The Expression And Effection Of Mast Cells And Cannabinoid CB2 Receptors In Interstitial Cystitis

Objective:To explore the the expression and effection of mast cells and cannabinoid CB2 receptors in interstitial cystitis through the detection of mast cells and cannabinoid CB2 receptors in interstitial cystitis (IC) model and the cannabinoid receptor agonist IC model expression, and seek new targets for the treatment of IC.Methods:1. Grouping:choose 30 SPF female SD rats, randomly divided into 3 groups: A.control group:10; B.IC group:10, C. IC intervention group:10.2. Methods:(1) infusion of 0.9% saline group C, Retention of 45 min, discharged after treatment with phosphoric acid buffer solution (PBS) 3 times, do not do special treatment for 1 consecutive weeks. (2) group B transurethral bladder perfusion 10 mg/ml of protamine sulfate 1 ml, keep 45 min, after row of all drugs in phosphate buffer solution (PBS) wash three times;Perfusion 750 ug/ml of lipopolysaccharide 1 ml, reserved for 30 min, do drugs after 3 times is washed with PBS, pull out the catheter, don’t do special handling, one week in a row. (3) the group A transurethral bladder perfusion 10 mg/ml of protamine sulfate 1 ml, keep 45 min, after row of all drugs in phosphate buffer solution (PBS) wash three times;Perfusion 750 ug/ml of lipopolysaccharide 1 ml, reserved for 30 min, do drugs after 3 times is washed with PBS, pull out the catheter,4h after intraperitoneal injection of 2 ml0.5 mg/ml; Cannabinoid receptor agonist CB2 agonist JWHO15, (5 mgjwhol5 dissolved in 10 ml20% of dimethyl sulfoxide), once a day for 1 week. One week immediately executed after three groups of rats, take the urinary bladder and the operation.(1) by HE staining observation research object IC pathology in animal model;(2) toluidine blue stain test of rat bladder MC number;(3) calculation cannabinoid CB2 immunohistochemical double staining positive cells and the relative expression quantity;(4) WESTERN-BLOT detection cannabinoid CB2 receptors.Results:1. In the blank group, the bladder mucosa and the mucosa were intact, and a small amount of inflammatory cell infiltration was found. In the IC group and the IC intervention group, the bladder mucosa damage was obvious, the mucosa layer appeared continuity damage, the lower layer of the mucous membrane, the bladder wall tissue showed a wide range of bleeding and congestion.2. MC by toluidine blue stained purple red, rats in the blank group bladder visible a small amount of MC; IC group, the bladder mucosa and submucosa shows a lot of MC expression (P<0.05, vs control group), after adding cannabinoid receptor agonist IC intervention rats bladder mucosa MC expression decreased, but still higher than that of blank group (P<0.05.vs in the blank group; P<0.05.vs cystitis group). Three groups of rat bladder muscle layer on MC less, and no significant difference (P>0.05).3. Immunohistochemical staining visible three grouds rats bladder mucosa have cannabinoid CB2 receptor expression; IC group, the bladder mucosa was significantly lower than that in control group (P<0.05, vs control group), after adding the cannabinoid receptor agonist, IC intervention group rats bladder mucosa of cannabinoid CB2 receptor was increased, but still below the blank group, higher than the IC group (P<0.05.vs blank group; P<0.05.vs the IC group).4. Western blot results, three groups of rat bladder mucosa were cannabinoid CB2 receptor expression; interstitial cystitis, bladder mucosal expression was significantly lower than control group (P<0.05, vs control group), after adding cannabinoid receptor agonist agent jwh015 intervention rats bladder mucosa marijuana (CB2 receptor expression increased, higher than interstitial bladder group, but still below the blank group (P<0.05.vs in the blank group; P<0.05.vs cystitis group). Three groups of rat muscle layer of cannabinoid CB2 receptor expression is less, and no significant difference (P>0.05).Conclusions:1. This experiment through the urethra and bladder instillation of protamine sulfate protein and lipopolysaccharide can successfully constructed rat interstitial cystitis (IC) animal model, consistent with the pathological features of human IC.2. In group IC, the MC of bladder tissue increased obviously, and the removal of MC was significantly increased, and there was a small amount of fibrosis in the muscle layer of the bladder. It was speculated that MC was closely related to the occurrence and development of IC3. In the IC intervention group, a large amount of MC was observed in bladder tissue, but the expression of CB2 receptor was lower, and the expression of receptor was higher. The CB2 cannabinoid receptor agonist agent can inhibit MC expression suggest that slow down and stop the IC progress, may become a new target for the treatment of IC.