發(fā)布時(shí)間：2018-04-21 04:33

  本文選題：生酮飲食 + 運(yùn)動(dòng)　； 參考：《華東師范大學(xué)》2017年碩士論文

【摘要】：糖尿病是威脅全球人類身體健康的非傳染性疾病之一,不但給患者個(gè)人造成了極大的健康負(fù)擔(dān),而且也給國(guó)家和社會(huì)造成了沉重的經(jīng)濟(jì)負(fù)擔(dān)。國(guó)際糖尿病聯(lián)盟(IDF)最新統(tǒng)計(jì)數(shù)據(jù)顯示,2016全球約約有4.15億成年糖尿病患者,據(jù)估計(jì),2040年全球成年糖尿病患者將達(dá)到6.42億。糖尿病發(fā)病機(jī)制極為復(fù)雜,至今為止,尚無任何方法可完全治愈糖尿病。目前二型糖尿病主要治療手段有藥物治療、飲食治療以及合理的體育活動(dòng)。由于藥物的安全性以及除二甲雙胍外大多數(shù)糖尿病藥物療效有限或價(jià)格昂貴,因此,非藥療法越來越受到人們的重視。臨床研究發(fā)現(xiàn),生酮飲食(Ketogenic Diet,KD)作為一種糖尿病的干預(yù)手段可對(duì)患者的糖代謝穩(wěn)態(tài)產(chǎn)生良性影響,并開始用于二型糖尿病患者的臨床治療。然而,近幾年有研究發(fā)現(xiàn)糖尿病小鼠長(zhǎng)期生酮飲食可能導(dǎo)致肝臟組織脂肪代謝紊亂,甚至誘發(fā)脂肪肝。臨床及動(dòng)物研究顯示不同方式運(yùn)動(dòng)對(duì)脂肪肝的預(yù)防、治療均有一定的積極作用,但是運(yùn)動(dòng)能否改善生酮飲食治療所導(dǎo)致的肝臟組織脂肪代謝紊亂仍待研究。目的:(1)探究生酮飲食對(duì)糖尿病小鼠糖、脂代謝的影響及其機(jī)制。(2)探究生酮飲食聯(lián)合不同方式運(yùn)動(dòng)對(duì)糖尿小鼠肝臟脂代謝影響及其機(jī)制。方法:雄性C57BL/6J小鼠(3-5周齡),共85只,適應(yīng)性飼養(yǎng)3天,隨機(jī)挑選出10只作為正常對(duì)照組(NC),采用標(biāo)準(zhǔn)普通飼料喂養(yǎng)。另外75只小鼠首先采用45%高脂飼料喂養(yǎng)4周,之后100 mg/kg BW腹腔注射STZ,連續(xù)三次空腹血糖水平高于11.1 mmol/l,判斷為Ⅱ型糖尿病造模成功。此次實(shí)驗(yàn)中共有58只小鼠造模成功,根據(jù)體重和血糖水平選取32只,將其分為糖尿病對(duì)照組(HFD,n=8)、生酮飲食組(KD,n=8)、生酮飲食高強(qiáng)度間歇運(yùn)動(dòng)組(KH,n=8)、生酮飲食有氧運(yùn)動(dòng)組(KA,n=8)。每次運(yùn)動(dòng)前、后進(jìn)行5min,8m/min的準(zhǔn)備活動(dòng)和整理活動(dòng),KH組采取8周無坡度跑臺(tái)訓(xùn)練,高強(qiáng)度運(yùn)動(dòng)4 min,積極性休息2 min,速度由17m/min增至24m/min,每次運(yùn)動(dòng)60min,3次/周;KA組采取8周無坡度跑臺(tái)訓(xùn)練,12m/min速度運(yùn)動(dòng)60min,每次運(yùn)動(dòng)60min,5次/周。結(jié)果:(1)與 NC 組相比較,HFD 組小鼠 FBG(p0.01)、HbAlc(p0.05),血清 Glu(p0.05)、Insulin(p0.05),HOMA-IR(p0.05)及日飲水量(p0.05)顯著上升,HOMA-ISI(p0.05)顯著降低;KD組HOMA-ISI(p0.05)顯著上升,血清 Insulin(p0.05)、HOMA-IR(p0.05)體重(p0.05)顯著下降,FBG、HbAlc、血清Glu以及日飲水量無顯著差異;KH組HOMA-ISI(p0.05)顯著上升,HOMA-IR(p0.05)、血清 Insulin(p0.05)和體重(p0.05)顯著下降,FBG、HbAlc、血清Glu及日飲水量無顯著差異;KA組小鼠血清Insulin(p0.05)、體重(p0.05)顯著下降,FBG、HbAlc、血清 Glu、HOMA-IR、HOMA-ISI 以及日飲水量無顯著差異;與HFD組相比較,KD組、KH組以及KA組HOMA-ISI(p0.05)、均顯著升高;KD 組、KH 組以及 KA 組 FBG(p0.01)、HbAlc(p0.05),血清 Insulin(p0.05)、Glu(p0.05),HOMA-IR(p0.05)、體重(p0.05)及日飲水量(p0.05)顯著下降。與NC組相比較,KD組、KH組以及KA組小鼠G6PC以及PCK1 mRNA表達(dá)均顯著降低(p0.01);與HFD組相比較,KD組、KH組、KA組小鼠G6PC以及PCK1 mRNA表達(dá)均顯著降低(p0.01)。(2)與 NC 組相比較,HFD 組血清 ALT(p0.01)、TC(p0.05)、LDL-C(p0.05)、肝指數(shù)(p0.01)、肝臟TG(p0.01)、NEFA(p0.01)水平顯著升高,附睪脂肪%顯著降低(p0.05),血清TG、HDL-C、AST水平無顯著變化;KD組血清 TC(p0.05)、TG(p0.05)、HDL-C(p0.05)、ALT(p0.01)、AST(p0.05),附睪脂肪%(p0.01)、肝指數(shù)(p0.01)、肝臟 TG(p0.01)、NEFA(p0.05)顯著升高;與HFD組相比較,KD組血清HDL-C(p0.05)、附睪脂肪%(p0.01)顯著升高,血清LDL-C水平、肝指數(shù)顯著降低(p0.05),血清AST、ALT、TG、TC及肝臟組織TG、NEFA水平無顯著差異。肝臟HE以及油紅染色顯示,NC組小鼠肝臟的肝小葉結(jié)構(gòu)完整,HFD組小鼠肝臟脂肪空泡較多,細(xì)胞膜結(jié)構(gòu)不清晰,KD組小鼠肝臟也出現(xiàn)較多明顯的脂肪空泡;肝臟MASSON染色顯示,與NC組相比較,HFD組及KD組小鼠肝臟組織均出現(xiàn)了不同程度的纖維化。