Sub-chronic toxicological studies of transition metal complexes of naproxen on sprague-dawley rats

Peer review under responsibility of Alexandria University Faculty of Medicine.

Available online 18 January 2017

Figures & data

Table 1 Experimental design of naproxen metal complexes.

Groups	Test samples	No of animals
Group I	Control	8
Group II	Naproxen	8
Group III	Naproxen copper complex	8
Group IV	Naproxen cobalt complex	8
Group V	Naproxen iron complex	8
Group VI	Naproxen silver complex	8
Group VII	Naproxen zinc complex	8

Table 2 The number of ulcers was counted. Ulcer scoring was undertaken21 as following manner.

Ulcer score	Descriptive/observation
0	No ulcer/normal colored stomach
0.5	Red coloration
1.0	Superficial (spot) ulcer
1.5	Hemorrhagic streak
2.0	Deep ulcer
3.0	Perforation

Fig. 1 Microscopic evaluation of ulceration of (a) control, (b) naproxen, (c) naproxen copper complex, (d) naproxen cobalt complex, (e) naproxen iron complex, (f) naproxen silver complex, (g) naproxen zinc complex of the rat groups.

Table 3 Hematological parameter changes of male rats during treatment of test compounds.

Parameters	Control	Standard (Naproxen)	Nap-Cu	Nap-Co	Nap-Fe	Nap-Ag	Nap-Zn
WBC (10^9/L)	4.9 ± 0.36	7.44 ± 0.31^*	4.9 ± 0.84	5.44 ± 0.61	6.72 ± 0.81	5.6 ± 0.49	5.97 ± 0.57
RBC (10^12/L)	5.66 ± 0.17	4.72 ± 0.32^*	5.98 ± 0.23	6.71 ± 0.22	5.88 ± 0.44	5.51 ± 0.21	5.67 ± 0.46
Platelets (10^11/L)	5.88 ± 0.11	6.79 ± 0.61	5.01 ± 0.92	5.11 ± 0.11	5.32 ± 0.20	4.97 ± 0.66	5.1 ± 0.26
Neutrophils (10^9/L)	0.84 ± 0.07	1.57 ± 0.23^***	0.95 ± 0.08	1.22 ± 0.33	1.11 ± 0.02	0.98 ± 0.07	1.04 ± 0.06
Lymphocytes (10^9/L)	3.92 ± 0.76	5.66 ± 0.27^*	4.78 ± 0.22	4.91 ± 0.44	4.88 ± 0.18	4.55 ± 0.12	4.78 ± 0.39
Monocytes (10^9/L)	0.05 ± 0.007	0.14 ± 0.02^*	0.1 ± 0.02	0.1 ± 0.02	0.12 ± 0.01	0.07 ± 0.02	0.07 ± 0.02
Hemoglobin (gm/dL)	11.71 ± 0.71	9.11 ± 0.80^**	11.01 ± 0.34	11.19 ± 0.20	10.61 ± 0.33	10.64 ± 0.17	10.88 ± 0.22
Hematocrit (HCT) (%)	32.62 ± 1.4	25.96 ± 1.2^**	32.88 ± 0.51	30.91 ± 0.58	29.72 ± 1.4	30.89 ± 0.51	31.03 ± 0.96
MCV (fL)	54.92 ± 1.7	54.64 ± 0.65	53.07 ± 0.71	53.01 ± 0.70	53.98 ± 1.4	53.94 ± 0.91	52.65 ± 0.71
MCH (pg)	19.98 ± 0.63	19.76 ± 0.31	18.01 ± 0.67	18.11 ± 0.06	19.13 ± 0.45	19.6 ± 0.34	18.2 ± 0.35
MCHC (gm/dL)	34.44 ± 0.31	33.96 ± 0.12	34.02 ± 0.09	34.63 ± 0.11	34.15 ± 0.23	34.01 ± 0.11	34.38 ± 0.19
RDW (%)	18.33 ± 2.7	18.44 ± 0.28	14.92 ± 0.48	15.07 ± 0.61	16.67 ± 1.4	16.12 ± 0.56	16.01 ± 0.31
MPV (fL)	4.11 ± 0.07	4.19 ± 0.07	4.11 ± 0.09	4.21 ± 0.11	4.19 ± 0.04	4.11 ± 0.07	4.31 ± 0.08
PDW (10 GSD)	16.27 ± 0.11	16.22 ± 0.08	16.33 ± 0.16	16.84 ± 0.27	16.21 ± 0.11	16.5 ± 0.17	16.51 ± 0.19
ESR (mm in 1st hour)	2.47 ± 0.27	2.34 ± 0.21	2.11 ± 0.11	2.21 ± 0.12	2.12 ± 0.11	2.1 ± 0.01	2.67 ± 0.21
Bleeding time (sec)	47.11 ± 3.7	44.11 ± 3.1	41.11 ± 2.1	47.11 ± 3.7	41.34 ± 3.1	41.47 ± 3.1	39.68 ± 3.7
Clotting time (min)	3.68 ± 0.07	3.74 ± 0.11	4.78 ± 0.07	4.1 ± 0.10	3.77 ± 0.06	4.92 ± 0.07	4.11 ± 0.08

