【摘要】：1直腸癌新輔助放化療后放療相關(guān)性結(jié)腸纖維化的多光子光學(xué)診斷研究目的:探討多光子光學(xué)診斷技術(shù)在直腸癌輔助放化療后所致的放療相關(guān)性結(jié)腸纖維化的診斷作用。材料與方法:回顧性收集我科12例因放射性盆腔軟組織纖維化致腸管嚴(yán)重狹窄或腸梗阻患者的手術(shù)石蠟標(biāo)本(纖維化組)以及另外12例直腸癌新輔助放化療后隨訪期間未發(fā)生明顯纖維化的患者的手術(shù)石蠟標(biāo)本(未纖維化組),石蠟標(biāo)本取腸管近切端,以反映吻合口近端腸管的情況。另收集10例直腸癌新輔助放化療后患者(放化療組)以及10例未行新輔助放化療患者(未放化療組)的新鮮近切端手術(shù)標(biāo)本。應(yīng)用多光子光學(xué)診斷技術(shù)檢測實(shí)驗(yàn)樣本中的膠原密度、膠原纖維方向性指數(shù),同時應(yīng)用HE染色及VG染色的技術(shù)進(jìn)行對比研究。結(jié)果:多光子光學(xué)診斷技術(shù)可很好地顯示實(shí)驗(yàn)樣本中的膠原纖維及彈性纖維。放療可導(dǎo)致粘膜下層及固有肌層內(nèi)的膠原密度增高或出現(xiàn)膠原纖維破壞并出現(xiàn)大量不成熟的新生膠原纖維、粘膜下層內(nèi)膠原纖維排列紊亂以及彈性纖維的斷裂。石蠟標(biāo)本中兩組的粘膜下層內(nèi)膠原密度含量均高于肌層內(nèi)的膠原密度(p0.0001),纖維化組粘膜下層內(nèi)的膠原密度高于未纖維化組(p=0.0016),纖維化組肌層內(nèi)的膠原密度也高于未纖維化組(p0.0001)。纖維化組內(nèi)的膠原纖維方向性指數(shù)顯著高于未纖維化組(p0.0001)。新鮮標(biāo)本中兩組的粘膜下層內(nèi)膠原密度含量均高于肌層內(nèi)的膠原密度(p0.0001),放化療組粘膜下層內(nèi)的膠原密度高于未放化療組(p=0.0002),放化療組肌層內(nèi)的膠原密度也高于未放化療組(p=0.0014)。放化療組內(nèi)的膠原纖維方向性指數(shù)顯著高于未放化療組(p=0.0068)。結(jié)論:多光子光學(xué)診斷技術(shù)可以檢測放療后結(jié)腸腸管內(nèi)的早期纖維化相關(guān)病變。2吡非尼酮干預(yù)大鼠盆腔纖維化放療模型的實(shí)驗(yàn)研究目的:探討吡非尼酮干預(yù)大鼠盆腔纖維化放療模型中受照射腸管及腸管周圍肌肉組織纖維化的作用。材料與方法:應(yīng)用SD雄性大鼠構(gòu)建盆腔放射性纖維化的大鼠模型,并應(yīng)用吡非尼酮進(jìn)行干預(yù)研究。大鼠隨機(jī)分為空白對照組(A組)、吡非尼酮干預(yù)組(B組)及放射對照組(C組),在放射后10周每組大鼠取材5只,分別取大鼠直腸及直腸周圍肌肉組織進(jìn)行實(shí)驗(yàn)。應(yīng)用多光子活檢技術(shù)、HE染色及VG染色分析大鼠樣品內(nèi)膠原密度及膠原纖維方向性,RT-PCR及Western檢測TGF-β1、Smad3、CTGF等指標(biāo)的表達(dá)情況。結(jié)果:實(shí)驗(yàn)成功構(gòu)建大鼠放射性纖維模型。A組及B組中的粘膜下層膠原密度均顯著高于肌層內(nèi)的膠原密度(p均小于0.0001),而A組及B組間粘膜下層的膠原密度含量無顯著性差異(p=0.20),但C組粘膜下層膠原密度含量較B組則顯著性下降(p=0.004)。C組中可見肌層內(nèi)的膠原密度含量顯著性升高(p=0.0008)。三組方向性指數(shù)單因素方差分析比較具有顯著性差異(p=0.04)。三組大鼠直腸周圍肌肉內(nèi)的膠原纖維比例分別為0.024±0.002、0.08±0.003和0.14±0.006,單因素方差分析顯示三組之間存在顯著性差異(p0.0001);熒光定量PCR結(jié)果顯示三組的直腸(p=0.04)及直腸周圍肌肉(p=0.004)內(nèi)TGF-β1均存在顯著性差異,B組內(nèi)TGF-β1顯著下降。Smad3的表達(dá)水平在三組見未見顯著性差異。單因素方差分析顯示CTGF的表達(dá)水平在三組間未見統(tǒng)計學(xué)差異,但可見C組的表達(dá)水平有高于B組的趨勢;Western blot檢測結(jié)果顯示C組直腸組織中的TGF-β1水平顯著升高(p=0.036);但三組直腸周圍肌肉組織內(nèi)的TGF-β1水平差異無顯著性。Smad3表達(dá)在三組的直腸及其周圍肌肉均未見顯著性差異。CTGF水平在B組中的直腸及其周圍肌肉內(nèi)的表達(dá)均有顯著性下降。結(jié)論:吡非尼酮可減輕大鼠盆腔放射模型中的腸管及直腸周圍肌肉的纖維化,其可能是通過抑制組織內(nèi)TGF-β1及CTGF的蛋白質(zhì)表達(dá)起作用,對放射性相關(guān)纖維化的形成具有預(yù)防作用。多光子活檢技術(shù)可有效應(yīng)用于判斷大鼠盆腔放療模型中直腸腸管內(nèi)粘膜下層及肌層的纖維化情況。3吡非尼酮干預(yù)放療相關(guān)盆腔纖維化動物模型的基因表達(dá)譜分析目的:分析吡非尼酮對放療后盆腔纖維化大鼠模型基因表達(dá)譜的影響。材料與方法:應(yīng)用Affymetrix大鼠Genome 230 2.0 Array芯片對比分析空白對照組(A組)、吡非尼酮干預(yù)組(B組)及放射對照組(C組)大鼠的直腸組織內(nèi)基因表達(dá)譜變化情況。結(jié)果:B組與A組基因表達(dá)譜相比發(fā)差異表達(dá)基因的功能主要是對脂類的反應(yīng)、有機(jī)環(huán)化合物的反應(yīng)、類固醇類激素的反應(yīng)。C組與A組相比,顯著差異基因可以富集到的GO功能與胞外基質(zhì)相關(guān)的排序靠前,說明C組中胞外基質(zhì)受影響的程度比較明顯。B組與C組之間的差異表達(dá)基因GO功能富集到胞外空間、胞外基質(zhì)、細(xì)胞遷移等方面。三組之間相比所富集到的GO功能顯著不同。吡非尼酮干預(yù)后的通路富集與放射對照組顯著的不同。結(jié)論:三組大鼠直腸標(biāo)本的基因芯片表達(dá)譜對比分析表明,吡非尼酮具有減輕放射線對直腸組織細(xì)胞外基質(zhì)影響的作用,提示吡非尼酮具有預(yù)防放療相關(guān)纖維化的作用。
[Abstract]:1. Multiphoton Optical Diagnosis of Radiotherapy-associated Colon Fibrosis after Neoadjuvant Radiotherapy and Chemotherapy for Rectal Cancer Objective: To investigate the diagnostic value of multiphoton optic diagnostic technique in radiotherapy-associated Colon Fibrosis after Neoadjuvant Radiotherapy and Chemotherapy for Rectal Cancer. Surgical paraffin specimens from patients with severe stricture or intestinal obstruction (fibrosis group) and 12 patients with rectal cancer who did not have obvious fibrosis during the follow-up period after neoadjuvant radiotherapy and chemotherapy (non-fibrosis group) were taken from the proximal intestinal canal to reflect the situation of the proximal intestinal canal anastomosis. Fresh proximal resection specimens were obtained from adjuvant radiotherapy and chemotherapy group and 10 patients without neoadjuvant radiotherapy and chemotherapy group. Collagen density and directional index of collagen fibers in the experimental specimens were measured by multiphoton optical diagnostic technique. The collagen fibers and elastic fibers in the experimental samples can be well displayed by the sub-optical diagnostic technique. Radiotherapy can lead to increased collagen density in the submucosa and lamina propria or collagen fibers destruction and a large number of immature new collagen fibers, disorder of collagen fibers arrangement in the submucosa and breakage of elastic fibers. The collagen density in the submucosa of the fibrosis