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高膽紅素血癥及膽紅素腦病新生大鼠模型的建立及評(píng)價(jià)

發(fā)布時(shí)間:2018-06-15 23:19

  本文選題:高膽紅素血癥 + 膽紅素腦病。 參考:《山西醫(yī)科大學(xué)》2017年碩士論文


【摘要】:目的:建立可行可靠、步驟簡(jiǎn)單且具有良好模擬性的高膽紅素血癥及膽紅素腦病的7日齡SD大鼠模型,為在臨床實(shí)踐中探究高膽紅素血癥及膽紅素腦病的發(fā)病機(jī)制及并發(fā)癥的防治提供模型基礎(chǔ)。方法:選取7日齡的SD(Sprague-Dawley,SD)大鼠50只隨機(jī)分成5組,分別給予生理鹽水,50mg/kg、100mg/kg、150mg/kg、200mg/kg晶體膽紅素腹腔內(nèi)注射。在實(shí)驗(yàn)處理24h后取大鼠腦組織及心臟取血,測(cè)定血清、腦組織的膽紅素水平及S-100β濃度,進(jìn)行腦組織HE染色觀察海馬區(qū)神經(jīng)細(xì)胞的病理改變,用TUNEL法測(cè)定神經(jīng)細(xì)胞凋亡數(shù),計(jì)算凋亡率(Apoptotic index,AI)并及時(shí)記錄各組各指標(biāo)的實(shí)驗(yàn)結(jié)果,計(jì)算相關(guān)數(shù)據(jù)并對(duì)其進(jìn)行統(tǒng)計(jì)學(xué)分析,得出結(jié)論。進(jìn)一步評(píng)價(jià)新生大鼠模型的可靠性及穩(wěn)定性。結(jié)果:(1)觀察實(shí)驗(yàn)動(dòng)物一般情況:實(shí)驗(yàn)前,各組大鼠一般情況良好,皮膚顏色紅潤(rùn),彈性好,對(duì)外界刺激反應(yīng)靈敏,活動(dòng)正常。干預(yù)24小時(shí)后觀察:生理鹽水組的大鼠皮膚顏色紅潤(rùn),對(duì)外界刺激反應(yīng)靈敏;50mg/kg組的大鼠皮膚欠紅潤(rùn),彈性較差,對(duì)外界刺激反應(yīng)遲緩;100mg/kg組與150 mg/kg組新生大鼠的皮膚顏色黃染,彈性差,對(duì)外界刺激反應(yīng)遲鈍,有肌張力降低所致的翻滾及俯伏;200 mg/kg組的大鼠皮膚呈黃綠色,彈性極差,對(duì)外界刺激無(wú)反應(yīng)甚至昏迷。(2)HE染色:生理鹽水組海馬區(qū)神經(jīng)細(xì)胞排列整齊,結(jié)構(gòu)完整,形態(tài)正常;50mg/kg組可見神經(jīng)細(xì)胞之間輕度的腫脹、變性;100mg/kg組與150mg/kg組神經(jīng)元結(jié)構(gòu)紊亂,細(xì)胞之間腫脹、變形明顯,部分細(xì)胞核染色質(zhì)致密濃縮,胞漿空化;200mg/kg組神經(jīng)細(xì)胞之間極度腫脹、變形,可見多數(shù)細(xì)胞核染色質(zhì)致密濃縮,胞漿空化。(3)血清及腦組織膽紅素水平及S-100β濃度:與生理鹽水組相比,各組的血清及腦組織膽紅素水平及S-100β濃度均有統(tǒng)計(jì)學(xué)差異(P0.05);與50mg/kg組相比,100mg/kg組血清及腦組織的S-100β無(wú)顯著性差異(P0.05)。(4)TUNEL法測(cè)凋亡指數(shù)(AI):與生理鹽水組相比,各組的凋亡指數(shù)均有統(tǒng)計(jì)學(xué)差異(P0.05);與150mg/kg組相比,200mg/kg組凋亡指數(shù)無(wú)顯著性差異(P0.05)。結(jié)論:(1)7d SD大鼠腹腔注射50 mg/kg晶體膽紅素可建立高膽紅素血癥動(dòng)物模型。(2)7d SD大鼠腹腔注射100mg/kg與150 mg/kg晶體膽紅素可建立穩(wěn)定的膽紅素腦病模型。(3)當(dāng)腹腔注射150 mg/kg晶體膽紅素時(shí),神經(jīng)膠質(zhì)細(xì)胞分解明顯,所以7d SD大鼠注射150 mg/kg晶體膽紅素可建立膠質(zhì)細(xì)胞損傷指標(biāo)的模型。
[Abstract]:Objective: to establish a 7 day old SD rat model of hyperbilirubinemia and bilirubin encephalopathy. To provide a model for exploring the pathogenesis of hyperbilirubinemia and bilirubin encephalopathy and the prevention and treatment of complications in clinical practice. Methods: fifty SD rats aged 7 days were randomly divided into 5 groups, and were injected intraperitoneally with normal saline 50 mg / kg 100 mg / kg and 200 mg / kg intraperitoneal injection of intraperitoneal bilirubin. After 24 hours of experimental treatment, blood samples were taken from the brain and heart of rats, bilirubin level and S-100 尾 concentration in serum and brain tissue were measured, pathological changes of hippocampal neurons were observed by HE staining, apoptosis of neurons was measured by Tunel method. The apoptotic rate was calculated and the experimental