本文選題：椎間盤　切入點(diǎn)：糖尿病　出處：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:測定鏈脲佐菌素處理后造成糖尿病狀態(tài)的大鼠和正常大鼠椎間盤組織中轉(zhuǎn)化生長因子β(TGF-β)和胰島素樣生長因子-1(IGF-1)在三個(gè)不同時(shí)期的含量變化,并探討含量之間差異的意義。方法:1 3月齡成熟雄性SD大鼠30只,隨機(jī)分為實(shí)驗(yàn)組以及對(duì)照組,每組SD大鼠各15只,實(shí)驗(yàn)組SD大鼠采用腹腔單次注射鏈脲佐菌素溶液(strep-tozotoein,STZ),通過鏈脲佐菌素對(duì)胰腺胰島β-細(xì)胞的制毒作用,致使胰島素生成障礙,從而造成糖尿病狀態(tài),建立糖尿病大鼠模型,對(duì)照組SD大鼠不做特殊處理,僅給予腹腔單次注射檸檬酸鹽緩沖液。2分別在第2、4、6周時(shí),從實(shí)驗(yàn)組和對(duì)照組各取5只大鼠,用1%戊巴比妥鈉進(jìn)行腹腔注射處死大鼠后,取其各組SD大鼠新鮮腰椎間盤組織,用4%多聚甲醛進(jìn)行固定,梯度乙醇脫水,包埋成石蠟組織塊。分別行蘇木素-伊紅(HE)染色法,二甲苯透明后,中性樹脂封片顯微鏡下觀察兩組大鼠椎間盤組織病理變化情況。免疫組化法,脫蠟加入一抗二抗后,辣根酶標(biāo)記,DAB顯色顯微鏡下觀察兩組大鼠椎間盤組織中TGF-β和IGF-1的含量。選取其中最有意義的,陽性表達(dá)最強(qiáng)的5個(gè)視野進(jìn)行計(jì)算,以細(xì)胞質(zhì)著色和陽性細(xì)胞率為判斷標(biāo)準(zhǔn),統(tǒng)計(jì)兩組大鼠椎間盤中TGF-β和IGF-1的表達(dá)情況。結(jié)果:1經(jīng)鏈脲佐菌素注射液處理后的糖尿病組模型大鼠,血糖濃度明顯升高,與正常組對(duì)比,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。糖尿病組血糖濃度分別在第2、4、6周時(shí)相對(duì)于正常組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。2在3個(gè)不同時(shí)期,實(shí)驗(yàn)組大鼠的椎間盤均出現(xiàn)不同程度的退行性改變,HE染色顯示可見脊索細(xì)胞減少,增生出較多膠原纖維,終板呈不同程度的鈣化,出現(xiàn)類軟骨細(xì)胞,纖維環(huán)呈蜿蜒狀排列,髓核基質(zhì)排列紊亂、減少,呈不同程度的皺縮,髓核及纖維環(huán)向外膨出;部分髓核細(xì)胞排列成簇,這是椎間盤退變的標(biāo)志。而對(duì)照組正常大鼠,椎間盤結(jié)構(gòu)正常,終板由一層透明軟骨組成,纖維環(huán)具有規(guī)則的層狀結(jié)構(gòu),膠原束排列整齊平行,以成纖維細(xì)胞為主,髓核與纖維環(huán)分界清楚,其中成條索狀分布的脊索細(xì)胞較為豐富,基質(zhì)中細(xì)胞充實(shí)。3免疫組化法測定,對(duì)照組大鼠中TGF-β染色陽性率在3個(gè)不同時(shí)期無差異(P=0.368),對(duì)照組大鼠中IGF-1染色陽性率在3個(gè)不同時(shí)期無差異(P=0.368);實(shí)驗(yàn)組大鼠中TGF-β的陽性染色率在3個(gè)不同時(shí)期無差異(P=0.311),實(shí)驗(yàn)組大鼠中IGF-1的陽性染色率在3個(gè)不同時(shí)期無差異(P=0.727)。實(shí)驗(yàn)組大鼠椎間盤組織中TGF-β的含量在3個(gè)不同時(shí)期均明顯低于對(duì)照組(P0.05)。實(shí)驗(yàn)組大鼠椎間盤組織中IGF-1的含量在3個(gè)不同時(shí)期均明顯低于對(duì)照組(P0.05)。結(jié)論:本研究結(jié)果顯示實(shí)驗(yàn)組中STZ誘導(dǎo)的糖尿病大鼠椎間盤中TGF-β和IGF-1含量較對(duì)照組正常大鼠明顯減少,在椎間盤HE染色中可以觀察到,實(shí)驗(yàn)組糖尿病大鼠椎間盤的形態(tài)發(fā)生了退變性改變,這種TGF-β和IGF-1含量的降低,引起了椎間盤內(nèi)基質(zhì)的降解,抑制軟骨基質(zhì)的合成,使其未分化間充質(zhì)細(xì)胞的分化減退,加速了椎間盤內(nèi)的膠原水解,降低椎間盤內(nèi)II型膠原和蛋白多糖等軟骨細(xì)胞表型的能力,最終使椎間盤因?yàn)槿鄙僮銐虻幕|(zhì)水分而發(fā)生彈性下降、結(jié)構(gòu)紊亂、穩(wěn)定性及代謝生理功能異常等改變,導(dǎo)致椎間盤退變。本研究結(jié)果證明了糖尿病使椎間盤內(nèi)TGF-β和IGF-1的含量發(fā)生降低,這可能是導(dǎo)致糖尿病患者椎間盤發(fā)生退行性改變的原因之一。這為今后糖尿病患者預(yù)防、治療椎間盤發(fā)生的退行性改變提供了新的治療思路。
[Abstract]:Objective: to determine the streptozotocin treated by diabetes status and normal rat intervertebral disc tissue transforming growth factor beta (TGF- beta) and insulin-like growth factor -1 (IGF-1) in three different periods of the content changes, and to explore the significance of the difference between the content. Method: 13 month old adult male 30 SD rats were randomly divided into experimental group and control group, each group of SD rats, 15 rats in each experimental group, SD rats by a single intraperitoneal injection of streptozotocin solution (strep-tozotoein, STZ), on pancreatic islet beta cell precursor by STZ, resulting in insulin production obstacles, resulting in the diabetic state, establish the model of diabetic rats, the control group of SD rats did not do special treatment, only received a single intraperitoneal injection of citrate buffer.2 respectively in 2,4,6 weeks, from each experimental group and control group 5 rats, abdominal with 1% pentobarbital sodium The rats were sacrificed after injection, the group of SD rats fresh lumbar intervertebral disc tissue was fixed with 4% paraformaldehyde, dehydrated in graded ethanol and embedded into paraffin blocks respectively. Hematoxylin and eosin (HE) staining, xylene transparent, observe two groups of rat intervertebral disc tissue the change of neutral resin mounting microscope. Immunohistochemical method, adding a two anti anti dewaxing, HRP, DAB color microscope TGF- P and IGF-1 two groups of rat intervertebral disc tissues. The most significant, the 5 strongest vision positive