本文關(guān)鍵詞：鐵皮石斛水提物對(duì)糖尿病小鼠血糖和腸道菌群的影響　出處：《大連醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 鐵皮石斛 糖尿病 降血糖 腸道菌群

【摘要】：目的:研究人工培育的鐵皮石斛水提取物,對(duì)鏈脲霉素(STZ)所致的高血糖小鼠血糖、血脂、機(jī)體的脂肪和肌肉組織成分比例和肝臟抗氧化能力的影響,分析對(duì)腸道菌群影響。通過(guò)多種途徑分析高血糖對(duì)機(jī)體的影響,以及鐵皮石斛水提物對(duì)機(jī)體的保護(hù)作用和可能的機(jī)制。方法:腹腔注射STZ,制備小鼠糖尿病模型。實(shí)驗(yàn)分為鐵皮石斛水提物高、中、低劑量組、正常對(duì)照組、模型對(duì)照組、陽(yáng)性藥物組。觀察實(shí)驗(yàn)動(dòng)物的一般情況,記錄小鼠體重,用核磁共振儀測(cè)定小鼠機(jī)體組織成分比例,用血糖試紙測(cè)空腹血糖值,血清TC、TG、HDL值按照試劑盒說(shuō)明檢測(cè),利用分光光度計(jì)測(cè)肝臟SOD含量(黃嘌呤氧化酶法)、TBA法測(cè)MDA含量,通過(guò)PCR-DGGE凝膠電泳,quantity one軟件分析小鼠腸道菌群多樣性。結(jié)果:(1)正常對(duì)照組小鼠在整個(gè)實(shí)驗(yàn)過(guò)程中,生長(zhǎng)情況良好,沒(méi)有死亡現(xiàn)象。成模小鼠精神不振、反應(yīng)慢、不愛(ài)活動(dòng),每日喝水量增加,排尿次數(shù)明顯增加、體重明顯下降(P0.05)、毛發(fā)松馳無(wú)光澤、好激怒、糞便變小,變黃且發(fā)干,模型對(duì)照組存活狀態(tài)最差,存活率僅為50%。石斛水提物三個(gè)劑量組和陽(yáng)性藥物組小鼠的一般狀態(tài)均明顯改善,毛色和精神狀態(tài)有所恢復(fù),飲水量和尿量減少,糞便著色變深,存活率分別為80%、60%、60%、80%。模型對(duì)照組機(jī)體脂肪與肌肉的比例:正常對(duì)照組小鼠,隨體重的增加,脂肪組織明顯增加,明顯高于其他各組,脂肪組織所占比例肌肉組織。模型對(duì)照組小鼠,隨體重的下降,脂肪組織也減少,與其他各組相比有很大差別,脂肪組織所占比例肌肉組織。石斛水提物三個(gè)劑量組及陽(yáng)性藥物組,脂肪組織的變化與體重的變化一致,不論是脂肪組織的占比例還是肌肉組織所占比例均高于模型組,并且肌肉組織所占比例高于正常對(duì)照組。(2)與正常對(duì)照組比較,模型對(duì)照組血糖水平顯著上升,在建模第28d后開(kāi)始下降,3個(gè)石斛水提物組血糖水平在第7d起與模型對(duì)照組比有所減少,14~28d減少更加明顯,差異有顯著性(p0.01),這一結(jié)果與陽(yáng)性藥物組的血糖的變化一致,并且各組彼此沒(méi)有顯著差異(p0.05)。(3)建模28d后,與正常對(duì)照組比較,模型對(duì)照組小鼠血清tc、tg含量明顯高于正常對(duì)照組小鼠(p0.05),hdl低于正常對(duì)照組(p0.05)。與模型對(duì)照組比較,石斛水提物三個(gè)劑量組和陽(yáng)性藥物組的血清tc含量明顯減少(p0.05),hdl含量明顯增加(p0.05)。石斛低劑量組、陽(yáng)性藥物組tg含量低于模型對(duì)照組(p0.05),石斛中、高劑量組的血清tg含量高于模型對(duì)照組(p0.05)。(4)與正常對(duì)照組比較,模型對(duì)照組小鼠肝組織sod活性顯著降低(p0.05),mda水平顯著上升(p0.01)。與模型對(duì)照組比,三個(gè)劑量石斛水提物的肝組織sod活性均明顯升高、mda含量均減少(p0.05);陽(yáng)性藥物組sod活力有所增加(p0.05),但是mda含量高于模型對(duì)照組小鼠。(5)與正常對(duì)照組比較,陽(yáng)性藥物組和模型對(duì)照組糞便中菌群條帶模糊不清,細(xì)菌表達(dá)量較低。石斛水提物中、高劑量組菌群多樣性好且表達(dá)量較高。聚類分析結(jié)果表明,石斛水提物低、中劑量組為一簇,菌群相似度最高,石斛高劑量組、模型對(duì)照組,陽(yáng)性藥物組和正常對(duì)照組為一簇,菌群相似度高,其中陽(yáng)性藥物組與正常對(duì)照組菌群高度相似,石斛水提物治療組中以高劑量組的菌群與正常對(duì)照組最為相似。以上結(jié)果表明,鐵皮石斛水提物中、高劑量均能改善糖尿病小鼠的腸道菌群數(shù)量和多樣性,在提高菌群數(shù)量上比陽(yáng)性藥物作用效果好。結(jié)論:(1)石斛水提物能夠改善糖尿病小鼠機(jī)體的健康情況和體重急劇下降的并發(fā)癥;(2)石斛水提物對(duì)stz誘導(dǎo)的糖尿病小鼠有顯著的降血糖功效,和陽(yáng)性藥物(鹽酸二甲基雙胍)效果相差不多;(3)石斛水提物能調(diào)節(jié)stz糖尿病小鼠機(jī)體脂肪與肌肉組織的比例,提示其可能調(diào)節(jié)了機(jī)體脂代謝,從而影響血糖水平;(4)石斛水提物能夠調(diào)節(jié)糖尿病導(dǎo)致的tc、tg升高、hdl降低,改善糖尿病小鼠的血脂混亂;(5)石斛水提物能夠通過(guò)升高糖尿病小鼠肝組織sod活性、降低mda水平,來(lái)增強(qiáng)肝臟抗氧化能力;(5)石斛水提物能夠調(diào)節(jié)糖尿病所導(dǎo)致小鼠腸道菌群失調(diào)、菌群數(shù)量減少。綜上所述,鐵皮石斛水提物通過(guò)調(diào)節(jié)機(jī)體血脂、肝臟抗氧化能力和腸道菌群,改善了糖尿病小鼠的脂代謝,增強(qiáng)機(jī)體免疫能力,進(jìn)而間接調(diào)節(jié)了血糖水平的穩(wěn)定。所以,人工培育的鐵皮石斛可以作為治療糖尿病的輔助藥物,能夠降低血糖水平及改善糖尿病所導(dǎo)致的部分并發(fā)癥,為未來(lái)糖尿病的預(yù)防和治療提供新思路。