Data were expressed as mean ± SEM. n = 8.

* p < 0.05 compared to control.

** p < 0.01 compared to control.

*** p < 0.001 compared to control.

Table 4 Hematological parameter changes of female rats during treatment of test compounds.

Parameters	Control	Standard (Naproxen)	Cu-Nap	Co-Nap	Fe-Nap	Ag-Nap	Zn-Nap
WBC (10^9/L)	4.8 ± 0.28	7.09 ± 0.43^*	5.7 ± 0.37	6.21 ± 0.71	6.17 ± 0.68	5.3 ± 0.63	5.93 ± 0.47
RBC (10^12/L)	5.82 ± 0.37	4.81 ± 0.28^*	6.32 ± 0.16	6.69 ± 0.13	5.75 ± 0.36	5.66 ± 0.13	5.9 ± 0.18
Platelets (10^11/L)	5.91 ± 0.30	6.59 ± 0.27	4.92 ± 0.25	5.04 ± 0.37	5.44 ± 0.25	5.02 ± 0.18	5.08 ± 0.22
Neutrophils (10^9/L)	0.87 ± 0.06	1.54 ± 0.21^***	0.98 ± 0.09	1.26 ± 0.12	1.14 ± 0.09	0.97 ± 0.09	1.02 ± 0.07
Lymphocytes (10^9/L)	3.87 ± 0.26	5.56 ± 0.29^*	4.63 ± 0.35	4.85 ± 0.59	4.93 ± 0.57	4.25 ± 0.42	4.83 ± 0.42
Monocytes (10^9/L)	0.06 ± 0.008	0.13 ± 0.02^*	0.1 ± 0.01	0.1 ± 0.01	0.11 ± 0.01	0.08 ± 0.01	0.08 ± 0.01
Hemoglobin (gm/dL)	10.98 ± 0.53	9.17 ± 0.44^**	11.34 ± 0.21	12.09 ± 0.23	10.77 ± 0.42	10.46 ± 0.18	10.67 ± 0.29
Hematocrit (HCT) (%)	32.16 ± 1.6	26.63 ± 1.4^**	33.41 ± 0.59	35.13 ± 0.58	31.27 ± 1.4	30.81 ± 0.51	31.03 ± 0.96
MCV (fL)	55.66 ± 1.5	55.54 ± 0.65	52.97 ± 0.51	52.59 ± 0.40	54.83 ± 1.2	54.49 ± 0.47	52.56 ± 0.40
MCH (pg)	19.06 ± 0.59	19.17 ± 0.26	17.99 ± 0.18	18.06 ± 0.13	18.93 ± 0.53	18.5 ± 0.14	18.1 ± 0.15
MCHC (gm/dL)	34.21 ± 0.25	34.53 ± 0.27	33.99 ± 0.11	34.36 ± 0.13	34.51 ± 0.23	33.93 ± 0.11	34.43 ± 0.13
RDW (%)	18.64 ± 2.0	18.84 ± 0.68	14.84 ± 0.57	15.07 ± 0.59	16.91 ± 1.3	15.34 ± 0.65	15.09 ± 0.41
MPV (fL)	4.19 ± 0.09	4.24 ± 0.09	4.16 ± 0.08	4.36 ± 0.13	4.11 ± 0.06	4.17 ± 0.08	4.24 ± 0.09
PDW (10 GSD)	16.36 ± 0.15	16.24 ± 0.09	16.41 ± 0.15	16.91 ± 0.32	16.44 ± 0.17	16.6 ± 0.16	16.67 ± 0.21
ESR (mm in 1st hour)	2.57 ± 0.29	2.43 ± 0.29	2.14 ± 0.14	2.14 ± 0.14	2.14 ± 0.14	2.0 ± 0.0	2.71 ± 0.28
Bleeding time (sec)	47.14 ± 3.9	47.14 ± 3.9	47.14 ± 2.1	47.14 ± 3.9	36.43 ± 3.0	38.57 ± 3.0	42.86 ± 3.9
Clotting time (min)	3.86 ± 0.09	3.86 ± 0.14	3.82 ± 0.08	4.0 ± 0.10	3.82 ± 0.07	3.96 ± 0.08	4.14 ± 0.09

