本文關(guān)鍵詞：中華眼鏡蛇毒對(duì)大鼠炎性細(xì)胞因子影響的研究　出處：《廣西醫(yī)科大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 中華眼鏡蛇毒 腫瘤壞死因子-α 白細(xì)胞介素-6 大鼠 作用機(jī)制

【摘要】：目的：通過(guò)腹腔內(nèi)注射中華眼鏡蛇毒(CCV)建立眼鏡蛇傷大鼠模型,以檢測(cè)血清及心肌組織中腫瘤壞死因子-α(TNF-α)和白細(xì)胞介素-6(IL-6)的表達(dá),從而探討炎性細(xì)胞因子在CCV所致大鼠眼鏡蛇傷中的致炎機(jī)制以及心肌損傷的機(jī)制。 方法：60只健康清潔級(jí)Sprague-Dawley (SD)大鼠在廣西醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心飼養(yǎng)3天后開始實(shí)驗(yàn)。SD大鼠被隨機(jī)(隨機(jī)數(shù)字法)等分為四組,即：對(duì)照組、染毒1h組、染毒3h組及染毒6h組。將CCV凍干粉按lmg/m1與生理鹽水配成蛇毒溶液,染毒各組大鼠予以腹腔內(nèi)注射1.272mg/kg CCV溶液；對(duì)照組大鼠給予等劑量生理鹽水溶液腹腔內(nèi)注射。 (1)染毒后,連續(xù)觀察各組大鼠中毒表現(xiàn)直至實(shí)驗(yàn)終止； (2)染毒各組大鼠于染毒后相應(yīng)時(shí)間點(diǎn)即染毒1h、3h、6h行腹主動(dòng)脈采血2.0 ml,對(duì)照組大鼠于1h后同法采血,標(biāo)本在室溫下靜置20分鐘后離心(3000r/min,10min),收集血清,采用酶聯(lián)免疫吸附法(ELISA法)測(cè)定血清TNF-a和IL-6抗原含量,以探討炎性細(xì)胞因子TNF-αIL-6在CCV所致大鼠炎癥反應(yīng)中的致炎機(jī)制； (3)同時(shí),染毒各組大鼠于染毒后各相應(yīng)時(shí)間點(diǎn)切取心肌組織,行HE染色；對(duì)照組大鼠于觀察1h后同法切取心肌組織,通過(guò)光學(xué)顯微鏡觀察心肌組織結(jié)構(gòu)的變化； (4)按上述方法,染毒各組和對(duì)照組分別切取心肌組織,采用免疫組織化學(xué)方法檢測(cè)心肌組織中TNF-α及工L-6的表達(dá),進(jìn)行圖像分析并計(jì)算其平均灰度值,研究染毒后大鼠心肌組織中炎性細(xì)胞因子TNF-α和IL-6的表達(dá)變化。 結(jié)果： (1)腹腔內(nèi)注射CCV溶液后,染毒各組大鼠逐漸出現(xiàn)中毒反應(yīng),而對(duì)照組大鼠在整個(gè)實(shí)驗(yàn)觀察期間未出現(xiàn)明顯的中毒癥狀； (2)與對(duì)照組比較,大鼠血清TNF-a、IL-6抗原含量在染毒各組中表達(dá)增高：血清TNF-α含量在染毒1h組開始增加,染毒6h組明顯增加；血清工L-6含量在染毒1h組增加不明顯,染毒3h組開始增加,染毒6h組增加明顯； (3)光學(xué)顯微鏡下觀察染毒6h組大鼠心肌結(jié)構(gòu)出現(xiàn)明顯的病理改變,主要表現(xiàn)為心肌纖維出現(xiàn)部分?jǐn)嗔选⒛[脹,間質(zhì)水腫,出現(xiàn)炎性細(xì)胞的浸潤(rùn)；對(duì)照組大鼠心肌結(jié)構(gòu)無(wú)明顯改變。 (4)本實(shí)驗(yàn)結(jié)果顯示,與對(duì)照組比較,染毒1、3、6h大鼠心肌細(xì)胞內(nèi)可見明顯TNF-α和工L-6陽(yáng)性表達(dá)細(xì)胞,呈棕黃色顆粒狀,分布于細(xì)胞漿內(nèi),且陽(yáng)性細(xì)胞平均灰度值明顯降低。結(jié)論： (1)本研究采用腹腔內(nèi)注射CCV溶液的方法建造眼鏡蛇傷大鼠模型,在造模后染毒各組大鼠均出現(xiàn)相應(yīng)的中毒表現(xiàn)以及心肌組織結(jié)構(gòu)的變化,表明大鼠染毒模型建造成功。 (2)染毒各組大鼠血清TNF-α和工L-6抗原含量明顯增加,表明炎性細(xì)胞因子在CCV所致大鼠眼鏡蛇傷炎癥反應(yīng)發(fā)生機(jī)制中有促炎作用。 (3)本研究表明CCV能夠使大鼠心肌組織中TNF-α和工L-6表達(dá)明顯增加以及心肌組織結(jié)構(gòu)形態(tài)發(fā)生改變,提示TNF-α、IL-6在大鼠眼鏡蛇傷心肌損傷發(fā)生機(jī)制中可能起到重要作用。
[Abstract]:Objective: through intraperitoneal injection of Chinese cobra venom (CCV) to establish the cobra injury rat model, to detect serum and myocardial tissue tumor necrosis factor alpha (TNF- alpha) and interleukin -6 (IL-6) so as to explore the expression of inflammatory cytokines in Cobra CCV rats induced by injury of the inflammatory mechanism of myocardial injury and the mechanism.
Methods: 60 healthy Sprague-Dawley rats (SD) in the animal experimental center of Guangxi Medical University feeding after 3 days of experimental.SD rats were randomly (random number) divided into four groups: control group, 1H exposure group, 3h group and 6h group were exposed. The CCV freeze-dried powder and saline by lmg/m1 with venom solution, rats exposed to intraperitoneal injection of 1.272mg/kg CCV solution; the control group rats were given saline solution intraperitoneal injection.
(1) after the poisoning, the poisoned performance of the rats in each group was continuously observed until the end of the experiment.
(2) in rats after exposure to the corresponding time point is at 1H, 3h, 6h for abdominal aortic blood 2 ml, rats in the control group after 1h with the method of blood samples at room temperature and set aside for 20 minutes after centrifugation (3000r/min, 10min), serum was collected by ELISA (ELISA) method for determination of serum TNF-a and IL-6 antigen content, to investigate the inflammatory cytokines TNF- induced by CCV IL-6 in the inflammatory response in rats with inflammatory mechanism;
(3) at the same time, exposure of rats after exposure to the corresponding time points in myocardial tissues by HE staining; the control group rats were observed after 1h with Fletcher myocardium, changes of myocardial tissue structure was observed by optical microscope;
(4) according to the above method, exposure and control group were harvested for myocardial tissue, expression of myocardial tissue were detected by immunohistochemical method in TNF- alpha and L-6, image analysis and calculation of the average gray value of inflammatory cells after exposure in myocardium of rats with expression of TNF- alpha and IL-6.
Result錛，

本文編號(hào)：1403279