group was higher than that of the non-fibrosis group (p = 0.0016). The collagen density in the fibrosis group was higher than that in the non-fibrosis group (p 0.0001). The collagen Directional Index in the fibrosis group was significantly higher than that in the non-fibrosis group (p 0.0001). In the fresh samples, the collagen density in the submucosa was higher than that in the myometrium (p0.0001), the collagen density in the submucosa in the radiochemotherapy group was higher than that in the non-radiochemotherapy group (p = 0.0002), and the collagen density in the myometrium in the radiochemotherapy group was also higher than that in the non-radiochemotherapy group (p = 0.0014). The directional index was significantly higher than that of the control group (p = 0.0068). CONCLUSION: Multiphoton optical diagnostic technique can be used to detect the early fibrosis-related lesions in the colon and intestine after radiotherapy. 2 Experimental study of pirfenidone on the pelvic fibrosis radiotherapy model in rats Materials and Methods: SD male rats were used to establish the model of pelvic radiation fibrosis, and the intervention study was carried out with pirfenidone. The rats were randomly divided into blank control group (group A), pirfenidone intervention group (group B) and radiation control group (group C). The rats were taken from each group 10 weeks after radiation. Multiphoton biopsy, HE staining and VG staining were used to analyze the density and orientation of collagen fibers in rats. RT-PCR and Western blotting were used to detect the expression of TGF-beta 1, Smad3 and CTGF. The density of collagen in the submucosa of group B was significantly higher than that in the myometrium (p < 0.0001), but there was no significant difference in the density of collagen between group A and group B (p = 0.20), but the density of collagen in the submucosa of group C was significantly lower than that in group B (p = 0.004). (p = 0.0008). There was a significant difference among the three groups by one-way ANOVA (p = 0.04). The percentage of collagen fibers in the rectal muscles of the three groups was 0.024 [0.002], 0.08 [0.003] and 0.14 [0.006], respectively. There was a significant difference among the three groups by one-way ANOVA (p0.0001). The results of fluorescence quantitative PCR showed that there were significant differences among the three groups. TGF-beta 1 in rectum (p=0.04) and perirectal muscles (p=0.004) were significantly different, and TGF-beta 1 in group B was significantly decreased. Smad3 expression level was not significantly different among the three groups. The results of blot showed that the levels of TGF-beta 1 in rectal tissues of group C were significantly higher than those of group B (p=0.036), but there was no significant difference among the three groups. CONCLUSION: Pirfenidone can alleviate the fibrosis of intestinal canal and perirectal muscles in pelvic radiation model rats, which may play a role by inhibiting the expression of TGF-beta 1 and CTGF proteins in tissues and prevent the formation of radiation-related fibrosis. Gene expression profiling analysis of pirfenidone in radiotherapy-related pelvic fibrosis animal models Objective: To analyze the effect of pirfenidone on gene expression profiles in a rat model of pelvic fibrosis after radiotherapy. Materials and Methods: Genome 2302.0 Array microarray was used in Affymetrix rats. Results: The function of differentially expressed genes in group B was mainly to respond to lipids, organocyclic compounds and steroids compared with group A. The GO function enriched by the differentially expressed genes between group B and group C was enriched in extracellular space, extracellular matrix and cell migration. The GO function enriched by the differentially expressed genes between group B and group C was not significantly different from that enriched by the three groups. CONCLUSION: The comparative analysis of gene chip expression profiles of rectal specimens from three groups of rats shows that pirfenidone can alleviate the effect of radiation on extracellular matrix of rectal tissue, suggesting that pirfenidone can prevent radiation-related fibrosis.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R735.37;R730.55

