mean ± SD. * p< 0.05, ** p< 0.01 vs. Thy-1. n = 6

Fig. 4

Shenhua Tablet inhibits proliferation of mesangial cells in rats with anti-Thy-1 nephritis and improves renal pathological scores. a Histological changes in PAS-stained sections at W20 (×400 magnification). Scale bar 50 µm. b The total number of glomerular cross section in the six groups. c PCNA-positive cells in the groups at W20. Scale bar 50 µm. d The PCNA labeling index in all six groups. The index was calculated according to the ratio of PCNA positive cells to total glomerular cells. e Histological scores of the renal lesions in all six groups. Values were expressed as mean ± SD. * p< 0.05, ** p< 0.01 vs. Thy-1

Fig. 5

Shenhua Tablet downregulates p-Erk1/2 and cyclin D1 while upregulating p21 in the renal cortex of rats with anti-Thy-1 nephritis. a Western blot was performed to detect the expression levels of p-Erk1/2, cyclin D1 and p21 in all drug-treated groups as well as the controls. b Quantification of the cyclin D1, p21 and p-Erk1/2. Expression level of p-Erk1/2 was normalized with total Erk1/2 while cyclin D1 and p21 were normalized with beta-actin. Each bar represents the mean ± SD of six rats. * p< 0.05, ** p< 0.01 vs. Thy-1; ^# p< 0.05, ^## p< 0.01 vs. Irb group

Fig. 6

A scheme of drug treatment for the animal model of chronic anti-Thy-1 MsPGN. Thirty-six Wistar rats were randomly divided into six groups: (1) Sham surgery group (Sham); (2) anti-Thy-1 nephritis model group (Thy-1); (3) anti-Thy-1 nephritis model + irbesartan-treated group (Irb); (4) anti-Thy-1 nephritis model + low-dose of Shenhua Tablet group (SHL); (5) anti-Thy-1 nephritis model + medium-dose of Shenhua Tablet group (SHM); (6) anti-Thy-1 nephritis model + high-dose of Shenhua Tablet group (SHH)