Boric acid is associated with the suppression of apoptosis and endoplasmic reticulum stress in rat model of paracetamol -induced hepatotoxicity

Figures & data

Table

sCasp1-80F	TTTTATTGCTTTCTGCTCTTCAAC
sCasp1-80R	TGATGAGTGACTCAATGAAGAGAGAT
sCasp9-80F	TTGTGTCCTACTCCACCTTCC
sCasp9-80R	GATGTACCAGGAACCGCTCT
sCasp3-80F	CCGACTTCCTGTATGCTTACTCTA
sCasp3-80R	CATGACCCGTCCCTTGAA
sATF6-80F	GACAAATAGCCAGCAGAAAACC
sATF6-80R	TGAGAATTCGAGCCCTGTTC
sRELA-26F	GCCATGGACGATCTGTTTC
sRELA-26R	GCTGCTCAATGATCTCCACA
sCasp8-26F	ATATCAGTCGGCGGGATCT
sCasp8-26R	TTCTGAGAGCTTAAAGAGCATAACC
sNFKB-26F	GCCATGGACGATCTGTTTC
sNFKB-26R	GCTGCTCAATGATCTCCACA
ERN1-18F	CCAAACATCGAGAAAACGTG
ERN1-18R	TCTGAAGTCTGGCCAACTAGG
sINOS-18-F	AGGTTGGAGGCCTTGTGTC
sINOS-18-R	GCTTCAGAATGGGGAGCTG
sBCL2-2F	GTACCTGAACCGGCATCTG
sBCL2-2R	CTGAGCAGCGTCTTCAGAGA
sBCL2-XL-2F	TGACCACCTAGAGCCTTGGA
sBCL2-XL-2R	TTCCCGTAGAGATCCACAAAA
sPERK-6F	CGAAGGCCTTGGAGTCTGTA
sPERK-6R	AAGGGCTTCCATTTGATCGT
sCasp12-94F	TGTGTTGAGCCATAGATGGAA
sCasp12-94R	TTGCCTACCTTCCAAGAAATG

Table 1. The effects of the boric acid and paracetamol on Aspartate transaminase (AST) and Alanine Aminotransferase (ALT) levels in serum of rats.

	AST (u/l)	ALT (u/l)
Control	99.3 ± 6.46^a	24.2 ± 2.65^a
Paracetamol	160.7 ± 17.12^b	68.5 ± 3.83^b
Paracetamol + 50 boron	138.4 ± 19.42^c,d	36.5 ± 6.22^c,d
Paracetamol + 100 boron	104.6 ± 13.64^d	31.2 ± 3.39^a,c,d
Paracetamol + 200 boron	107.5 ± 16.63^d	35.2 ± 7.3^c,d
Paracetamol + NAC	100.3 ± 9.14^a	23 ± 2.01^a
200 boron	103.7 ± 9.95^a,d	27.6 ± 2.75^a,d

Different letters in the same column are statistically significant from each other. P < 0.05.

Serum AST, ALT levels different letters in the same column are statistically significant from each other.

Table 2. The effects of the boric acid and paracetamol on Tumour Necrosis Factor (TNF –α), interleukin 1β (IL-1β), interleukin 18 (IL-18) levels in liver of rats.

	TNF –α (ng/l)	IL-1β (ng/l)	IL-18 (pg/l)
Control	355.10 ± 16.27^a	2.86 ± 0.52^a	34.09 ± 1.6^a
Paracetamol	671.92 ± 18.11^d	2.96 ± 0.01^a	82.93 ± 3.21^f
Paracetamol + 50 boric acid	453.24 ± 24.16^b,c	2.70 ± 0.32^a,b	66.47 ± 4.62^e
Paracetamol + 100 boric acid	445.98 ± 24.58^b,c	2.81 ± 0.33^a,b	54.27 ± 11.34^d
Paracetamol + 200 boric acid	494.80 ± 27.58^c	2.46 ± 0.19^b,c	50.98 ± 6.45^c,d
Paracetamol + NAC	417.27 ± 29.18^a,b	2.29 ± 0.23^c	44.75 ± 8.66^b,c
200 boric acid	375.85 ± 16.36^a	2.68 ± 0.34^a,b	40.99 ± 3.31^a,b

Different letters in the same column were statistically significant. P < 0.05.

