本文關(guān)鍵詞： 泮托拉唑 腫瘤惡病質(zhì) 炎癥 泛素蛋白酶體途徑　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的本實(shí)驗(yàn)通過(guò)建立腫瘤惡病質(zhì)小鼠模型,研究泮托拉唑?qū)翰≠|(zhì)狀態(tài)下的骨骼肌消耗的保護(hù)作用及其可能的相關(guān)機(jī)制,為惡病質(zhì)的進(jìn)一步研究提供新的思路,并為臨床應(yīng)用提供理論依據(jù)。方法48只雄性BALB/c小鼠,按照隨機(jī)數(shù)字表分為6個(gè)組,每組8只,分別為:正常對(duì)照組(NC)、惡病質(zhì)組(CC)、低濃度治療組(CL)、中濃度治療組(CM)、高濃度治療組(CH)、生理鹽水組(CS)。荷瘤小鼠于右前肢腋下接種小鼠結(jié)腸腺癌細(xì)胞c26,以建立小鼠腫瘤惡病質(zhì)模型;正常對(duì)照組于相同部位注射等量的PBS緩沖液。每日監(jiān)測(cè)小鼠的體重、腫瘤的大小以及各組小鼠的自發(fā)活動(dòng)。待接種腫瘤細(xì)胞的小鼠進(jìn)入到惡病質(zhì)狀態(tài)后,治療組采用不同濃度的泮托拉唑灌胃,生理鹽水組采用等量生理鹽水灌胃,NC組和CC組不作任何干預(yù)。待藥物治療2周后,收集標(biāo)本檢測(cè)相關(guān)指標(biāo)。ELISA法檢測(cè)血清IL-6及TNF-α水平。Real-time PCR檢測(cè)腓腸肌JAK2及STAT3 m RNA表達(dá)情況;Western blot檢測(cè)腓腸肌p-JAK2,p-STAT3,Fbx32和Mu RF1蛋白表達(dá)情況。免疫組化法檢測(cè)腓腸肌組織中Fbx32和Mu RF1的表達(dá)。HE染色觀察腓腸肌形態(tài)變化。采用SPSS17.0軟件分析數(shù)據(jù)。結(jié)果1腫瘤惡病質(zhì)模型成功建立荷瘤小鼠在接種腫瘤細(xì)胞后第5天,皮下可觸及腫瘤結(jié)節(jié)生長(zhǎng);接種后第12天,荷瘤小鼠腫瘤明顯增大,體重較正常對(duì)照組明顯下降(p0.05),并且出現(xiàn)了自發(fā)性活動(dòng)減少(p0.05)、毛發(fā)暗淡無(wú)光、精神虛弱等癥狀,提示荷瘤小鼠進(jìn)入惡病質(zhì)狀態(tài)。2小鼠體質(zhì)量及腓腸肌變化接種腫瘤細(xì)胞后第5天,荷瘤小鼠皮下可觸及腫瘤結(jié)節(jié),但各組小鼠體重?zé)o差異(p0.05);接種c26細(xì)胞后第12天,荷瘤小鼠較正常組體重明顯減輕(p0.05);在泮托拉唑干預(yù)2周后,治療組小鼠較生理鹽水組體重有不同程度增加,且高濃度組小鼠體重增加最為明顯(p0.05)。實(shí)驗(yàn)結(jié)束時(shí),治療組小鼠較生理鹽水組腓腸肌重量及橫切面積均有不同程度增加,且高濃度組小鼠體重增加最為明顯(p0.05);高濃度組小鼠的腫瘤重量較生理鹽水組也有明顯減輕(p0.05)。3小鼠血清中炎性因子水平變化與正常對(duì)照組相比,血清炎性因子IL-6和TNF-α水平在惡病質(zhì)小鼠中顯著升高(p0.05)。在使用泮托拉唑干預(yù)后,各濃度組較生理鹽水組血清炎性因子都有不同程度降低,尤其高濃度組降低得最為顯著(p0.05)。4小鼠炎性通路JAK2/STAT3表達(dá)變化與正常對(duì)照組相比,炎性通路JAK2和STAT3的m RNA表達(dá)水平在惡病質(zhì)小鼠中顯著升高(p0.05),并且p-JAK2和p-STAT3的蛋白表達(dá)水平在惡病質(zhì)小鼠中也明顯增高(p0.05)。在使用泮托拉唑干預(yù)后,各濃度治療組同生理鹽水組比較,JAK2和STAT3的m RNA表達(dá)水平以及p-JAK2和p-STAT3的蛋白表達(dá)水平都有不同程度降低,尤其以高濃度組降低最為顯著(p0.05)。5小鼠骨骼肌泛素蛋白酶體系統(tǒng)變化與正常對(duì)照組比較,惡病質(zhì)組小鼠腓腸肌中的Mu RF1和Fbx32的蛋白表達(dá)水平明顯升高(p0.05),而泮托拉唑干預(yù)后,不同濃度干預(yù)組小鼠的Mu RF1和Fbx32的蛋白表達(dá)水平較生理鹽水組均降低,且高濃度組降低最為明顯(p0.05)。結(jié)論小鼠C26細(xì)胞接種于BALB/c小鼠成功建立腫瘤惡病質(zhì)模型。泮托拉唑能明顯改善腫瘤惡病質(zhì)一般情況,增加體質(zhì)量,緩解骨骼肌萎縮,抑制腫瘤生長(zhǎng),并且高濃度泮托拉唑具有更好的保護(hù)作用。泮托拉唑緩解腫瘤惡病質(zhì)骨骼肌消耗可能是通過(guò)降低炎性因子水平,抑制炎性通路JAK2/STAT3的激活,抑制泛素蛋白酶體途徑。
[Abstract]:Objective to establish a mouse model of cancer cachexia by this experiment, the protective effect of pantoprazole on the consumption state of the skeletal muscle cachexia and related mechanism, to provide new ideas for the further research of cachexia, and provide theoretical basis for clinical application. Methods: 48 male BALB/c mice were randomly divided into 6 group, 8 rats in each group, respectively: normal control group (NC), cachexia group (CC), low dose treatment group (CL), the concentration of the treatment group (CM), high dose treatment group (CH), normal saline group (CS). The tumor bearing mice inoculated with mouse colon to the right forelimb adenocarcinoma cell C26, in order to establish the model of cancer cachexia mice; PBS buffer in normal control group was injected with the same amount of daily monitoring. The weight of mice, the tumor size and the mice spontaneous activity. After inoculation of tumor cells into mice to cachexia state, The treatment group was treated with different concentration of pantoprazole orally, saline group with normal saline, NC group and CC group without any intervention. 