Data were expressed as mean ± SEM. n = 8.

* p < 0.05 compared to control.

** p < 0.01 compared to control.

*** p < 0.001 compared to control.

Table 5 Biochemical parameter changes of male rats during treatment of test compounds.

Parameters	Control	Naproxen	Nap-Cu	Nap-Co	Nap-Fe	Nap-Ag	Nap-Zn
Glucose (mg/dl)	123.3 ± 3.4	125.6 ± 4.3	123.5 ± 4.1	123.2 ± 4.1	124.3 ± 3.8	123.9 ± 3.9	123.8 ± 4.2
Creatinine (mg/dl)	0.85 ± 0.05	0.9 ± 0.06	0.79 ± 0.08	0.94 ± 0.08	0.78 ± 0.03	0.81 ± 0.08	0.79 ± 0.05
Protein (g/dl)	7.32 ± 0.15	7.27 ± 0.18	7.48 ± 0.11	6.77 ± 0.21	7.39 ± 0.19	7.18 ± 0.08	7.51 ± 0.21
Albumin (g/dl)	4.28 ± 0.17	4.29 ± 0.45	4.13 ± 0.16	3.5 ± 0.06	4.12 ± 0.16	4.24 ± 0.12	4.26 ± 0.15
Total cholesterol (mg/dl)	66.7 ± 4.6	68.4 ± 5.2	65.6 ± 4.3	67.7 ± 4.9	66.2 ± 4.8	65.4 ± 5.6	67.1 ± 5.8
Triglycerides (mg/dl)	86.2 ± 3.6	84.7 ± 4.5	87.1 ± 4.1	85.9 ± 3.9	87.6 ± 5.3	85.6 ± 4.9	86.6 ± 5.5
Total bilirubin (mg/dl)	0.06 ± 0.3	0.05 ± 0.3	0.07 ± 0.2	0.05 ± 0.3	0.06 ± 0.4	0.06 ± 0.7	0.05 ± 0.4
Urea (mg/dl)	14.56 ± 0.25	15.07 ± 0.55	14.61 ± 0.63	14.47 ± 0.47	15.10 ± 0.76	14.45 ± 0.65	15.01 ± 0.45
Uric acid (mg/dl)	4.19 ± 0.43	4.27 ± 0.38	3.96 ± 0.51	4.31 ± 0.44	4.11 ± 0.51	4.38 ± 0.42	4.31 ± 0.40
ALT (IU/L)	37.4 ± 3.3	38.4 ± 3.3	36.8 ± 2.7	37.8 ± 5.3	37.2 ± 3.9	36.9 ± 3.5	38.2 ± 4.1
AST (IU/L)	185.3 ± 9.8	187.3 ± 10.8	182.2 ± 10.7	187.4 ± 12.6	184.7 ± 11.4	186.3 ± 13.6	183.6 ± 12.4
Alk phos (IU/L)	52.2 ± 3.2	54.1 ± 3.5	51.5 ± 2.8	52.5 ± 3.1	52.6 ± 3.5	53.4 ± 3.4	52.8 ± 3.3

Data were expressed as mean ± SEM. n = 8.

Table 6 Biochemical parameter changes of female rats during treatment of test compounds.