[Abstract]:Objective: To study the artificial cultivation of Dendrobium candidum water extract, on streptozotocin (STZ) hyperglycemia mice induced by glucose, lipid, effect of adipose and muscle composition of body and liver antioxidant ability, analysis of effect on the intestinal flora. Through a variety of ways to analysis the effects of high blood glucose on the body, and Dendrobium the water extract of protective effect on the body and the possible mechanism. Methods: intraperitoneal injection of STZ, the preparation of the diabetic mouse model. The experiment was divided into Dendrobium candidum water extract high, low dose group, normal control group, model control group, positive drug group. General observation of experimental animal, records of body weight of mice determination of body composition of mice, using nuclear magnetic resonance instrument, with blood glucose test strips, fasting blood glucose, serum TC, TG, HDL value detection in accordance with the kit, using the spectrophotometer to measure the content of liver SOD (xanthine oxidase method), TBA method To measure the content of MDA, PCR-DGGE by gel electrophoresis, analysis of mouse intestinal flora diversity of quantity one software. Results: (1) normal control mice during the whole experiment, growth is good, there is no death phenomenon. Model mice Jingshenbuzhen, slow response, not love activities, daily water quantity increased, a significant increase in the number of urination body weight, decreased significantly (P0.05), hair loose without luster, good anger, stool becomes small, yellow and dry, the model control group survived the worst state, the general state of survival rate is only 50%. from aqueous extracts of three dose group and positive drug group mice were significantly improved, the colour and the mental state recovery, water intake and urine volume decreased, fecal staining and survival rate were 80%, 60%, 60%, the proportion of 80%. in model control group, body fat and muscle of the mice in normal control group, with the increasing of body weight, fat tissue increased significantly, significantly higher than the other In each group, the proportion of adipose tissue, muscle tissue. The model control group, with the decrease of body weight, fat tissue also decreased compared with other groups, there is a great difference between the proportion of adipose tissue, muscle tissue. The water extract of Dendrobium three dose group and positive drug group, consistent changes in adipose tissue and body weight. Whether the proportion of adipose tissue or muscle tissue proportion is higher than the model group, and muscle tissue proportion is higher than the normal control group. (2) compared with the normal control group, model control group, blood glucose levels increased significantly in the model, 28d began to decline after 3 d. water extraction group blood glucose level in the 7d and the model control group decreased, 14~28d decreased more obviously, there was significant difference (P0.01) changes, the result was in line with the positive drug group the blood sugar level, and the groups had no significant difference each other (P0.05). (3) modeling 28 After D, compared with normal control group, model control group, serum TC, TG were higher than that of normal control mice (P0.05), HDL lower than the normal control group (P0.05). Compared with the model group, the serum TC content of water extract of Dendrobium three dose group and positive drug group decreased significantly (P0.05) and the content