(3)與NC組相比,HFD組小鼠肝臟組織脂代謝相關(guān)基因CIDEA(p0.01)、SCD1(p0.05)、ACC1(p0.01)、ACC2(p0.01)、SREBP1(p0.05)、CPT1A(p0.001)的mRNA水平出現(xiàn)顯著上升,FAS、AMPKa1的mRNA表達(dá)水平未有顯著變化,AMPKa2的mRNA水平顯著降低(p0.05);KD組CIDEA、FAS、SCD1、SREBP1、ACC1、ACC2、CPT1A 的 mRNA 水平均顯著上升(p0.01),AMPKa1(p0.05)、AMPKa2(p0.01)的 mRNA 表達(dá)水平顯著下降;與 HFD組相比較可以發(fā)現(xiàn),生KD組小鼠肝臟組織AMPKa2的mRNA表達(dá)水平顯著下降,SCD1(p0.05)、AMPKa2(p0.05)以及 CPT1A(p0.01)的 mRNA 相對(duì)表達(dá)均顯著上升,CIDEA、FAS、SREBP1、ACC1、ACC2、AMPKa1 的 mRNA表達(dá)水平未有顯著差異。(4)與NC組相比,HFD組小鼠肝臟的FAS蛋白表達(dá)水平顯著上升(p0.01),ATGL、P-ACC/ACC 蛋白表達(dá)水平出現(xiàn)顯著下降(p0.05),HSL、MGL、PPARα、PPARγ、CPT1A蛋白表達(dá)水平以及P-AMPK/AMPK蛋白比值未發(fā)生顯著變化,生酮飲食組小鼠肝臟P-AMPK/AMPK蛋白比值出現(xiàn)顯著下降(p0.05),FAS(p0.01)、PPARγ(p0.05)、CPT1A(p0.05)蛋白表達(dá)出現(xiàn)顯著上升,HSL、ATGL、MPGL、PPARα蛋白表達(dá)以及P-ACC/ACC蛋白比值無顯著變化;與糖尿病對(duì)照組相比,生酮飲食組小鼠肝臟FAS蛋白及P-AMPK/AMPK蛋白表達(dá)比值顯著下降(p0.05),ATGL、CPT1A、PPARα蛋白表達(dá)顯著提高(p0.05),而HLS、MGL、PPARy蛋白表達(dá)以及P-ACC/ACC蛋白比值無顯著變化。(5)與KD組相比較,KA組糖尿病小鼠血清TG(p0.05)、ALT(p0.01)、AST(p0.05)、HDL-C(p0.01),肝臟 TG(p0.01)、NEFA(p0.01)水平顯著降低,血清LDL-C(p0.01)的水平顯著升高;肝指數(shù)及附睪脂肪%未發(fā)生明顯差異,KH 組血清 LDL-C(p0.01)水平顯著提高,血清 ALT(p0.05)、TC(p0.05)、肝臟TG(p0.05)水平顯著降低,血清TG、AST水平、肝臟組織NEFA含量及肝指數(shù)、附睪脂肪%未發(fā)生明顯變化。肝臟HE、油紅染色顯示,與KD組相比,KA組小鼠肝細(xì)胞形態(tài)趨向于正常,脂肪脂滴含量明顯減少,KH組肝細(xì)胞結(jié)構(gòu)較為清晰,肝細(xì)胞內(nèi)脂滴含量減少;肝臟MASSON染色顯示,與KD組相比,KH纖維化程度稍有降低,KA組肝臟纖維化顯著降低。(6)與KD組相比,KH組及KA組小鼠肝臟組織G6PC的mRNA相對(duì)表達(dá)量均顯著降低(p0.05),KA組小鼠肝臟組織G6PC活性顯著下降(p0.05),而KH組無顯著差異。與KD組相比,生酮飲食聯(lián)合高強(qiáng)度間歇運(yùn)動(dòng)或有氧運(yùn)動(dòng)干預(yù)均可使糖尿病小鼠肝臟PCK1的mRNA相對(duì)表達(dá)量顯著降低(p0.01),且均可顯著降低小鼠肝臟組織PCK1的活性(p0.05)。與KD組相比,KH組小鼠肝臟組織 CIDEA(p0.01)、FAS(p0.01)、SCD1(p0.05)、SREBP1(p0.01)、ACC1(p0.01)的 mRNA 相對(duì)表達(dá)顯著降低,ACC2(p0.05)、AMPKa2(p0.01)、CPT1A(p0.05)的mRNA相對(duì)表達(dá)含量顯著提高,AMPKa1的mRNA影響則不明顯。KA組小鼠肝臟組織CIDEA、FAS、SCD1、SREBP1、ACC1、ACC2的mRNA相對(duì)表達(dá)量(p0.01)顯著降低,肝臟AMPKa1、AMPKa2、CPT1A(p0.01)的mRNA相對(duì)表達(dá)量顯著提高。(7)與KD組相比較,KA組肝臟FAS、HSL、ATGL蛋白表達(dá)無顯著變化,MGL、CPT1A的蛋白表達(dá)、P-AMPK/AMPK、P-ACC/ACC蛋白表達(dá)的比值均顯著升高(p0.05),KH 組 HSL、ATGL、MGL、CPT1A 的蛋白表達(dá)、P-ACC/ACC蛋白表達(dá)的比值均無顯著變化,FAS蛋白表達(dá)顯著降低(p0.05)P-AMPK/AMPK蛋白表達(dá)的比值均顯著上升(p0.05)。結(jié)論:(1)生酮飲食及生酮飲食聯(lián)合運(yùn)動(dòng)干預(yù)均可顯著改善STZ誘導(dǎo)的Ⅱ型糖尿病小鼠血糖控制。(2)長(zhǎng)期生酮飲食,有可能會(huì)導(dǎo)致STZ誘導(dǎo)的T2DM小鼠肝臟組織脂肪積累,產(chǎn)生脂肪肝。(3)生酮飲食結(jié)合有氧運(yùn)動(dòng)干預(yù)或生酮飲食結(jié)合高強(qiáng)度間歇運(yùn)動(dòng)干預(yù),均可有效緩解單獨(dú)使用生酮飲食所導(dǎo)致糖尿病小鼠肝臟組織脂肪積累,且有氧運(yùn)動(dòng)效果更明顯。(4)有氧運(yùn)動(dòng)可能通過激活A(yù)MPK/ACC/CPT1A信號(hào)通路加速糖尿病小鼠肝臟組織脂肪氧化,減緩脂肪肝發(fā)生進(jìn)程,而高強(qiáng)度間歇運(yùn)動(dòng)則不依賴此調(diào)節(jié)途徑。
