本文選題：乳桿菌 + 肥胖�。� 參考：《哈爾濱工業(yè)大學(xué)》2017年碩士論文

【摘要】：高脂膳食不僅能導(dǎo)致超重,還會誘發(fā)骨代謝和吸收的異常,導(dǎo)致骨量減少和骨強度降低,從而導(dǎo)致自發(fā)和創(chuàng)傷性骨損傷的風(fēng)險增加,嚴重危害骨健康。骨代謝異�；蚬俏帐芏喾矫嬉蛩赜绊�,其中營養(yǎng)膳食因素占據(jù)重要地位。但是,長期高脂膳食與骨性能降低的關(guān)系目前并未有確切解釋。益生菌對骨健康狀態(tài)的改善主要是通過對免疫系統(tǒng)的調(diào)控來影響破骨細胞的活性,達到調(diào)節(jié)骨吸收與骨重建平衡的作用。因此,我們推測腸道菌群可能是治療緩解骨質(zhì)疏松的一個新靶點。但是益生菌與免疫系統(tǒng)及骨吸收之間具體的關(guān)系仍需進一步探討。本課題擬從兩株乳酸菌中篩選出對高脂膳食誘導(dǎo)Balb/c小鼠肥胖有明顯抑制作用的益生菌株,研究其對肥胖小鼠骨性能的影響;探究肥胖小鼠腸道菌群變化、結(jié)腸免疫因子及糞便短鏈脂肪酸的關(guān)系,進而推斷乳桿菌促進肥胖小鼠骨健康的機制。(1)采用灌胃的方式,將本實驗室分離鑒定得到的Lactobacillus paracasei subsp.Paracasei M5(M5)及Lactobacillus coryniformis subsp.torquens T3L(T3L)乳桿菌以109 CFU/mL的劑量給予造模成功的肥胖Balb/c小鼠,4周末,T3L、M5均顯著降低小鼠體重,體脂率及Lee’s指數(shù)(p0.05)。通過測定小鼠血清甘油三酯(TG)和總膽固醇(TC),結(jié)果表明M5及T3L均顯著降低血清TG、TC含量(p0.05)。通過對各組小鼠的肝臟進行HE染色切片發(fā)現(xiàn),經(jīng)T3L、M5乳桿菌干預(yù)后,高脂膳食小鼠肝臟脂肪病變的程度有所改善,脂肪空泡變小而且變少。結(jié)果顯示T3L菌株效果較顯著。綜上結(jié)果提示,M5及T3L菌株能抑制高脂飲食導(dǎo)致的小鼠肥胖及血脂上升。(2)乳桿菌對肥胖小鼠骨健康具有保護作用:測定小鼠血清ALP、TRAP及RANKL的含量,結(jié)果顯示灌胃T3L乳桿菌4周后,血清中這三者的水平得到顯著降低(p0.05),說明高脂膳食使得破骨細胞增生活躍,機體骨吸收增加,灌胃乳桿菌后得到改善。經(jīng)T3L乳桿菌干預(yù)后,肥胖小鼠股骨生物力學(xué)性能增強、骨礦含量增加,且通過Micro-CT掃描發(fā)現(xiàn)股骨微結(jié)構(gòu)均得到有效地改善。RT-qPCR法檢測小鼠股骨組織中相關(guān)因子的表達,結(jié)果顯示高脂膳食使股骨中RANKL和CTSK mRNA表達水平顯著上調(diào)(p0.05),經(jīng)T3L灌胃4周顯著降低了其表達量;同時,T3L乳桿菌的攝入顯著地提高了COL1α1的表達(p0.05)。(3)通過高通量測序法對小鼠腸道菌群多樣性進行分析。高脂膳食誘導(dǎo)的肥胖使小鼠腸道菌群出現(xiàn)差異,灌胃乳桿菌T3L后,小鼠腸道菌群的多樣性發(fā)生改變,產(chǎn)短鏈脂肪酸菌屬增多。通過RT-qPCR檢測結(jié)腸組織中IL-10、TGF-β1及Foxp3的表達,發(fā)現(xiàn)T3L的補充顯著上調(diào)了高脂膳食小鼠腸內(nèi)IL-10的表達,且表達水平接近對照組,p0.05。同時,灌胃T3L乳桿菌后糞便中短鏈脂肪酸的含量增多。益生菌可能通過調(diào)節(jié)腸道菌群結(jié)構(gòu),使產(chǎn)短鏈脂肪酸菌比例增加,從而利用短鏈脂肪酸影響結(jié)腸免疫功能進而干預(yù)骨重塑機制來完成對骨的保護作用。
[Abstract]:High fat diet can not only lead to overweight, but also induce abnormal metabolism and absorption of bone, reduce bone mass and bone strength, thus lead to increased risk of spontaneous and traumatic bone injury, and seriously endanger bone health. Abnormal bone metabolism or bone resorption is affected by many factors, among which nutritional and dietary factors play an important role. However, the relationship between long-term high-fat diet and bone loss has not been explained. The improvement of bone health by probiotics is mainly through the regulation of the immune system to affect the activity of osteoclasts and to regulate the balance between bone resorption and bone reconstruction. Therefore, we speculate that intestinal flora may be a new target for the treatment of osteoporosis. But the specific relationship between probiotics and immune system and bone resorption needs to be further explored. In this study, two strains of lactic acid bacteria were screened out to inhibit obesity induced by high fat diet in Balb/c mice, to study the effect of probiotics on the bone properties of obese mice, and to explore the changes of intestinal flora in obese mice. The relationship between colon immune factors and faecal short chain fatty acids, and then infer the mechanism of Lactobacillus to promote bone health in obese