2 weeks after drug treatment, samples were collected to detect the serum IL-6 and TNF- level.Real-time.ELISA index method to detect PCR JAK2 and STAT3 m of gastrocnemius muscle RNA expression Western; blot detection of gastrocnemius p-JAK2 p-STAT3, Fbx32 Mu and RF1 protein expression. To observe the morphological changes of.HE Fbx32 expression of gastrocnemius muscle and Mu RF1 were detected by immunohistochemistry staining in gastrocnemius tissues. Data analysis by SPSS17.0 software. Results of the 1 tumor cachexia model was successfully established in mice after inoculation of tumor cells the fifth day, subcutaneous tumor nodules were palpable; twelfth days after inoculation, the tumor bearing mice significantly increased, weight decreased significantly compared with the control group (P0.05), and the emergence of spontaneous activity reduction (P0.05), hair Dim and dark, mental weakness and other symptoms, suggesting that cachexia.2 mice body weight and gastrocnemius muscle changes fifth days after inoculation of tumor cells into nude mice bearing subcutaneous tumor bearing mice, palpable tumor nodules, but no difference in body weight of mice in each group (P0.05); twelfth days after inoculation of C26 cells, the tumor bearing mice compared with the normal group significant weight loss (P0.05); after 2 weeks of intervention in pantoprazole, treatment group mice compared with saline group weight increased to different degrees, and the weight of the high concentration group mice increased significantly (P0.05). At the end of the experiment, mice in treatment group compared with the saline group and the gastrocnemius muscle weight cutting area increased in varying degrees, and weight the high concentration group mice increased significantly (P0.05); high concentration group mice tumor weight compared with the saline group was significantly reduced (P0.05) changes in the levels of inflammatory factors in serum of.3 mice compared with the normal control group, serum inflammatory Factor IL-6 and TNF- levels increased significantly in cachexia in mice (P0.05). In the use of pantoprazole intervention groups compared with saline group serum inflammatory factors were decreased in different degree, especially in the high concentration group decreased most significantly (P0.05).4 mice JAK2/ STAT3 expression in the inflammatory pathway and normal compared with the control group, the expression level of M RNA and STAT3 JAK2 inflammatory pathway increased significantly in cachexia in mice (P0.05), and the expression of p-JAK2 and p-STAT3 protein levels were significantly higher in cachexia in mice (P0.05). In the use of pantoprazole after the intervention, the treatment group was with normal saline group, m RNA JAK2 and STAT3 and the expression of p-JAK2 and p-STAT3 protein expression levels were decreased in different degree, especially in the high concentration group was reduced significantly (P0.05) in skeletal muscle of.5 mice ubiquitin proteasome system change compared with normal control group, The expression level of cachexia mice gastrocnemius in Mu RF1 and Fbx32 protein increased significantly (P0.05), and pantoprazole intervention, different concentrations of Mu and RF1 in the intervention group Fbx32 protein expression levels compared with the saline group decreased, and decreased in high concentration group was most significant (P0.05). Conclusion C26 in mice the cells were inoculated into BALB/c mice successfully established model of cancer cachexia. Pantoprazole can significantly improve tumor cachexia in general, increased body weight, alleviate skeletal muscle atrophy, inhibit tumor growth, protective effect and high concentration of pantoprazole has better. Pantoprazole reduced the consumption of cancer cachexia skeletal muscle is likely to decrease levels of inflammatory factors the activation, inhibition of inflammatory pathways of JAK2/STAT3, inhibition of the ubiquitin proteasome pathway.

【學(xué)位授予單位】：重慶醫(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)】：R730.5

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