of HDL increased significantly (P0.05). From the low dose group, positive drug group TG was lower than that of model group (P0.05), Dendrobium, the content of serum TG in high dose group was higher than that of the model control group (P0.05). (4) compared with the normal control group, model control group of mice liver tissue SOD activity decreased significantly (P0.05), the level of MDA increased significantly (P0.01). Compared with model control group, three dose of water extract of Dendrobium liver SOD activity were significantly increased, MDA content decreased (P0.05); the positive drug group increased the activity of SOD (P0.05), but the content of MDA was higher than the model control group (5 mice. With normal) According to group, positive drug group and model control group of fecal bacteria with blurred, bacterial expression is relatively low. The water extract of Dendrobium in high dose group, the bacterial diversity and high expression level. The results of cluster analysis showed that Shi Hushui extract low, middle dose group as a cluster, flora similarity the highest Dendrobium high dose group, model control group, positive drug group and normal control group as a cluster of bacteria, high similarity, the positive drug group and normal control group of bacteria was highly similar to that of Dendrobium water extract in the treatment group with the flora of high dose group and normal control group. The above results most similar show that the Dendrobium candidum water extract and intestinal microflora in high dose could improve diabetic mice, increase in number of bacteria than the positive drug effect. Conclusion: (1) Shi Hushui extract can improve diabetic mice body health and weight urgently Play down the complications; (2) Shi Hushui extract had a significant hypoglycemic effect in STZ induced diabetic mice, and positive drug (two methyl guanidine hydrochloride) effect is not much; (3) Shi Hushui extract can regulate the STZ diabetic mice body fat and muscle tissue proportion, suggesting that it may regulate the body lipid metabolism, thus affecting the blood glucose level; (4) Shi Hushui extract can regulate the diabetes induced TC, TG increased, HDL decreased, improve blood lipid of diabetic mice confusion; (5) Shi Hushui extract can increase the activity of SOD in liver tissue of diabetic mice, decrease the level of MDA, to enhance the antioxidant capacity of the liver; (5) Shi Hushui extract can regulate diabetes caused by intestinal flora of mice, the number of bacteria decreased. In summary, the water extract of Dendrobium officinale by regulating blood lipid, antioxidant capacity of liver and intestinal microflora, improve lipid metabolism in diabetic mice Thanks, enhance the immune ability, and indirectly regulate the stability of blood glucose level. Therefore, the artificial cultivation of Dendrobium candidum can be used as auxiliary medicine for treating diabetes, can reduce the level of blood glucose and improve complications due to diabetes, for future diabetes prevention and treatment to provide new ideas.