mice. Lactobacillus paracasei subsp.Paracasei M5 (M5) and Lactobacillus coryniformis subsp.torquens T3 LnT3L) were isolated and identified in our laboratory. Lactobacillus T3L5 and Lactobacillus coryniformis subsp.torquens T3 LnT3L) were given to obese Balb/c mice at the dose of 109 CFU/mL. The weight, body fat rate and Lee's index of the obese Balb/c mice were significantly decreased at the end of the 4th week. The results showed that both M5 and T3L significantly decreased the serum TGN TC content in mice. The liver sections of each group of mice were stained with HE staining. It was found that after the intervention of Lactobacillus T3L5, the degree of fatty lesions in liver of high-fat diet mice was improved, and the fat vacuoles became smaller and fewer. The results showed that the effect of strain T 3 L was significant. The results showed that strain M5 and T3L could inhibit obesity and increase of blood lipid in mice induced by high fat diet. Lactobacillus lactis had protective effect on bone health of obese mice. Serum levels of ALP trap and RANKL were measured. The results showed that Lactobacillus sp. T3L was perfused 4 weeks after oral administration of Lactobacillus t3L. The serum levels of these three factors were significantly decreased (P 0.05), which indicated that high fat diet made osteoclast proliferation active, bone resorption increased, and Lactobacillus gastric administration improved. After the intervention of Lactobacillus T3L, the biomechanical properties and bone mineral content in the femur of obese mice were enhanced, and the expression of related factors in the femur of obese mice was detected by RT-qPCR, and the microstructures of femur were all improved by Micro-CT scanning. The results showed that high fat diet significantly upregulated the expression of RANKL and CTSK mRNA in femur. At the same time, the uptake of Lactobacillus sp. T3L significantly increased the expression of COL1 偽 1. The intestinal flora of mice induced by high-fat diet was different. The diversity of intestinal flora of mice was changed and the short-chain fatty acid producing bacteria was increased after Lactic bacteria T3L was infused into the stomach. The expression of IL-10 TGF- 尾 1 and Foxp3 in colon tissue was detected by RT-qPCR. It was found that T3L supplementation significantly upregulated the expression of IL-10 in the intestine of high-fat diet mice, and the expression level was close to that of control group (p0.05. At the same time, the content of short chain fatty acids in feces increased after gastric administration of Lactobacillus. Probiotics may increase the proportion of short-chain fatty acid producing bacteria by adjusting the structure of intestinal flora, so that the short chain fatty acids can affect the immune function of colon and interfere with the mechanism of bone remodeling to protect the bone.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R580;TS201.3

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