Isoniazid-induced liver injury and immune response in mice

Figures & data

Table 1. Food consumption by mice.

Mouse strain	Food consumption (g/day)	Animal weight range (g)	Average NH/Pyr intake (mg/kg/day)
Male Balb/c
INH (0.1%)	3.1 ± 0.2	18–21	147–172
INH (0.15%)	3.6 ± 0.4	18–21	257–300
Female Balb/c
INH (0.1%)	3.5 ± 0.5	17–20	175–206
INH (0.15%)	2.7 ± 0.3	15–17	238–270
INH (0.2% + Pyr)	1.9 ± 0.4	13–17	224–292/56–73
Male Cbl-b^−/−
Control	4.0 ± 0.5	23–28
INH	3.8 ± 0.5	21–23	330–362
INH + Pyr	3.8 ± 0.4	21–23	330–362/83–90
Female Cbl-b^−/−
Control	4.5 ± 0.4	18–21
INH	3.8 ± 0.3*	16–18	422–475
Female PD1^−/−
Control	4.1 ± 0.4	15–25
INH (0.1–0.2%)	1.1–2.4*	14–20	110–170
Female C3H  + INH (0.2%)	4.5 ± 0.4	20–21	428–450
Female Rag^−/− + INH (0.2%)	1.9 ± 0.4	15–18	280–430

INH was given to mice at doses of 0.10, 0.15, or 0.20% [w/w] in food. Pyr = pyridoxine • HCl given to mice at 0.05% [w/w] in food. Values represent mean [±SE] of 4 mice/group except female C3H and female Rag^−/− mice (n = 3/group). Data were analyzed using a Mann-Whitney U test. * Significantly different from control group at p < 0.05.

Figure 1. GLDH activities, body weights, and INH covalent binding in INH-treated Balb/c mice. (a) GLDH in male mice treated at 0.10 or 0.15% INH [w/w] in food for 5 weeks. (b) GLDH in female mice treated at 0.10 or 0.15% of INH [w/w] in food for 3 weeks or treated with 0.2% of INH + 0.05% pyridoxine•HCl (Pyr) [w/w] in food for 2 weeks. (c, d) Body weights of treated mice. (e, f) Covalent binding of INH in livers of male and female mice. Values represent mean (±SE) of four mice/group. Analyzed for statistical significance by a Mann-Whitney U test; *p < 0.05.

Figure 2. Comparison of INH covalent binding in livers of different mice. (A) Male Cbl-b^−/− mice treated with INH (0.2% or 0.2% + 0.05% pyridoxine•HCl [w/w] in food, n = 3). (B) Female (n = 4) and male (n = 3) Cbl-b^−/− mice treated with 0.2% INH [w/w] in food. (C) Female C57BL/6 and Cbl-b^−/− mice treated with 0.2% INH [w/w] in food (n = 4). (D) Female Cbl-b^−/− control (n = 2), Cbl-b^−/− (n = 4), or C3H mice (n = 3) treated with 0.2% INH [w/w] in food.

Figure 3. GLDH activities and body weights in INH-treated Cbl-b^−/− mice. (A) GLDH in male mice treated with 0.2% INH [w/w] in food or 0.2% INH + 0.05% pyridoxine•HCl (INH + Pyr) [w/w] in food for 5 weeks. (B) GLDH in female mice treated with INH at 0.2% [w/w] in food for 5 weeks. (C, D) Body weights of male and female INH-treated mice. Values represent mean (±SE) from four mice/group. Analyzed for statistical significance by two-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001.

Figure 4. H&E and immunohistochemical staining for anti-CD45R, F4/80, and PCNA in livers of Cbl-b^−/− mice treated with INH (0.2% w/w in food) for 5 weeks. Control = untreated mice (n = 4); INH = mice treated with INH that did not develop abnormal liver histology (n = 3); INH steatosis = one mouse treated with INH that developed significant abnormal liver histology (n = 1). Red arrow = lymphocyte infiltration, yellow arrow = microvesicular steatosis, green arrow = macrovesicular steatosis, blue arrow = focal necrosis, orange arrow = cholestasis. 40× magnification for H&E; 20× magnification for CD45R, F4/80, and PCNA.

Figure 5. Serum concentrations of IL-1α and IL-12 (p70) in Cbl-b^−/− control or mice treated with INH for 5 weeks. Values represent mean (±SE) from four mice/group. INH was given at 0.2% INH [w/w] in food. Analyzed for statistical significance by a Mann-Whitney U test; *p < 0.05.

Figure 6. GLDH activity in female PD^−/− and Rag^−/− mice treated with INH. (A) INH was given at 0.2% [w/w] in food until Week 2 wherein the dose was decreased to 0.1% [w/w] in food for 1 week (until Week 3) because of weight loss. Mice were then put back on 0.2% INH [w/w] from Week 3–5. (B) Female C57BL/6 control or Rag^−/− mice were treated with INH at 0.2% [w/w] for up to 12 weeks. Values represent mean (±SE). Analyzed for statistical significance by two-way ANOVA. A p value <0.05 was considered significant (*p < 0.05; **p < 0.01; ***p < 0.001).