Liver tissue cytokine levels. Different letters in the same column were statistically significant. P < 0,05 was considered significant.

Table 3. The effects of the boric acid and paracetamol on Tumour Necrosis Factor (tnf –α), interleukin 1β (ıl-1β), interleukin 18 (ıl-18) levels in serum of rats.

	TNF –α (ng/l)	IL-1β (ng/l)	IL-18 (pg/l)
Control	115.9 ± 6.1^a	0.78 ± 0.12^a	15.7 ± 2.1^a
Paracetamol	370.9 ± 8.22^d	0.86 ± 0.11^a	7.1 ± 4.2^b
Paracetamol + 50 boric acid	210.8 ± 7.26^b,c	0.76 ± 0.12^a,b	10.47 ± 3.9^c
Paracetamol + 100 boric acid	220.9 ± 9.24^b,c	0.71 ± 0.13^a,b	11.27 ± 2.8^c
Paracetamol + 200 boric acid	294.8 ± 7.9^c	0.66 ± 0.17^b,c	12.98 ± 4.5^c
Paracetamol + NAC	247.27 ± 9.8^a,b	0.69 ± 0.18^c	14.15 ± 5.1^a,c
200 boric acid	129.9 ± 6.7^a	0.81 ± 0.14^a,b	13.19 ± 3.11^a,c

Different letters in the same column were statistically significant. P < 0.05.

Serum cytokine levels. Different letters in the same column were statistically significant. P < 0,05 was considered significant.

Table 4. The effects of the boric acid and paracetamol on Liver Tissue Malondialdehyde (MDA), Glutathione (GSH), Glutathione peroxidase (GPX), Nitric Oxide (NO), superoxide dismutase (SOD), catalase (CAT) and 8-Hydroxy-2'-deoxyguanosine (8-OHdG) levels in rats.

	MDA nmol/g	NO µmol/l	SOD ng/ml	CAT ng/ml	8-OHdG ng/ml	GSH nmol/ml	GP xng/ml
Control	424.61 ± 9.24^a	2.97 ± 0.91^a	360.20 ± 19.1^a	4.67 ± 0.29a	24.20 ± 2.65a	11.35 ± 0.92^a	360.20 ± 20.81^a
Paracetamol	721.27 ± 16.31^d	5.68 ± 0.82^b	101.60 ± 4.69b	2.41 ± 0.24c	68.50 ± 3.83b	4.88 ± 0.51^d	101.60 ± 4.69^b
Paracetamol + 50 boric acid	530.25 ± 14.96^b,c	3.67 ± 0.67^b	293.90 ± 6.79^c	3.70 ± 0.17b	36.50 ± 6.22^c,d	9.87 ± 0.79^a	193.90 ± 6.79^c
Paracetamol + 100 boric acid	535.98 ± 14.98^b,c	3.46 ± 0.45^c	255.80 ± 8.66^c	4.44 ± 0.30a	31.20 ± 3.39^a,c,d	11.20 ± 1.11^a,b,c	255.80 ± 18.66^d
Paracetamol + 200 boric acid	524.20 ± 17.48^c	3.64 ± 0.20^b	246.00 ± 5.25^c	4.41 ± 0.21a	35.20 ± 7.30^c,d	10.77 ± 0.91^a,b,c	346.00 ± 25.25^a
Paracetamol + NAC	527.17 ± 19.18^a,b	2.54 ± 0.14^b	210.90 ± 8.99^c	4.52 ± 0.16a	23.00 ± 2.01^a	11.45 ± 0.79^c	210.90 ± 8.99^c
200 boric acid	475.35 ± 17.26^a	2.59 ± 0.53^a	236.40 ± 8.32^,c	4.62 ± 0.48a	24.60 ± 2.75^a	11.86 ± 1.23^b	336.40 ± 28.32^a

Different letters in the same column are statisticall significant. p < 0.05.

