本文選題：草分枝桿菌　切入點(diǎn)：小鼠　出處：《吉林農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：草分枝桿菌(Mycobacterium phlei)是一種在自然界廣泛存在的天然藥物資源,含有多種免疫活性成分。在醫(yī)學(xué)領(lǐng)域中,草分枝桿菌注射劑烏體林斯已經(jīng)被廣泛應(yīng)用于紫癜、哮喘等多種疾病的治療,但是其應(yīng)用于免疫方面的研究很少,特別是在畜牧業(yè)生產(chǎn)中,草分枝桿菌應(yīng)用的報(bào)道非常罕見(jiàn),同時(shí)未見(jiàn)有比較不同處理方法制備的草分枝桿菌對(duì)動(dòng)物免疫功能影響的研究報(bào)道。為此,本研究主要探討不同方法獲得的草分枝桿菌制品對(duì)小鼠免疫功能的影響。首先隨機(jī)選取10只小鼠進(jìn)行急性毒性試驗(yàn),小鼠灌服草分枝桿菌菌液0.2mL/天/只,其中含有草分枝桿菌有效成分110mg。然后選取120只體重18-22g的昆明小鼠,隨機(jī)分為5組,試驗(yàn)組分別為草分枝桿菌活菌組、細(xì)胞壁組、熱滅活組、超聲破碎組,以生理鹽水為對(duì)照。每組24只,4個(gè)草分枝桿菌試驗(yàn)組分別灌服不同方法獲得的草分枝桿菌制品,劑量為110mg/只/d,灌胃體積為0.2mL,定期稱(chēng)量小鼠體重,6周后每組隨機(jī)選取6只小鼠,測(cè)定其胸腺指數(shù)和脾臟指數(shù),用抗凝血測(cè)定其血常規(guī)及部分肝腎功能指標(biāo);然后每個(gè)試驗(yàn)組再取6只小鼠取血,分離血清用比濁法測(cè)定血清中溶菌酶,用生物素雙抗體夾心酶聯(lián)免疫吸附法測(cè)定血清IL-2和IgG含量;無(wú)菌分離脾臟并制備脾細(xì)胞懸液用MTT法測(cè)定其淋巴細(xì)胞轉(zhuǎn)化率和NK細(xì)胞活性;再每組另取6只小鼠,測(cè)定其腹腔巨噬細(xì)胞吞噬酵母菌能力;每組最后剩余的6只小鼠攻鼠傷寒沙門(mén)菌,進(jìn)行免疫保護(hù)試驗(yàn)。結(jié)果表明:①急性毒性試驗(yàn)結(jié)果顯示:草分枝桿菌對(duì)試驗(yàn)小鼠無(wú)毒。②5組小鼠血常規(guī)指標(biāo)PLT、MPV、PDW和P-LCR以及肝腎功能指標(biāo)ALB、TCHE、UREA、CRE和TG水平均差異不顯著;而對(duì)于血常規(guī)中的WBC、RBC、HGB以及HCT等指標(biāo)和肝腎功能指標(biāo)中的ALP、TP以及ALT等指標(biāo)的水平,草分枝桿菌4個(gè)試驗(yàn)組均高于生理鹽水組,但是均在正常范圍內(nèi)。③草分枝桿菌4個(gè)試驗(yàn)組對(duì)小鼠體重的影響與對(duì)照組相比無(wú)顯著性差異。④對(duì)于小鼠的臟器指數(shù),草分枝桿菌4個(gè)試驗(yàn)組的脾臟指數(shù)和胸腺指數(shù)均顯著高于生理鹽水組,其中細(xì)胞壁組的脾臟指數(shù)顯著高于其余3組,但后三者之間差異不顯著;活菌組和細(xì)胞壁組的胸腺指數(shù)顯著高于熱滅活組和超聲破碎組,而后兩者差異不顯著。⑤草分枝桿菌4個(gè)試驗(yàn)組的淋巴細(xì)胞轉(zhuǎn)化率、NK細(xì)胞活性、巨噬細(xì)胞吞噬功能和血清IL-2含量均顯著高于生理鹽水組;其中活菌組和細(xì)胞壁組的刺激指數(shù)差異無(wú)統(tǒng)計(jì)學(xué)意義,但顯著高于熱滅活組和超聲破碎組,而后二者差異不顯著;細(xì)胞壁組和熱滅活組的NK細(xì)胞活性和IL-2含量差異不顯著,但較活菌組和超聲破碎組明顯提高,后二者之間差異無(wú)統(tǒng)計(jì)學(xué)意義;而對(duì)于草分枝桿菌4個(gè)試驗(yàn)組的巨噬細(xì)胞吞噬功能,細(xì)胞壁組顯著高于其余三組,活菌組和熱滅活組之間無(wú)顯著性差異,但顯著高于超聲破碎組。⑥草分枝桿菌4個(gè)試驗(yàn)組的血清中溶菌酶的活性均顯著高于生理鹽水組,熱滅活組的溶菌酶活性顯著高于活菌組、細(xì)胞壁組和超聲破碎組,但后三者之間差異不顯著;草分枝桿菌細(xì)胞壁組、熱滅活組的血清IgG含量顯著高于活菌組、超聲破碎組以及生理鹽水組,但后三者之間差異無(wú)統(tǒng)計(jì)學(xué)意義�！�7灌胃結(jié)束后,通過(guò)注射沙門(mén)菌進(jìn)行小鼠攻毒試驗(yàn),攻毒后7天,免疫保護(hù)試驗(yàn)結(jié)果顯示,草分枝桿菌活菌組、細(xì)胞壁組、熱滅活組、超聲破碎組和生理鹽水組對(duì)小鼠的攻毒保護(hù)率分別為33.3%、66.7%、66.7%、50%和0,表明草分枝桿菌能在一定程度上增強(qiáng)小鼠的細(xì)胞免疫和體液免疫功能。綜上說(shuō)明,草分枝桿菌是一種安全有效的免疫調(diào)節(jié)劑,能明顯增強(qiáng)小鼠的免疫能力。
[Abstract]:Mycobacteriumphlei (Mycobacterium phlei) is a kind of widely existed in the nature of natural medicine resources, containing a variety of immune activity components. In the field of medicine, mycobacteriumphlei Utilins injection has been widely used in the treatment of purpura, asthma and other diseases, but is rarely used in the study of the immunity, especially in animal husbandry in addition, mycobacteriumphlei application reports are very rare, and there has been no report about the comparison of different treatment methods of Mycobacterium phlei preparation effect on animal immune function. Therefore, this study aims to investigate the effect of Mycobacterium phlei products obtained by different methods on immune function of mice. The first 10 mice were randomly selected for acute toxicity test, mice fed mycobacteriumphlei bacteria 0.2mL/ / day, which contains the effective components of Mycobacterium phlei 110mg. then selected 120 weight 18-22g Kunming The mice were randomly divided into 5 groups, experimental group were mycobacteriumphlei bacteria group, cell wall components, heat inactivated group, ultrasonic group, physiological saline control group. Each group had 24 rats, 4 mycobacteriumphlei experimental groups were obtained by taking different methods of mycobacteriumphlei products, dose 110mg/ /d, intragastric volume was 0.2mL, the weight of mice after 6 weeks of regular weighing, each group of 6 mice were randomly selected, the determination of the thymus index and spleen index, the determination of blood with anticoagulant and part of liver and kidney function index; then each experimental group and 6 mice were separated from blood serum by determination of