Study On The Role Of Mast Cell In Athongenesis Of Interstitial Cystitis

Objective1. To discuss whether there is a application value in the dagnosis of interstitial cystitis with the mast cells’ property and the level of PUF scale.in urinary bladder of IC patients.2. To investigate the role of estrogen and the mast cells in the pathogenesis of the interstitial cystitis patients, specifically with respect to activated bladder mast cells.3. To generate transgenic URO-OVA mice that express OVA as a "self" Ag on the bladder epithelium. In addition, we also generated URO-OVA/KitW-SH mice which express URO-OVA and mast cells are deficient also.URO-OVA and URO-OVA/KitW"SH mice develop bladder inflammation upon the introduction of OVA-specific OT-IT cells. To investigate the difference of Mast Cell in two groups. To discuss the role of the mast cells in the pathogenesis of IC.Materials and Methods1. Pieces were stained with0.5%toluidine blue and immobilizated with Osmic Acid. After observated and counted mast cells with light microscope and transmission electron microscope. We made a comparison on the PUF scale and the number and characteristics of mast cells between the two groups and also did statistical analysis upon the correlation between the PUF scale and the mast cell infiltration number.2. Pieces were analyzed blindly for the numbers of mast cells and the presence of estrogen receptors(ER). Immunohistochemical methods were used to detect ER and MC. And double immunohistochemical staining was used to manifest the expression of ER in interstitial cystitis MC.3. Transgenic URO-OVA mice and URO-OVA/KitW-SH mice are constructed and backcrossed with B6mice for six generations. The T cells from OT-I mice were injected i.v. into both transgenic URO-OVA mice and URO-OVA/KitW-SH group.Mice were sacrificed and the bladders were collected for inflammation analysis and mast cells analysis.Results 1. The mast cell’s number of the interstitial cystitis samples (28-76pieces/mm2) was significantly higher than that of the average persons’bladder tissues (7-15pieces/mm2)(Z=3.927, P<0.01);75.3%mast cells were in a state of being activated degranulation; the PUF scale of interstitial cystitis patients (17-35scores) was significantly higher than that of the healthy persons’(0-8scores)(t=14.736, P<0.01); the PUF scale of the patient group does not have a linear relation with the mast cell’s infiltrated number among the specimens (rs=-0.618. P=0.601).2. Bladder MC alone were shown to express high affinity ER in IC(3.77±3.03/HP) comparing with controls (0.88±0.76/HP)(t=3.870,P<0.01) and there was a higher number of MC present in patients with IC (5.59±3.40/HP) compared with controls (2.48±2.23/HP)(t=2.787,P<0.01).The unique finding that only MC express ER(R=0.884, P<0.01.3. URO-OVA mice bladders were collected after i.v.injection of activated OT-I T cells.Clear histological bladder inflammation was observed at days7(score,3) and14(score,2). These mast cells of URO-OVA mice were increased to12.95%after stimulation, no change for the URO-OVA/KitW-SH group after stimulation.Conclusion1. Mast cell infiltration may be one of the characteristic pathological manifestations of the interstitial cystitis; The association of mast cell infiltration and the PUF scale maybe be a new diagnosis criteria for interstitial cystitis.2. MC high affinity estrogen receptors may help to explain why interstitial cystitis occurs primarily in women,why IC and other associated symptoms tend to be worst at the midcycle. These results have shown that estrogen augment MC histamine secretion.3. URO-OVA mice spontaneously develop OVA-mediated autoimmune cystitis at10wk of age. The inflamed bladders showed phenotypic characteristics of chronic bladder inflammation like IC. There was virtually no change for the URO-OVA/KitW-SH group after stimulation.