【學(xué)位授予單位】：大連醫(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)】：R285.5

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9 王英;“工廠”里種出鐵皮石斛[N];蘇州日?qǐng)?bào);2008年

10 收集整理 軍野;鐵皮石斛的文化淵源[N];世界報(bào);2008年

相關(guān)博士學(xué)位論文 前10條

1 孫婧;鐵皮石斛提取物及其活性成分毛蘭素對(duì)人乳腺癌細(xì)胞抑制作用的研究[D];吉林大學(xué);2016年

2 梁楚燕;基于衰老雌鼠模型探討鐵皮石斛“補(bǔ)腎”功效的科學(xué)內(nèi)涵[D];廣州中醫(yī)藥大學(xué);2016年

3 徐程;鐵皮石斛種質(zhì)資源與組培工廠化生產(chǎn)研究[D];浙江大學(xué);2006年

4 包英華;鐵皮石斛種質(zhì)資源的鑒定與評(píng)價(jià)研究[D];廣州中醫(yī)藥大學(xué);2014年

5 嚴(yán)亮;中國(guó)傳統(tǒng)蘭科藥用植物鐵皮石斛基因組及其生物學(xué)特性研究[D];吉林大學(xué);2014年

6 張澤錦;鐵皮石斛的光合碳同化途徑及其對(duì)環(huán)境變化的生理響應(yīng)[D];中國(guó)農(nóng)業(yè)大學(xué);2014年

7 趙興兵;超微鐵皮石斛對(duì)脾虛便秘小鼠腸道微生物的影響研究[D];湖南中醫(yī)藥大學(xué);2014年

8 李標(biāo);聲波刺激對(duì)鐵皮石斛生長(zhǎng)代謝的影響及其相關(guān)基因表達(dá)和克隆研究[D];重慶大學(xué);2007年

9 滕建北;鐵皮石斛與美花石斛莖尖生長(zhǎng)機(jī)理及其多糖累積規(guī)律研究[D];成都中醫(yī)藥大學(xué);2009年

10 關(guān)杰敏;鐵皮石斛良種繁育與GAP種植關(guān)鍵技術(shù)研究[D];廣州中醫(yī)藥大學(xué);2014年

相關(guān)碩士學(xué)位論文 前10條

1 諸燕;鐵皮石斛種質(zhì)資源收集與評(píng)價(jià)[D];浙江農(nóng)林大學(xué);2010年

2 辛甜;組培鐵皮石斛的品質(zhì)評(píng)價(jià)研究[D];遼寧中醫(yī)藥大學(xué);2011年

3 戴燕萍;鐵皮石斛生產(chǎn)質(zhì)量管理規(guī)范研究[D];浙江農(nóng)林大學(xué);2012年

4 王俊潔;鐵皮石斛體細(xì)胞無(wú)性變異的生藥學(xué)初步研究[D];云南中醫(yī)學(xué)院;2012年

5 李東賓;冷脅迫下鐵皮石斛抗寒基因的克隆與表達(dá)研究[D];浙江農(nóng)林大學(xué);2013年

6 馮云;鐵皮石斛儲(chǔ)藏與加工特性研究[D];華南理工大學(xué);2015年

7 鄧星燕;鐵皮石斛紫外指紋圖譜研究[D];云南民族大學(xué);2015年

8 朱夢(mèng)麗;鐵皮石斛甘露糖結(jié)合凝集素的分子建模與對(duì)接研究，，及NAC基因組序列的克隆[D];西南交通大學(xué);2015年

9 陳爾;鐵皮石斛組培快繁技術(shù)的優(yōu)化[D];中南林業(yè)科技大學(xué);2015年

10 任潔;鐵皮石斛組培苗培養(yǎng)及叢生芽增殖體系的建立[D];南京師范大學(xué);2015年

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