serum lysozyme turbidimetric method, determination of serum IL-2 and IgG content with biotin double antibody sandwich enzyme-linked immunosorbent assay; sterile isolated spleen and preparation of spleen cell determination of conversion rate and NK cell activity in the cell suspension with MTT method; then another 6 mice in each group, the determination of peritoneal macrophages Cell phagocytosis of yeast; 6 mice in each of the last remaining Salmonella typhimurium, immune protection test. The results showed that: the results of acute toxicity test showed that Mycobacterium phlei mice non-toxic. In the 5 groups of Mice Blood Routine Indexes of PLT, MPV, PDW and P-LCR as well as liver and kidney function indexes of ALB, TCHE, UREA. CRE and TG were no significant difference in blood; and for WBC, RBC, HGB, HCT and other indicators of liver and kidney function indexes of ALP, TP, ALT and other indicators of the level, mycobacteriumphlei 4 test groups were higher than that of saline group, but were within the normal range. 3 mycobacteriumphlei 4 a group of test of influence on body weight of mice compared with the control group had no significant difference. 4 for the organ index of mice, mycobacteriumphlei 4 in experimental group, spleen index and thymus index were significantly higher than those in the saline group, the cell wall group spleen index significantly Compared with the other 3 groups, but no significant difference between the three groups; the thymus index of live bacteria and cell wall group was significantly higher than that of heat inactivated group and ultrasonic group, and there was no significant difference between them. 5 mycobacteriumphlei 4 in experimental group, lymphocyte transformation rate, NK cell activity, the function and the content of serum IL-2 phagocytosis of macrophages were significantly higher than those in the saline group; there was no significant bacterial cell wall group group and the stimulation index difference, but was significantly higher than that of heat inactivated group and ultrasonic group, and the difference between the two is not significant; the difference of NK cell activity and IL-2 content of cell wall components and heat inactivated group is not obviously, but compared with live bacteria group and ultrasonic group increased significantly, there was no significant difference between the two; and for mycobacteriumphlei 4 test group of macrophage phagocytosis, cell wall group was significantly higher than the other three groups, between live bacteria group and heat inactivated group A significant difference, but significantly higher than ultrasonic group. The serum lysozyme mycobacteriumphlei 4 test groups in the activity were significantly higher than those in the saline group, heat inactivated group of lysozyme activity was significantly higher than that of live bacteria, cell wall group and ultrasonic group, but no significant difference between the latter three Mycobacterium; coli cell wall components, heat inactivated serum content of IgG group was significantly higher than that of live bacteria group, ultrasonication group and normal saline group, but there was no significant difference between the three. 7 hours after gavage, mice were challenged by Salmonella test injection, 7 days after infection, the immune protection test results Mycobacterium phlei, living bacteria group, cell wall components, heat inactivated group, ultrasonic group and saline group of mice infected protection rates were 33.3%, 66.7%, 66.7%, 50% and 0, showed that Mycobacterium phlei can enhance mouse cell immunity and body to a certain extent In conclusion, Mycobacterium Mycobacterium is a safe and effective immunomodulator, which can obviously enhance the immune ability of mice.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S859.79
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本文編號(hào)：1570823