Parameters	Control	Naproxen	Nap-Cu	Nap-Co	Nap-Fe	Nap-Ag	Nap-Zn
Glucose (mg/dl)	121.1 ± 3.4	123.4 ± 3.2	124.1 ± 3.1	119.2 ± 4.1	122.1 ± 3.4	120.7 ± 4.4	124.8 ± 5.5
Creatinine (mg/dl)	0.83 ± 0.04	0.7 ± 0.05	0.77 ± 0.06	0.74 ± 0.04	0.78 ± 0.03	0.77 ± 0.08	0.82 ± 0.05
Protein (g/dl)	6.51 ± 0.21	6.27 ± 0.21	6.48 ± 0.31	6.27 ± 0.27	6.39 ± 0.12	6.58 ± 0.1	6.51 ± 0.29
Albumin (g/dl)	3.82 ± 0.25	3.92 ± 0.53	3.86 ± 0.34	3.72 ± 0.16	3.78 ± 0.23	3.84 ± 0.17	3.62 ± 0.19
Total cholesterol (mg/dl)	64.7 ± 3.6	66.4 ± 3.8	63.7 ± 2.9	65.8 ± 3.4	64.3 ± 3.6	63.9 ± 2.7	65.2 ± 3.2
Triglycerides (mg/dl)	84.4 ± 2.6	82.8 ± 3.2	85.3 ± 3.4	82.2 ± 4.1	85.5 ± 3.7	83.7 ± 3.4	81.9 ± 4.3
Total bilirubin (mg/dl)	0.05 ± 0.2	0.04 ± 0.4	0.06 ± 0.4	0.04 ± 0.3	0.05 ± 0.5	0.05 ± 0.3	0.04 ± 0.3
Urea (mg/dl)	12.56 ± 0.25	13.07 ± 0.43	12.61 ± 0.51	12.47 ± 0.42	13.10 ± 0.45	12.45 ± 0.31	13.01 ± 0.32
Uric acid (mg/dl)	3.49 ± 0.31	3.27 ± 0.47	3.56 ± 0.58	3.61 ± 0.52	4.0 ± 0.78	3.71 ± 0.61	3.93 ± 0.67
ALT (IU/L)	35.4 ± 2.9	36.4 ± 4.1	34.8 ± 3.7	35.8 ± 3.8	35.2 ± 4.9	34.9 ± 2.6	36.2 ± 3.9
AST (IU/L)	182.3 ± 10.3	184.3 ± 10.1	180.2 ± 9.7	185.4 ± 11.1	181.7 ± 9.9	183.3 ± 10.2	181.6 ± 11.1
Alk phos (IU/L)	50.5 ± 2.1	52.3 ± 3.4	49.8 ± 2.3	50.3 ± 3.7	50.6 ± 2.9	51.8 ± 2.2	51.9 ± 3.8

Data were expressed as mean ± SEM. n = 8.

Table 7 GI effects of different groups of rats under the experimental period.

Group	Observed parameters for male rat
	Number of ulcers (Mean ± SEM)	Average Ulcer score (Mean ± SEM)	Ulcer index (Mean ± SEM)	Inhibition of ulceration (%)
Control	–	–	–	–
Naproxen	2.1 ± 0.26	1.3 ± 0.09	13.4 ± 0.29	–
N-Cu	0.9 ± 0.14^***	0.8 ± 0.18	10.2 ± 0.30^***	23.88
N-Co	1.1 ± 0.14^*	0.8 ± 0.10	11.9 ± 0.20^*	11.19
N-Fe	1.1 ± 0.14^*	0.8 ± 0.10	11.9 ± 0.20^*	11.19
N-Ag	1.0 ± 0.31^**	1.0 ± 0.31	9.1 ± 0.62^***	32.09
N-Zn	1.1 ± 0.26^*	0.9 ± 0.25	10.6 ± 0.51^***	20.89
For female rat
Control	–	–	–	–
Naproxen	1.9 ± 0.21	1.1 ± 0.08	12.3 ± 0.27	_
N-Cu	0.8 ± 0.15^***	0.7 ± 0.17	10.8 ± 0.31^***	22.71
N-Co	1.3 ± 0.13^*	0.9 ± 0.10	12.1 ± 0.19^*	11.94
N-Fe	1.2 ± 0.13^*	0.9 ± 0.14	12.4 ± 0.48^*	12.79
N-Ag	1.4 ± 0.14^*	1.1 ± 0.10	12.3 ± 0.20^*	12.01
N-Zn	0.9 ± 0.34^***	1.3 ± 0.41	9.8 ± 0.72^***	32.45

Data were expressed as mean ± SEM. n = 8.

* p < 0.05 compared to standard.

** p < 0.01 compared to standard.

*** p < 0.001 compared to standard.