[Abstract]:Diabetes is one of the non communicable diseases that threaten the health of the world. It not only creates a huge health burden on individuals, but also creates a heavy economic burden on the country and society. The latest statistics of the International Diabetes Association (IDF) show that about 415 million adult diabetic patients around the world are about 2016, estimated to be in 2040. The incidence of diabetes in adults will reach 642 million. The pathogenesis of diabetes is very complicated. So far, there is no way to cure diabetes completely. At present, the main treatment of type two diabetes is medications, diet and physical exercise. Nondrug therapy is becoming more and more important. Therefore, non drug therapy is becoming more and more important. Clinical studies have found that Ketogenic Diet (KD), as an intervention for diabetes, can have a benign effect on the homeostasis of glucose metabolism in patients and began to be used in the clinical treatment of patients with type two glycometabolism. However, in recent years, there have been studies on the discovery of sugar. The long-term ketone diet may lead to lipid metabolism disorder in the liver tissue and even induce fatty liver. Clinical and animal studies have shown that different ways of exercise have positive effects on the prevention and treatment of fatty liver. But whether the exercise can improve the lipid metabolism disorder in the liver tissue caused by the treatment of ketogenic diet still needs to be studied. (1) to explore the effect of ketogenic diet on glucose and lipid metabolism in diabetic mice and its mechanism. (2) to explore the effect and mechanism of ketogenic diet combined with different exercise on liver lipid metabolism in diabetic mice. Methods: male C57BL/6J mice (3-5 weeks of age), 85, adaptive feeding for 3 days, and randomly selected 10 as normal control group (NC), using standard general popularization The other 75 mice were fed with 45% high fat diet for 4 weeks, and then 100 mg/kg BW were intraperitoneally injected with STZ, and three consecutive fasting blood glucose levels were higher than 11.1 mmol/l. It was judged to be a successful model of type II diabetes. In this experiment, 58 mice were successfully built, and 32 were selected according to the weight and blood glucose level, and they were divided into diabetes pairs. The group (HFD, n=8), the ketogenic diet group (KD, n=8), the high intensity intermittent exercise group (KH, n=8) and the aerobic exercise group (KA, n=8) in the ketogenic diet (KA, n=8). Before each exercise, the preparation activities and the activities of 8m/min were carried out. The KH