Effect of hepatic iron concentration reduction on hepatic fibrosis and damage in rats with cholestatic liver disease

Table 1 Weights and phlebotomy volume in different groups of rats (mean±SE)

Subgroup	n	Weight gain(% from baseline)	Mean phlebotomy Volume1(mL)	Liver weight(% of total body weight)	Splenic weight(% of total body weight)
Sham only	5	15.03  ±  18.70a	0  ±  0	3.97  ±  0.16a	0.41 ± 0.07a
Sham and phlebotomy later	4	9.4  ±  7.0	0.70  ±  0.13	3.62  ±  0.32	0.37 ± 0.04
Phlebotomy and sham later	6	-5.7 ± 17.3	0.86  ±  0.18	3.47  ±  0.19	0.38 ± 0.03
BDL only	9	-4.99  ±  3.90	0  ±  0	6.89  ±  1.29	0.66 ± 0.10
BDL and phlebotomy later	8	8.01  ±  3.1a	0.71  ±  0.16	5.97  ±  0.45	0.56 ± 0.07
Phlebotomy and BDL later	8	-3.29  ±  20.1	0.80  ±  0.15	7.02  ±  1.68	0.69 ±0.13

¹Mean phlebotomy volume for one phlebotomy session per rat;

^aP ≤ 0.01 vs BDL only.

Table 2 Liver function tests and enzymes in different groups of rats (mean±SE)

Subgroup	n	GGT(IU/L)	ALK- P(IU/L)	Bilirubin(µmol/L)	GPT(IU/L)	Albumin(G/L)
Sham only	5	7.0 ± 2.01	254 ± 321	4.6 ± 1.81	53 ± 112	28.0 ± 1.11
Sham and phlebotomy later	4	6.0 ± 1.4	305 ± 58	4.0 ± 0.8	66 ± 8	29.2 ± 2.2
Phlebotomy and sham later	6	14.3 ± 10.3	305 ± 61	41.7 ± 39.0	55 ± 20	28.4 ± 2.3
BDL only	9	74.0 ± 63.4	580 ± 174	106.0 ± 57.0	115 ± 50	24.3 ± 2.2
BDL and phlebotomy later	8	65.9 ± 17.9	578 ± 75	104.0 ± 16.2	125 ± 22	29.0 ± 2.01
Phlebotomy and BDL later	8	64.3 ± 17.6	665 ± 166	109.0 ± 39.0	144 ± 40	23.7 ± 2.1

¹P ≤ 0.007,

²P ≤ 0.02 vs BDL only GGT = Gamma-glutamyl transpeptidase, ALK-P = Alkaline phosphatase, GPT = Alanine aminotransferase. No difference in the levels of serum parameters like calcium, phosphor, urea, creatinine and electrolytes were found between the different groups (data not shown).

Table 3 Histological grading and staging and bile duct proliferation score in different groups of rats (mean±SE)

Subgroup	n	Periportal necrosis	Portal inflammation	Lobular necrosis	Fibrosis	Bile duct proliferation
Sham only	5	0.00 ± 0.00	0.40 ± 0.55	1.00 ± 1.22	0.00 ± 0.001	0.00 ± 0.001
Sham and phlebotomy later	4	0.00 ± 0.00	1.25 ± 1.26	0.25 ± 0.50	0.00 ± 0.00	0.00 ± 0.00
Phlebotomy and sham later	6	0.00 ± 0.00	1.00 ± 0.00	0.50 ± 0.55	0.67 ± 0.52	0.00 ± 0.00
BDL only	9	0.89 ± 1.27	1.33 ± 1.00	2.11 ± 1.83	2.89 ± 0.78	2.67 ± 0.50
BDL and phlebotomy later	8	0.13 ± 0.35	1.00 ± 0.00	3.00 ± 1.85	2.75 ± 0.71	2.88 ± 0.35
Phlebotomy and BDL later	8	2.17 ± 1.33	2.67 ± 0.82	4.00 ± 0.00	3.33 ± 0.52	2.83 ± 0.41

¹P ≤ 0.003 vs BDL only.

Table 4 Hepatic prostaglandin E2 (PGE2) generation, myeloperoxidase (MPO) activity and hydroxyproline (HP) content in different groups of rats (mean±SE)

Subgroup	n	PGE2 generation(ng/g liver)	HP content(ng/g liver)	MPO activity(units/g liver)
Sham only	5	14.8 ± 9.9	8.59 ± 0.551	0.45 ± 0.14
Sham and phlebotomy later	4	14.7 ± 5.2	8.11 ± 1.03	0.31 ± 0.12
Phlebotomy and sham later	6	6.9 ± 3.2	8.02 ± 1.00	0.18 ± 0.05
BDL only	9	8.5 ± 4.5	11.00 ± 0.97	0.59 ± 0.21
BDL and phlebotomy later	8	3.9 ± 2.11	9.80 ± 1.68	0.52 ± 0.10
Phlebotomy and BDL later	8	3.4 ± 2.31	11.90 ± 1.40	0.27 ± 0.171

¹P ≤ 0.018 vs BDL only.

Table 5 Hepatic iron content and concentration in different groups of rats (mean±SE)

Subgroup	n	Iron concentration(µcg/g/liver)	Iron content(µcg/total liver)
Sham only	5	91.8 ± 12.9	861.3 ± 158.9
Sham and phlebotomy later	4	40.1 ± 8.83	315.1 ± 63.33
Phlebotomy and sham later	6	55.0 ± 19.7	358.9 ± 104.4
BDL only	9	80.9 ± 17.2	1122.1 ± 358.6
BDL and phlebotomy later	8	37.4 ± 6.91	477.2 ± 85.11
Phlebotomy and BDL later	8	49.0  ± 29.6	628.0 ± 224.72

¹P ≤ 0.0001,

²P ≤ 0.012 vs BDL only;

³P  ≤ 0.016 vs sham only.

Table 6 Serum iron and transferrin in different groups of rats (mean±SE)

Subgroup	n	Serum iron (µg/dL)	Transferrin (mg/dL)
Sham only	5	163 ± 21	446 ± 46
Sham and phlebotomy later	4	144 ± 38	459 ± 17
Phlebotomy and sham later	6	160 ± 20	482 ± 103
BDL only	9	205 ± 71	640 ± 128
BDL and phlebotomy later	8	197 ± 57	732 ± 65
Phlebotomy and BDL later	8	215 ± 116	701 ± 83