expression was calculated by cytoplasmic staining and positive cell rate of the expression of statistical criteria, two groups of rats intervertebral discs TGF- beta and IGF-1. Results: 1 the diabetic group rats by streptozotocin injection after treatment, blood glucose concentration increased significantly, compared with the normal group, the difference was statistically significant (P0.05). The blood glucose concentration in the diabetic group were 2,4,6 weeks when compared with normal group, the difference was statistically significant (P0.05.2) in 3 different periods, the intervertebral disc of the experimental rats showed varying degrees of degenerative change, HE staining showed that the notochord cells decreased, more collagen fiber hyperplasia, endplate was different degrees of calcification, like cartilage cells, the fiber ring is arranged in the form of winding, nucleus pulposus disorder, reduce, showed different degrees of shrinkage, nucleus pulposus and annulus bulge outward; part of the nucleus pulposus cells arranged in clusters, which is a sign of intervertebral disc degeneration. While the control group of normal rats, intervertebral disc structure normally, a layer of transparent cartilage endplate, layered annulus has rules, collagen bundles arranged parallel to fibroblasts, nucleus pulposus and annulus clear boundaries, which become a notochord cell cord distribution is abundant, medium In full.3 cells were determined by immunohistochemical method, the rats in the control group TGF- beta staining was no difference in 3 different periods (P=0.368), IGF-1 control group rats in the positive rate had no difference in 3 different periods (P=0.368); the positive staining of experimental rats in TGF- beta rate the differences in the 3 different periods (P=0.311), the positive staining of IGF-1 in experimental group rats was no difference in the 3 different periods (P=0.727). The content of the experimental group of rat intervertebral disc tissue of TGF- beta in 3 different periods were significantly lower than the control group (P0.05). The experimental group of rat intervertebral disc the fabric in the content of IGF-1 in 3 different periods were significantly lower than the control group (P0.05). Conclusion: the results of this study showed that the experimental group in STZ induced diabetic rats intervertebral discs TGF- beta and IGF-1 content compared with the control group of normal rats was significantly reduced, the intervertebral disc HE staining can be observed in the experimental group The morphogenesis of the intervertebral disc degenerative change, reduce the TGF- beta and IGF-1 content, caused the degradation of intervertebral disc matrix, inhibit the synthesis of cartilage matrix, the differentiation of undifferentiated mesenchymal cells decreased, accelerates the hydrolysis of collagen in the intervertebral disc, reduce the ability of phenotype intradiscal and collagen II protein sugar of cartilage cells, the intervertebral disc because of the lack of sufficient and elastic matrix moisture decreased, structure disorder, metabolic stability and physiological function abnormal, intervertebral disc degeneration. The results of this study proved that the content of diabetes intradiscal TGF- beta and IGF-1 incidence decreased, which may lead to intervertebral in patients with diabetes mellitus one of the causes of disc degeneration. This is the future of diabetes prevention, provide a new treatment strategy for degenerative intervertebral disc changes.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R587.2;R681.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 Neeraj Kumar Agrawal;Saket Kant;;Targeting inflammation in diabetes: Newer therapeutic options[J];World Journal of Diabetes;2014年05期

