【摘要】：目的 研究肥大細胞（Mast cell, MC）、神經(jīng)纖維（nerve fiber）、Toll樣受體3（Toll-like receptor3, TLR3）及Toll樣受體4（Toll-like receptor4, TLR4）在先天性腸無神經(jīng)節(jié)細胞癥（Hirschsprung’s disease, HD）患者不同腸段中分布和表達情況，探究其分布和表達差異的臨床意義。 方法 選擇我院病理診斷為HD的手術(shù)切除結(jié)腸標(biāo)本40例，另取尸檢結(jié)直腸標(biāo)本10例（心血管疾病死亡9例和狂犬病死亡1例）作為對照組。將40例實驗組標(biāo)本的痙攣段、移行段、擴張段及10例對照組腸段標(biāo)本，采用HE染色、甲苯胺藍化學(xué)染色、S-P免疫組織化學(xué)染色法及復(fù)合染色法進行處理，觀察肥大細胞、神經(jīng)纖維、TLR3、TLR4的分布表達及形態(tài)學(xué)變化情況，采用彩色病理圖像分析軟件進行半定量分析，運用SPSS17.0進行統(tǒng)計學(xué)處理。 結(jié)果 ①鏡下肥大細胞呈圓形、梭形或不規(guī)則形，胞漿呈藍紫色，胞核淺藍色或不染色，主要集中在腸段黏膜層及黏膜下層，與血管或神經(jīng)纖維毗鄰，HD患者痙攣段肥大細胞較對照組及擴張段明顯增多（P㩳0.05），脫顆粒現(xiàn)象明顯。 ②S-100免疫反應(yīng)陽性的神經(jīng)纖維呈黃棕色顆粒，位于黏膜下神經(jīng)叢及肌間神經(jīng)叢中，呈散在的點狀、線狀或簇集分布。在HD患者痙攣段黏膜下層可見S-100陽性反應(yīng)的神經(jīng)纖維增生明顯，染色增強，呈波浪形，，面積較對照組明顯增大（P0.05）。肥大細胞密度與神經(jīng)纖維的密度（r=0.573，P0.01）及面積（r=0.514，P0.01）存在正相關(guān)性。 ③TLR3定位于細胞胞質(zhì)中，主要表達在粘膜層的上皮細胞、神經(jīng)叢內(nèi)的神經(jīng)節(jié)細胞及神經(jīng)纖維，對平均光密度分析比較，各個腸段中粘膜層中的TLR3表達無顯著統(tǒng)計學(xué)差異（P0.05），肌間神經(jīng)叢內(nèi)TLR3表達存在統(tǒng)計學(xué)差異（P 0.05），從正常組到病變段遞減。 ④TLR4主要在腸道肌層高表達，而在神經(jīng)叢不表達或弱表達，HD痙攣段肌層內(nèi)TLR4表達量與對照組及擴張段比較，明顯增高(P0.05)。 結(jié)論 肥大細胞與神經(jīng)纖維關(guān)系密切，兩者的變化及相互作用在HD的發(fā)病中可能具有重要意義，肥大細胞數(shù)量變化及脫顆�，F(xiàn)象在腸無神經(jīng)節(jié)細胞癥的診斷具有一定意義；TLR3、TLR4在HD不同腸段的表達分別具有差異性，兩者在腸無神經(jīng)節(jié)細胞癥的發(fā)病中可能起著一定作用，TLR3可以作為病理診斷巨結(jié)腸的標(biāo)記物之一。
[Abstract]:Objective to study the role of (Mast cell, MC), nerve fiber (nerve fiber), Toll like receptor 3 (Toll-like receptor3, TLR3) and Toll like receptor 4 (Toll-like receptor4, TLR4) in the diagnosis of (Hirschsprung's disease, in congenital intestinal apogoneurosis. To explore the clinical significance of the distribution and expression of HD in different intestinal segments. Methods Forty surgically resected colon specimens diagnosed as HD and 10 autopsy colorectal specimens (9 died of cardiovascular disease and 1 death of rabies) were selected as control group. HE staining, toluidine blue staining, S-P immunohistochemical staining and compound staining were used to observe mast cells in 40 cases of spastic segment, transitional segment, dilated segment and 10 cases of control group, which were treated by HE staining, toluidine blue staining, S-P immunohistochemical staining and compound staining. The distribution, expression and morphologic changes of nerve fibers and TLR3,TLR4 were analyzed by color pathological image analysis software and statistically analyzed by SPSS17.0. Results 1Mast cells were round, fusiform or irregular, cytoplasm was blue-purple, nucleus was light blue or unstained, mainly concentrated in intestinal mucosa and submucous layer, adjacent to blood vessels or nerve fibers. The mast cells in spastic segment of HD patients were more than those in control group and dilated segment (P0. 05). The 2S-100 immunoreactive nerve fibers were yellowish brown, located in the submucous plexus and intermuscular plexus, and distributed in scattered dots, lines or clusters. S-100 positive nerve fibers in the submucosa of the spastic segment of HD patients were obviously proliferated, stained and wave-shaped, and the area was significantly larger than that of the control group (P0.05). There was a positive correlation between the density of mast cells and the density of nerve fibers (r ~ (0.573) P _ (0.01) and the area (r ~ (0.5 14) P _ (0.01). 3TLR3 was localized in cytoplasm and mainly expressed in epithelial cells in mucosal layer, ganglion cells and nerve fibers in plexus. The mean optical density was analyzed and compared. There was no significant difference in the expression of TLR3 in the mucosal layer of each intestinal segment (P0.05), but the expression of TLR3 in the myenteric plexus was significantly different (P 0.05), which decreased from normal group to pathological segment. The expression of 4TLR4 was mainly high in the intestinal myometrium, but not or weakly expressed in the nerve plexus. The expression of TLR4 in the muscular layer of HD spastic segment was significantly higher than that in the control group and the dilated segment (P0.05). Conclusion Mast cells are closely related to nerve fibers. The changes and interactions between mast cells and nerve fibers may play an important role in the pathogenesis of HD. The expression of TLR3,TLR4 in different intestinal segments of HD may play a role in the pathogenesis of AGC. TLR3 can be used as a marker for pathological diagnosis of Hirschsprung's disease.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R726.5

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