Experimental in vivo model to evaluate the impact of Cernitin™ on pain response on induced chronic bladder inflammation

Figures & data

Table 1. Experimental design.

Group	Induction of cystitis by injection of CYP (i.p.)	Treatments (p.o.)	Number of animals (n)
1	CYP	Vehicle 1 (H2O)	10
2	CYP	Vehicle 2 (DMSO)	10
3	CYP	T60 (22.5 mg/kg)	10
4	CYP	GBX (1.2375 mg/kg)	9
5	CYP	T60 + GBX (22.5 + 1.2375 mg/kg)	9
6	CYP	Gabapentin (100 mg/kg)	10

Test substances and concentrations are used in the study. CYP: cyclophosphamide; i.p.: intraperitonial; p.o.: per oral.

Figure 1. Experimental designs used in the study. Vehicles 1 and 2, gabapentin and test substances as indicated in Table 1. was administered on day (D) 7, D8, D9 and D10. Von Frey test was first performed at day 7 (before pharmacological treatment) to confirm presence of visceral pain and then at day 10 to analyze compounds effect on CYP-induced chronic bladder pain.

Figure 3. Mean expression level of COX-2 and MCP-1 in the bladder tissue homogenates. (A) a significant decrease in COX-2 levels was observed in the Cernitin™T60 (**p < .01) and Cernitin™ GBX (*p < .05) groups compared to vehicle-treated groups, combination treatment did not enhance the effect of Cernitin™ (*p < .05). (B) MCP-1 levels remained unchanged. Gabapentin (Gaba)-treated group produced a significant reduction of COX-2 level (*p < .05). Data are presented as mean ± SD (n = 5).

Table 2. Hematoxylin–eosin staining analyses of tissue sections for presence of bladder inflammation and edema performed in five animals from each treatment group.

	Edema	Inflammation
H2O	5 (2)	5 (2)
DMSO	4 (2)	4 (2)
T60	1 (1)	1 (1)
GBX	1 (1)	2 (1)
T60 + GBX	1 (1)	2 (1)
Gabapentin	3 (1)	1 (1)

Numbers in the column indicate the number of rats presented with the conditions and the number in the parentheses indicating the scores.

Figure 4. Immunohistochemical analysis of the bladder tissues. Effects of Cernitin™ on the expression of (A) CD45, (B) PGD2 and (C) SP. T + G = T60 + GBX. Immunostaining was performed in the five animals mentioned in Table 2. intensity was determined by color intensity as follows; strongly stained as score 3, moderately stained as score 2 and weakly stained as score 1, no staining as score 0.