results of each index were recorded in time. The relevant data were calculated and statistically analyzed, and the conclusion was reached. To further evaluate the reliability and stability of neonatal rat model. Results 1) observe the general situation of experimental animals: before the experiment, the rats in each group were generally in good condition, the skin color was ruddy, elastic, sensitive to external stimuli, normal activity. After 24 hours of intervention, the skin color of the normal saline group was ruddy, the skin of the 50 mg / kg group was less ruddy and the elasticity was poor, and the skin color of the newborn rats in the 100mg / kg group and the 150 mg/kg group were yellowish. The skin of the rats in the 200 mg/kg group was yellowish green and had extremely poor elasticity. No response to external stimuli or even coma with HE staining: the hippocampal neurons in the normal saline group were arranged neatly, the structure was intact, and slight swelling was observed between the neurons in the 50 mg / kg group of normal morphology, and the neuronal structure was disordered in the 100 mg / kg group and the 150mg/kg group. The cells were swollen and deformed, some of the nuclear chromatin was dense and concentrated, and the cytoplasmic cavitation of 200 mg / kg group was extremely swollen and deformed, and most of the nuclear chromatin were dense and concentrated. Serum and brain bilirubin level and S-100 尾 concentration: compared with normal saline group, The serum and brain bilirubin levels and S-100 尾 concentrations were significantly different in each group (P 0.05), and the S-100 尾 levels in serum and brain tissues were not significantly different from those in 50mg/kg group. The apoptotic index of each group was significantly different from that of 150mg/kg group (P 0.05), but there was no significant difference in apoptotic index between 200 mg / kg group and 150mg/kg group (P 0.05). Conclusion the hyperbilirubinemia animal model can be established by intraperitoneal injection of 50 mg/kg intraperitoneal bilirubin in 1 / 7 day SD rats with 1: 1% mg/kg and stable bilirubin encephalopathy model with intraperitoneal injection of 100mg/kg and 150 mg/kg intraperitoneal bilirubin) when intraperitoneal injection of bilirubin in 150 mg/kg crystal can be used to establish a stable model of bilirubin encephalopathy. The glial cells were decomposed obviously, so the injury index model of glial cells could be established by injecting 150 mg/kg crystal bilirubin into SD rats on day 7.
【學(xué)位授予單位】:山西醫(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)】:R722.1;R-332

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