group took 8 weeks without slope running platform training, the high intensity exercise was 4 min, and the active rest was 2. The speed was increased from 2 Each exercise was 60min, 3 times per week, group KA took 8 weeks without slope running, and 12m/min speed exercise was 60min, each exercise was 60min, 5 times per week. (1) compared with group NC, HFD group mice FBG (P0.01), HbAlc (P0.05), serum Glu and daily drinking water increased significantly. HOMA-ISI (P0.05) increased significantly, serum Insulin (P0.05), HOMA-IR (P0.05) weight (P0.05) decreased significantly, FBG, HbAlc, serum Glu and daily drinking water had no significant difference. Serum Insulin (P0.05), body weight (P0.05) decreased significantly, FBG, HbAlc, serum Glu, HOMA-IR, HOMA-ISI and daily drinking water amount had no significant difference. Compared with the HFD group, KD, KH and KA groups were all significantly higher. Weight (P0.05) and daily drinking water (P0.05) decreased significantly. Compared with the NC group, the G6PC and PCK1 mRNA expressions in KD, KH and KA groups were significantly decreased (P0.01). The liver index (P0.01), liver TG (P0.01), NEFA (P0.01) level significantly increased, the epididymal fat% decreased significantly (P0.05), the serum TG, HDL-C, AST level had no significant changes; KD group serum TC (P0.05), fat% of epididymis, liver index, liver index, significantly higher; The serum HDL-C (P0.05) and epididymal fat% (P0.01) in the KD group were significantly higher, the serum LDL-C level, the liver index decreased significantly (P0.05), the serum AST, ALT, TG, TC and liver tissue TG were not significant differences. The liver MASSON staining showed that the liver tissues of group HFD and KD group had different degrees of fibrosis in KD group. (3) compared with the NC group, the lipid metabolism related gene CIDEA (P0.01), SCD1 (P0.05), ACC1 (P0.05) in the HFD group were compared with the NC group. 5, the level of mRNA in CPT1A (p0.001) increased significantly, and the level of mRNA expression in FAS and AMPKa1 was not significantly changed, and the mRNA level of AMPKa2 decreased significantly (P0.05). The expression level of mRNA in the liver tissue of KD mice decreased significantly, and the relative expressions of SCD1 (P0.05), AMPKa2 (P0.05) and CPT1A (P0.01) were significantly increased. (4) there was no significant difference in the expression level of the mRNA. The expression level of P0.01, ATGL and P-ACC/ACC decreased significantly (P0.05), HSL, MGL, PPAR a, PPAR gamma, CPT1A protein expression level and P-AMPK/AMPK protein ratio did not change significantly, the