Liver tissue MDA, GSH, GPx, NO, SOD, CAT and 8-OHG different letters in the same column were statistically significant from each other. P < 0,05 was considered significant.

Table 5. The effects of the boric acid and paracetamol on serum Malondialdehyde (MDA), Glutathione (GSH), Glutathione peroxidase (GPX), Nitric Oxide (NO), superoxide dismutase (SOD), catalase (CAT) and 8-Hydroxy-2'-deoxyguanosine (8-OHdG) levels in rats.

–	MDA nmol/g	NO µmol/l	SOD ng/ml	CAT ng/ml	8- OHdG ng/ml	GSH nmol/mı	GP xng/ml
Control	0.21 ± 0.85^a	3.15 ± 0.73^a	79.21 ± 0.62^a	3.15 ± 0.73^a	33.98 ± 8.049^a	2.19 ± 3.35^a	328.95 ± 31.16^a
Paracetamol	0.53 ± 0.13^b	10.49 ± 0.37^b	58.43 ± 0.54^c	10.89 ± 0.67^b	71.77 ± 3.65^c	0.03 ± 0.3^b	440.63 ± 14.17^b
Paracetamol + 50 boric acid	0.41 ± 0.65^c	5.72 ± 0.16^c	78.16 ± 0.42^a	5.71 ± 0.16^c	30.55 ± 3.87^a,b	1.32 ± 2.02^c	413.46 ± 23.28^b
Paracetamol + 100 boric acid	0.42 ± 0.65^c	6.45 ± 0.80^d	67.56 ± 1.36^b	5.45 ± 0.80^d	30.47 ± 3.97^a,b	1.01 ± 1.36^c	345.75 ± 21.34^a,c
Paracetamol + 200 boric acid	0.56 ± 0.17^b	3.14 ± 0.08^a	78.08 ± 0.87^a	3.20 ± 0.88^a	28.62 ± 1.83^b	2.30 ± 2.69^a	319.21 ± 25.41^c
Paracetamol + NAC	0.24 ± 0.72^b	2.74 ± 0.56^a	78.71 ± 1.07^a	2.94 ± 0.96^a	27.21 ± 7.75^b	2.11 ± 1.52^a	279.78 ± 32.89^a
200 boric acid	0.22 ± 0.82^a	2.99 ± 0.48^a	77.41 ± 2.01^a	2.98 ± 0.48^a	29.91 ± 6.55^a,b	2.14 ± 1.41^a	319.21 ± 15.31^a,c

Different letters in the same column are statisticall significant. p < 0.05.

Serum MDA, GSH, NO, 8-OHG and erythrocyte, SOD, CAT and GPx levels. Different letters in the same column are statistically significant. P < 0,05 is considered significant.

Table 6. The effects of the boric acid and paracetamol on mRNA expression of activating transcription factor 6 (ATF-6), B-cell lymphoma 2 (BCL2), B-cell lymphoma-extra (Bcl-x), Caspase 1 (CASP1), Caspase 12(CASP12), Caspase 3 (CASP3), Nuclear Factor kappa B (NF-HB), cyclooxygenase-2 (COX-2).