【相似文獻(xiàn)】

中國期刊全文數(shù)據(jù)庫 前1條

1 陳勝;徐友志;杜偉;劉筆鋒;;基于多光子激發(fā)熒光-毛細(xì)管電泳的DNA分析新技術(shù)[J];光子學(xué)報;2008年02期

中國重要會議論文全文數(shù)據(jù)庫 前1條

1 劉立新;;多參量多焦點(diǎn)多光子顯微成像技術(shù)[A];2012年西部光子學(xué)學(xué)術(shù)會議論文摘要集[C];2012年

中國博士學(xué)位論文全文數(shù)據(jù)庫 前3條

1 吳燕;基于多光子顯微技術(shù)的乳腺腫瘤早期識別的研究[D];福建師范大學(xué);2016年

2 陳勝;毛細(xì)管電泳—連續(xù)光多光子激發(fā)熒光檢測系統(tǒng)的研制和應(yīng)用研究[D];華中科技大學(xué);2007年

3 劉嫩容;結(jié)直腸與食道癌組織的多光子顯微成像和拉曼光譜研究[D];福建師范大學(xué);2014年

中國碩士學(xué)位論文全文數(shù)據(jù)庫 前8條

1 周毅;基于多光子顯微成像技術(shù)橫縱切胃腸肌層的成像研究[D];福建師范大學(xué);2016年

2 梁閏富;1700nm波段多光子成像中信號優(yōu)化及溫升問題研究[D];深圳大學(xué);2016年

3 陳義拴;SiPM光子探測性能分析及高精度多光子探測研究[D];陜西師范大學(xué);2016年

4 占成;多光子隨機(jī)掃描顯微鏡的光學(xué)設(shè)計與構(gòu)建[D];華中科技大學(xué);2006年

5 李洪生;多光子顯微成像技術(shù)用于結(jié)直腸腺癌發(fā)展進(jìn)程監(jiān)測的研究[D];福建師范大學(xué);2014年

6 鐘家照;多光子顯微技術(shù)用于人體組織診斷的研究[D];福建師范大學(xué);2012年

7 劉玉春;多光子成像技術(shù)用于腸癌的早期診斷[D];福建師范大學(xué);2012年

8 李明;SO_2分子多光子激發(fā)光譜[D];河北大學(xué);2013年

，

本文編號：2214309