【摘要】：[目的]研究氟化鈉對(duì)小鼠腸道大腸桿菌、乳酸桿菌和雙歧桿菌生長(zhǎng)情況、糖代謝酶和蛋白表達(dá)的影響。[方法]以小鼠腸道分離培養(yǎng)的大腸桿菌、乳酸桿菌和雙歧桿菌為試驗(yàn)菌株,在培養(yǎng)液中依次加入氟化鈉,使其濃度達(dá)到0.1mmol/L、1mmol/L、10mmol/L、 100mmol/L,振蕩培養(yǎng)。利用比濁法檢測(cè)大腸桿菌、乳酸桿菌和雙歧桿菌的生長(zhǎng)情況,采用酸度計(jì)檢測(cè)細(xì)菌的產(chǎn)酸情況。利用酶活性檢測(cè)試劑盒測(cè)定糖代謝關(guān)鍵酶的活性。提取大腸桿菌、乳酸桿菌和雙歧桿菌總蛋白,采用SDS-PAGE分析細(xì)菌蛋白的表達(dá)變化。[結(jié)果]1.在培養(yǎng)24h內(nèi),低濃度氟化鈉(0.1mmol/L、1mmol/L)對(duì)大腸桿菌的生長(zhǎng)有一定的促進(jìn)作用,同時(shí)抑制產(chǎn)酸、增加大腸桿菌的產(chǎn)氣;高濃度的氟化鈉(100mmol/L)對(duì)大腸桿菌的生長(zhǎng)有明顯的抑制作用,同時(shí)明顯地抑制產(chǎn)酸、減少大腸桿菌的產(chǎn)氣。在培養(yǎng)48h內(nèi),低濃度氟化鈉(0.1mmol/L)對(duì)乳酸桿菌的生長(zhǎng)、產(chǎn)酸無(wú)明顯促進(jìn)作用,對(duì)雙歧桿菌的生長(zhǎng)、產(chǎn)酸有促進(jìn)作用；高濃度氟化鈉(100mmol/L)對(duì)乳酸桿菌和雙歧桿菌的生長(zhǎng)、產(chǎn)酸有明顯抑制作用。2.氟化鈉濃度為10mmol/L時(shí),大腸桿菌糖酵解代謝過(guò)程的己糖激酶、丙酮酸激酶、琥珀酸脫氫酶和檸檬酸合酶的酶活值明顯偏低；氟化鈉濃度為1mmol/L時(shí),乳酸桿菌和雙歧桿菌的己糖激酶、丙酮酸激酶和乳酸脫氫酶的酶活值明顯偏低。3.氟化鈉濃度為10mmol/L作用18h,大腸桿菌蛋白濃度為0.33mg/mL,150kDa和25kDa處的蛋白表達(dá)上調(diào),45kDa和31kDa處的蛋白表達(dá)下調(diào)。氟化鈉濃度為1mmol/L作用36h,乳酸桿菌蛋白濃度為1.78mg/mL,250kDa、132kDa和75kDa處的蛋白表達(dá)上調(diào),92kDa,47kDa和31kDa處的蛋白表達(dá)下調(diào)。氟化鈉濃度為1mmol/L作用36h,雙歧桿菌蛋白濃度1.69mg/mL,219kDa、75kDa和28kDa處的蛋白表達(dá)上調(diào),62kDa、47kDa和31kDa處的蛋白表達(dá)下調(diào)。[結(jié)論]低濃度氟化鈉可促進(jìn)大腸桿菌的生長(zhǎng)和產(chǎn)氣,但抑制其產(chǎn)酸,同時(shí)會(huì)促進(jìn)雙歧桿菌的生長(zhǎng)和產(chǎn)酸,但對(duì)乳酸桿菌的生長(zhǎng)、產(chǎn)酸無(wú)明顯作用；高濃度氟化鈉可抑制大腸桿菌、乳酸桿菌和雙歧桿菌的生長(zhǎng)和產(chǎn)酸,同時(shí)會(huì)抑制大腸桿菌的產(chǎn)氣。氟化鈉濃度為10mmol/L時(shí),即可抑制大腸桿菌某些糖代謝酶的活性；氟化鈉濃度為1mmol/L時(shí),即可抑制乳酸桿菌和雙歧桿菌某些糖代謝酶的活性。氟化鈉會(huì)影響大腸桿菌、乳酸桿菌和雙歧桿菌蛋白的表達(dá)。
[Abstract]:[objective] to study the effects of sodium fluoride on the growth of Escherichia coli, Lactobacillus and bifidobacterium in mice and the expression of glucose metabolizing enzymes and proteins. [methods] Escherichia coli, Lactobacillus and bifidobacterium isolated from the intestinal tract of mice were used as test strains. Sodium fluoride was added into the culture medium in turn, and the concentration was 0.1 mmol / L, 1 mmol / L, 10 mmol / L, 100 mmol / L, respectively. The growth of Escherichia coli, Lactobacillus and bifidobacterium was detected by turbidimetric method, and the acid production of bacteria was detected by acidometer. Enzyme activity test kit was used to determine the activity of key enzymes in glucose metabolism. Total proteins of Escherichia coli, Lactobacillus and Bifidobacterium were extracted and analyzed by SDS-PAGE. [result] 1. Within 24 hours, low concentration of sodium fluoride (0.1 mmol / L, 1 mmol / L) promoted the growth of Escherichia coli, inhibited the acid production and increased the gas production of E. coli. High concentration of sodium fluoride (100mmol/L) can obviously inhibit the growth of Escherichia coli, at the same time, it can obviously inhibit the production of acid and reduce the gas production of E. coli. In 48 h, low concentration of sodium fluoride (0.1mmol/L) did not promote the growth of Lactobacillus and acid production, but promoted the growth and acid production of Bifidobacterium. High concentration of sodium fluoride (100mmol/L) significantly inhibited the growth and acid production of Lactobacillus and Bifidobacterium. The enzyme activities of hexokinase, pyruvate kinase, succinate dehydrogenase and citrate synthase in the glycolytic process of Escherichia coli were significantly lower when the concentration of sodium fluoride was 10mmol/L. The enzyme activities of hexokinase, pyruvate kinase and lactate dehydrogenase of Lactobacillus and Bifidobacterium were significantly lower when the concentration of sodium fluoride was 1mmol/L. When the concentration of sodium fluoride was 10mmol/L for 18 h, the protein concentration of E. coli was 0.33 mg / mL, the protein expression at 150kDa and 25kDa was up-regulated, and the protein expression at 45kDa and 31kDa was down-regulated. After exposure to sodium fluoride for 36 hours, the protein concentration of Lactobacillus was 1.78 mg / mL, 250 kDaa, 132 kDa and 75kDa was up-regulated, while the protein expression at 92 KD, 47 kDa and 31kDa was down-regulated. At the concentration of 1mmol/L for 36 h, the protein concentration of Bifidobacterium was 1.69 mg / mL, the protein expression at 219 KD, 75kDa and 28kDa was up-regulated, while the protein expression at 62kDa, 47kDa and 31kDa was down-regulated. [conclusion] Sodium fluoride at low concentration can promote the growth and gas production of Escherichia coli, but inhibit its acid production, and promote the growth and acid production of Bifidobacterium, but has no obvious effect on the growth of Lactobacillus. High concentration of sodium fluoride can inhibit the growth and acid production of Escherichia coli, Lactobacillus and bifidobacterium, and inhibit the gas production of Escherichia coli. When the concentration of sodium fluoride is 10mmol/L, the activities of some glucose metabolizing enzymes of Escherichia coli can be inhibited, and the activities of some glucose metabolizing enzymes of Lactobacillus and Bifidobacterium can be inhibited when the concentration of sodium fluoride is 1mmol/L. Sodium fluoride affects the expression of E. coli, Lactobacillus and bifidobacterium proteins.
【學(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S852.6

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