Shen Shuai II recipe improves renal hypoxia to attenuate renal injury in 5/6 renal ablation/infarction rats and effect evaluation using blood oxygenation level-dependent functional magnetic resonance imaging

Figures & data

Table 1. Composition and information of SSR.

Herb	Chinese name	Medicinal part	Quantities (g)
Codonopsis Radix	Dang shen	Rhizome	15
Epimedii Folium	Yin yang huo	Leaf	15
Salviae Miltiorrhizae Radix et Rhizoma	Dan shen	Root and rhizome	15
Angelicae Sinensis Radix	Dang gui	Root	15
Prepared Rhei Radix et Rhizoma	Da huang	Root and rhizome	15
Perillae Folium	Zi su ye	Leaf	15
Chuanxiong Rhizoma	Chuan xiong	Rhizome	15
Persicae Semen	Tao ren	Seed	15
Coptidis Rhizoma	Huang lian	Rhizome	6

Figure 1. Construction of the 5/6 (A/I) rat model of CKD. 5/6 (A/I), 5/6 renal ablation/infarction; CKD, chronic kidney disease.

Figure 2. Flowchart of BOLD-fMRI examination in rats. BOLD-fMRI, blood oxygenation level-dependent functional magnetic resonance imaging.

Figure 3. BOLD-fMRI T2*WI and T2* map in the coronal plane of rats. (A, B) The rat from Sham group. (C, D) The rat from 5/6 (A/I) group. (E, F) The rat from 5/6 (A/I) + LOS group. (G, H) The rat from 5/6 (A/I) + SSR group. ROI-based technique, with placement of circled ROIs in the renal cortex and medulla. BOLD-fMRI, blood oxygenation level-dependent functional magnetic resonance imaging; ROI, region of interest.

Figure 4. Bland-Altman plots of BOLD-fMRI analyses between observers. BOLD-fMRI, blood oxygenation level-dependent functional magnetic resonance imaging.

Figure 5. Effects of SSR on renal oxygenation in 5/6 (A/I) rats. (A) The level of COT2* was evaluated by BOLD-fMRI after treatment. (B) The level of MET2* was evaluated by BOLD-fMRI after treatment. (C) The expression of HIF-1α protein was determined by western blot. (D) Quantitative analysis of HIF-1α protein level (n = 4). Data were analyzed by One-way ANOVA. Values are mean ± SE. *p < 0.05; **p < 0.01; ***p < 0.001; ^****p < 0.0001. COT2*, cortical T2*; MET2*, medullary T2*; LOS, Losartan; SSR, Shen Shuai II Recipe.

Table 2. Comparisons of COT2* and MET2* between four groups. ($\overline{x}\pm s$).

Class	Sham (n = 6)	5/6 (A/I) (n = 8)	5/6 (A/I) + LOS (n = 8)	5/6 (A/I) + SSR (n = 9)	F	p value
COT2*	54.43 ± 4.37	33.11 ± 4.95	42.60 ± 7.43	55.39 ± 5.89	25.521	< 0.0001
MET2*	35.94 ± 2.20	21.95 ± 4.28	27.15 ± 5.98	35.99 ± 3.92	19.160	< 0.0001
t	8.658	9.568	10.921	9.605	─	─
p value	0.0003	< 0.0001	< 0.0001	< 0.0001	─	─

COT2*, cortical T2*; MET2*, medullary T2*; LOS, Losartan; SSR, Shen Shuai II Recipe.

Figure 6. Effects of SSR on kidney and liver function and proteinuria in 5/6 (A/I) rats. The levels of SCr (A) and BUN (B) before treatment. The levels of SCr (C), BUN (D), CysC (E), UA (F), Ccr (G), 24-h Upr (H), UACR (I), ALT and AST (J) were measured after 8-week treatment. Data were analyzed by One-way ANOVA or Student’s t-test. Values are mean ± SE. *p < 0.05; **p < 0.01; ***p < 0.001; ^****p < 0.0001. SCr, serum creatinine; BUN, blood urea nitrogen; CysC, serum cystatin C; UA, serum uric acid; Ccr, creatinine clearance rate; Upr, urinary protein; UACR, urinary albumin:creatinine ratio; ALT, alanine aminotransferase; AST, aspartate aminotransferase.

Figure 7. Effects of SSR on renal injury and fibrosis in 5/6 (A/I) rats. (A) The expression of FN, Col-I, and α-SMA proteins were determined by western blot. (B) Quantitative analysis of FN, Col-I, and α-SMA levels (n = 4). (C) Representative photomicrographs of HE and Masson’s trichrome staining. 200 × magnification. (D) The severity of tubular damage was assessed by the tubular injury score (n = 8). (E) Semiquantitative analysis of collagen area (n = 4). Data were analyzed by One-way ANOVA. Values are mean ± SE. *p < 0.05; **p < 0.01; ***p < 0.001 vs. Sham group, ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. 5/6 (A/I) group. LOS, Losartan; SSR, Shen Shuai II Recipe.

Figure 8. Correlation of COT2* and MET2* with SCr (A), BUN (B), CysC (C), UA (D), Ccr (E), Hemoglobin (F), 24-h Upr (G), and UACR (H). Data were analyzed by Pearson’s correlation coefficient. COT2*, cortical T2*; MET2*, medullary T2*; SCr, serum creatinine; BUN, blood urea nitrogen; Cys C, serum cystatin C; UA, serum uric acid; Ccr, creatinine clearance rate; Upr, urinary protein; UACR, urinary albumin:creatinine ratio.

Availability of data and materials

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.