2 王凱;張亮;張強(qiáng);周濤;姜竹巖;胡金海;劉鳳松;;腰椎軟骨終板細(xì)胞凋亡與椎間盤退變的關(guān)系[J];天津醫(yī)科大學(xué)學(xué)報(bào);2012年01期

3 陳森;徐振華;賈吉光;張蕾;鄭曉佐;曹平;;糖尿病大鼠終板微血管改變與髓核細(xì)胞凋亡的關(guān)系[J];武漢大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年05期

4 ;IL-1beta sensitizes rat intervertebral disc cells to Fas ligand mediated apoptosis in vitro[J];Acta Pharmacologica Sinica;2007年10期

5 鮑恒,侯筱魁;糖尿病大鼠椎間盤細(xì)胞凋亡的觀察[J];中華骨科雜志;2005年07期

6 彭寶淦,侯樹勛,賈連順,陳德玉;頸椎軟骨終板鈣化與頸椎間盤退變和椎體骨贅形成的關(guān)系[J];中國脊柱脊髓雜志;2001年06期

7 彭寶淦,賈連順,施杞,沈培芝;軟骨終板鈣化在椎間盤退變過程中的作用機(jī)理[J];中國矯形外科雜志;2000年02期

相關(guān)碩士學(xué)位論文 前1條

1 何勇;糖尿病對(duì)腰椎間盤退變的影響[D];浙江大學(xué);2010年

，

本文編號(hào)：1710326