Vitamin B₁₂ supplement alleviates N′-nitrosodimethylamine-induced hepatic fibrosis in rats

Figures & data

Figure 1. Liver glycogen reserves in experimental animals during NDMA-induced hepatic fibrosis and subsequent treatment with vitamin B₁₂. Values are expressed as mean ± SD (*p < 0.05, **p < 0.001).

Table 1. Changes in biochemical parameters of hepatic injury in the sera and liver of control and treated rats.

Biochemical parameters	Control [A]	NDMA-treated [B]	Percent change [B−A/B × 100]	Vitamin B12-supplemented [C]	Percent change [B−C/B × 100]
SALP (U/L)	316 ± 22.2	694 ± 29.09*	54.46 ↑	355 ± 19.86*	48.84 ↓
SGOT (U/L)	125 ± 13.5	180 ± 15.21*	30.55 ↑	135 ± 22.9*	25.00 ↓
SGPT (U/L)	55 ± 3.9	67 ± 4.16*	17.91 ↑	59 ± 8.2*	11.94 ↓
TBil (mg/dL)	0.314 ± 0.03	1.35 ± 0.13**	76.74 ↑	0.44 ± 0.06*	67.40 ↓
Hydroxyproline (µg/mL serum)	10.72 ± 0.99	14.01 ± 1.21**	23.48 ↑	11.34 ± 1.1*	19.05 ↓
Hydroxyproline (µg/mL liver extract)	0.147	0.505**	70.89↑	0.205**	59.40↓
DBIL (mg/dL)	0.15 ± 0.011	0.165 ± 0.04	9.09 ↑	0.151 ± 0.02	8.48 ↓
TSP (g/dL)	7.4 ± 0.44	6.74 ± 1.01	10.31 ↓	7.15 ± 0.77	6.08 ↑

SALP, sera alkaline phosphatase; SGOT, sera glutamic oxaloacetic transaminase; SGPT, sera glutamic pyruvic transaminase; Tbil, total bilirubin; DBIL, direct bilirubin; TSP, total sera proteins; ↑, increase; ↓, decrease.

Treatment with N′-nitrosodimethylamine (NDMA) in doses of 10 mg kg⁻¹ body weight was given to animals for 21 days under laboratory conditions. Another treatment group was supplemented with vitamin B₁₂. (Values are expressed as mean ± SD.)

*p < 0.05, **p < 0.001.

Figure 2. Vitamin B₁₂ levels in the liver and plasma of experimental animals during NDMA-induced hepatic fibrosis and after treatment with vitamin B₁₂. Values are expressed as mean ± SD (*p < 0.05, **p < 0.001).

Figure 3. Hematoxylin and eosin (H & E) staining of rat liver during NDMA-induced hepatic fibrosis and subsequent treatment with vitamin B₁₂. (A) Control liver showing central vein and characteristic lobular architecture (10 × ). (B) Day 7. Peripheral lymphocyte infiltration (4×). (C) Day 14. Demonstrating Kupffer cell hyperplasia (40×). (D) Day 21. Liver congestion, lymphocyte infiltration and well developed fibrosis (10×). (E) Control liver with normal structure subsequent to vitamin B₁₂ treatment only (4×). (F) Day 7. Liver showing lymphocyte and neutrophilic infiltration with mild fibrosis (40×). (G) Day 14. Inflammation absent with decreased fibrosis (10×). (H) Day 21. Restoration of normal liver architecture with increasing density of regenerating hepatocytes (10×).

Figure 4. Immunohistochemical staining for α-SMA during the pathogenesis of NDMA-induced hepatic fibrosis and subsequent treatment with vitamin B₁₂. (A) Normal liver with central vein (4×). (B) Day 7. Focal α-SMA staining around central vein positively stained stellate cells (10×). (C) Day 14. Focal positivity and α-SMA stained regions showing widespread activation of hepatic stellate cells (40×). (D) Day 21. Stellate cells showing intense focal staining of α-SMA in fibrotic region (40×). (E) Normal liver architecture in vitamin B₁₂ treated rats (4×). (F) Day 7. Focal positivity of α-SMA (4×). (G) Day 14. Scattered and decreased focal positive staining of α-SMA showing reduced number of activated hepatic stellate cells (10×). (H) Day 21. Regenerating hepatocytes with scant positivity around central vein (4×).