	ATF-6	BCL2	BCLX	CASP1	CASP12	CASP3	ASP8	NF-KB	COX-2
Paracetamol	5.01 ± 0.62^a	0.94 ± 0.08^f	0.59 ± 0.17^f	4.48 ± 0.63^a	6.77 ± 0.63^a	4.42 ± 0.62^a	3.68 ± 0.41^a	6.78 ± 0.22^a	3.28 ± 0.13^a
Paracetamol + 50 boric acid	4.24 ± 0.54^b	1.24 ± 0.12^e	1.02 ± 0.20^e	2.85 ± 0.73^b	5.11 ± 0.51^b	2.88 ± 0.38^b	1.82 ± 0.26^b	4.32 ± 0.31^b	1.45 ± 0.43^b
Paracetamol + 100 boric acid	2.91 ± 0.49^c	1.80 ± 0.16^d	1.51 ± 0.11^d	1.51 ± 0.24^c	2.50 ± 0.47^c	2.12 ± 0.18^c	1.26 ± 0.15^c	4.31 ± 0.25^c	1.31 ± 0,44^c
Paracetamol + 200 boric acid	1.83 ± 0.26^d	2.40 ± 0.24^c	2.09 ± 0.15^c	0.94 ± 0.16^d	1.21 ± 0.24^d	1.53 ± 0.19^d	1.03 ± 0.06^d	4.02 ± 0.10^d	0.89 ± 0.12^d
Paracetamol + NAC	1.27 ± 0.12^e	3.44 ± 0.36^b	3.46 ± 0.21^b	0.54 ± 0.13^de	0.71 ± 0.17^e	1.15 ± 0.11^e	0.77 ± 0.09^e	1.49 ± 0.08^e	0.69 ± 0.15^de
200 boric acid	0.66 ± 0.18^f	4.12 ± 0.51^a	4.91 ± 0.49^a	0.22 ± 0.03^d	0.27 ± 0.07^f	0.60 ± 0.27^f	0.39 ± 0.08^f	0.69 ± 0.09^f	0.34 ± 0.02^d

Different letters in the same column are statistically significant from each other P < 0.05.

Results of liver tissue molecular analysis, different letters in the same column are statistically significant from each other. P < 0,05 was considered significant.

Table 7. The effects of the boric acid and paracetamol on mRNA expression of Caspase 9 (CASP9), endoplasmic reticulum to nucleus signalling 1 (ERN1), Inducible nitric oxide synthase (iNOS), endoplasmic reticulum kinase (PERK).

	CASP9	ERN1	INOS	PERK	RELA-1
Paracetamol	4.10 ± 0.50^a	5.64 ± 0.53^a	5.53 ± 0.69^a	4.30 ± 0.70^a	3.82 ± 0.37^a
Paracetamol + 50 boric acid	2.22 ± 0.14^b	4.02 ± 0.61^b	4.00 ± 0.77^b	3.51 ± 0.64^b	2.65 ± 0.44^b
Paracetamol + 100 boric acid	1.65 ± 0.23^c	2.41 ± 0.39^c	2.31 ± 0.86^c	1.89 ± 0.44^c	1.79 ± 0.11^c
Paracetamol + 200 boric acid	1.20 ± 0.08^d	1.63 ± 0.18^d	0.97 ± 0.34^d	1.14 ± 0.10^d	1.41 ± 0.15^d
Paracetamol + NAC	0.99 ± 0.06^d	1.11 ± 0.14^e	0.38 ± 0.08^e	0.72 ± 0.23^d	1.00 ± 0.13^e
200 boric acid	0.66 ± 0.16^e	0.65 ± 0.19^f	0.16 ± 0.03^e	0.25 ± 0.03^e	0.61 ± 0.11^f

Different letters in the same column are statistically significant from each other P < 0.05.

Results of molecular analysis of liver tissue, different letters in the same column were statistically significant from each other. P < 0,05 was considered significant.

Figure 1. The effects of the boric acid and paracetamol on histopathological alterations in rat liver tissue. Arrow: necrosis and mononuclear cell infiltrations in the pericentral region of the liver, arrowhead: sinusoidal dilatation and hyperemia, A: Control; B: Paracetamol; C: Paracetamol + 50 boric acid; D: Paracetamol + 100 boric acid; E: Paracetamol + 200 boric acid; F: Paracetamol + NAC; G: 200 boric acid. All figures are stained with H&E. 20x, and 100 µm were used as the original magnifications. H: Damage score of histopathological alterations. 0-no damage, 1-slight, 2-moderate, 3-severe, or 4-most damaged. MNI: mononuclear cell infiltrations. ^a,b,c,d:Different letters in the same column were statistically significant, p < 0.05.