發(fā)布時(shí)間：2018-06-06 02:46

  本文選題：鎘 + 姬松茸多糖�。� 參考：《東北農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：鎘是一種有害重金屬物質(zhì),可造成人和動(dòng)物機(jī)體外周血液淋巴細(xì)胞(Peripheral blood lymphocytes,PBLs)和腎臟毒性損傷,破壞自身免疫系統(tǒng)。姬松茸多糖(Agaricus blazei Murill Polysaccharides,ABP)能夠治療鎘帶來的毒性損傷。本試驗(yàn)從細(xì)胞凋亡水平、模式識別受體TLR4信號通路及抗氧化角度研究ABP對鎘中毒雞PBLs和腎臟的影響,以明確ABP對鎘中毒雞的保護(hù)作用。將7日齡生長狀態(tài)良好的雛雞隨機(jī)分為四組,每組20只。鎘組飼喂含氯化鎘140 mg/kg的全價(jià)飼料,多糖組飲食為全價(jià)飼料并給予0.2 m L ABP,糖鎘組飼喂含氯化鎘140 mg/kg的全價(jià)飼料,ABP 0.2 m L,對照組雞以正常飼養(yǎng)方式使用全價(jià)飼料飼喂。分別在第20、40、60試驗(yàn)日時(shí),心臟采取雞外周血液,分離、收集外周血液淋巴細(xì)胞,細(xì)胞培養(yǎng)24 h后收集細(xì)胞上清,并采取雞腎臟,通過電感耦合等離子-質(zhì)譜法(IPC-MS)檢測雞PBLs及腎臟鎘含量;流式細(xì)胞術(shù)測定并計(jì)算雞PBLs凋亡率;實(shí)時(shí)熒光定量PCR法對雞PBLs凋亡基因Caspase-3、Bax和Bcl-2 m RNA,PBLs和腎臟TLR4及TLR4信號通路相關(guān)因子(My D88、TRIF、NF-κB、IRF3)mRNA水平進(jìn)行檢測;Western blot方法分析TLR4蛋白變化;雙抗體夾心ELISA法檢測PBLs和腎臟炎癥因子IL-1β、IL-6、TNF-α的含量;通過MDA、T-SOD、GSH-p X測試盒對雞腎臟中相應(yīng)因子進(jìn)行檢測。結(jié)果發(fā)現(xiàn),鎘組雞PBLs和腎臟組織中的鎘含量顯著高于對照組,ABP治療后其鎘含量明顯下降。鎘組雞PBLs細(xì)胞凋亡率最高,ABP治療后凋亡率顯著抑制,同時(shí)多糖組細(xì)胞凋亡率明顯低于對照組。同對照組相比,鎘組雞PBLs中Caspase-3、Bax的mRNA水平升高,Bcl-2 mRNA水平降低,Bax/Bcl-2明顯升高,ABP干預(yù)后,上述變化得到顯著抑制。ABP能顯著抑制鎘誘導(dǎo)的雞PBLs和腎臟TLR4表達(dá)以及My D88、TRIF、NF-κB、IRF3 m RNA水平。降低鎘組雞炎癥因子IL-1β、IL-6、TNF-α含量。鎘組雞MDA含量顯著高于對照組,而T-SOD、GSH-p X的活性明顯降低,姬松茸多糖進(jìn)行治療后得到明顯改善,且多糖組MDA的含量與對照組相比顯著降低,T-SOD、GSH-p X的活性顯著升高。上述結(jié)果表明,鎘能夠誘導(dǎo)雞外周血液淋巴細(xì)胞凋亡,活化雞PBLs和腎臟組織TLR4信號通路,增加炎癥因子的表達(dá)釋放,損害雞腎臟氧化-抗氧化功能,造成雞PBLs和腎臟損傷,ABP對鎘引起的上述損傷均有明顯的保護(hù)作用。綜上所述,本試驗(yàn)通過建立雞鎘中毒模型,研究鎘對雞PBLs和腎臟毒性損傷作用以及姬松茸多糖對鎘毒性的影響。為今后進(jìn)一步研究鎘毒性及ABP的治療機(jī)制提供理論依據(jù)。
[Abstract]:Cadmium is a harmful heavy metal substance, which can damage peripheral blood lymphocytes PBLs and kidney toxicity in human and animal peripheral blood, and destroy the autoimmune system. Agaricus blazei Murill polysaccharide ABP can treat the toxic injury caused by cadmium. The effects of ABP on PBLs and kidney of cadmium poisoned chickens were studied in terms of cell apoptosis level, pattern recognition receptor TLR4 signaling pathway and antioxidation angle, in order to clarify the protective effect of ABP on cadmium intoxicated chickens. The 7-day-old chicks were randomly divided into four groups with 20 chickens in each group. The cadmium group was fed with the full price diet containing cadmium chloride 140 mg/kg, the polysaccharide group was fed the full price diet and the 0.2 m L ABP, glucose cadmium group was fed with ABP 0.2 mL containing cadmium chloride 140 mg/kg, while the control group was fed with the full price diet in the normal way. On the 20th day of the experiment, chicken peripheral blood was isolated, lymphocytes were collected, supernatants were collected after 24 hours of cell culture, and chicken kidneys were taken. The contents of cadmium in PBLs and kidney were detected by inductively coupled plasma-mass spectrometry (ICP-MS), and the apoptosis rate of chicken PBLs was measured and calculated by flow cytometry. The changes of TLR4 protein in chicken PBLs apoptosis gene Caspase-3 Bax and Bcl-2 m RNA-PBLs and renal TLR4 and TLR4 signal transduction related factors were detected by real-time fluorescence quantitative PCR. The changes of TLR4 protein were analyzed by Western blot, and the contents of PBLs and the renal inflammatory factor IL-1 尾 -IL-6TNF- 偽 were detected by double antibody sandwich ELISA method. The corresponding factors in chicken kidney were detected by MDA-T-SODX GSH-p X test box. The results showed that the content of cadmium in PBLs and renal tissue in cadmium group was significantly higher than that in control group after treatment. The apoptosis rate of PBLs cells in cadmium group was the highest and the apoptosis rate in polysaccharide group was significantly lower than that in control group. Compared with the control group, the mRNA level of Caspase-3 and Bax in PBLs of cadmium group was increased and the level of Bcl-2 mRNA was decreased. The above changes were significantly inhibited. ABP could significantly inhibit the expression of PBLs and renal TLR4 induced by cadmium, and the level of NF- 魏 IRF3 m RNA of my D88 trifion (NF- 魏 BIF-IRF3). The content of IL-1 尾 -IL-6 TNF- 偽 was decreased in cadmium group. The content of MDA in cadmium group was significantly higher than that in control group, while the activity of T-SODN GSH-pX was decreased significantly, and the content of MDA in polysaccharide group was significantly improved after treatment, and the activity of GSH-pX in polysaccharide group was significantly lower than that in control group. These results suggest that cadmium can induce apoptosis of lymphocytes in chicken peripheral blood, activate TLR4 signaling pathway in chicken PBLs and kidney tissue, increase the expression and release of inflammatory factors, and damage the oxidation-antioxidation function of chicken kidney. Both PBLs and ABP have obvious protective effects on cadmium-induced injury. To sum up, the effects of cadmium on PBLs and kidney toxicity and the effects of polysaccharides of Agaricus blazei Murrill on cadmium toxicity were studied. It provides a theoretical basis for the further study of cadmium toxicity and the therapeutic mechanism of ABP.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S858.31

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