ratio of P-AMPK/AMPK protein in the liver of the ketone diet mice decreased significantly (P0.05). There was no significant increase in the expression of HSL, ATGL, MPGL, PPAR alpha and P-ACC/ACC protein. Compared with the control group, the ratio of FAS protein and P-AMPK/AMPK protein in the liver of the ketone diet group decreased significantly (P0.05), ATGL, CPT1A, and PPAR alpha protein expression significantly increased (P0.05). There was no significant change in white ratio. (5) compared with group KD, serum TG (P0.05), ALT (P0.01), AST (P0.05), HDL-C (P0.01), liver TG (P0.01), liver TG (P0.01), liver index and epididymal fat% were significantly higher in KA group. ALT (P0.05), TC (P0.05), liver TG (P0.05) level decreased significantly, serum TG, AST level, liver tissue NEFA content and liver index, epididymal fat% did not change obviously. Liver HE, oil red staining showed that the hepatocyte morphology of KA group mice tended to be normal, fat lipid droplet content decreased obviously, liver cell structure was clearer, The lipid droplet content in liver cells decreased, and the liver MASSON staining showed that the degree of KH fibrosis decreased slightly and the liver fibrosis in the group KA decreased significantly compared with the KD group. (6) the mRNA relative expression of G6PC in the KH and KA groups decreased significantly (P0.05), and the G6PC activity of the liver tissue in the KA group was significantly lower than that in the KD group. Compared with the KD group, the relative expression of PCK1 mRNA in the liver of diabetic mice could be reduced significantly (P0.01) by combination of ketogenic diet combined with high intensity intermittent exercise or aerobic exercise (P0.01), and the activity of PCK1 in liver tissue of mice was significantly reduced (P0.05). The liver tissue of KH mice was compared with that of the KD group. The relative expression of mRNA in BP1 (P0.01) and ACC1 (P0.01) decreased significantly, while ACC2 (P0.05), AMPKa2 (P0.01), CPT1A (P0.05) increased significantly. (7) there was no significant change in the expression of FAS, HSL, ATGL protein in the liver of KA group compared with the KD group. The protein expression of MGL, CPT1A, P-AMPK/AMPK, and P-ACC/ACC protein expression increased significantly (P0.05). The ratio of the expression of (P0.05) P-AMPK/AMPK protein increased significantly (P0.05). Conclusion: (1) the combination of ketogenic diet and ketogenic diet combined exercise could significantly improve the control of blood glucose in type II diabetic mice induced by STZ. (2) the long-term ketone diet may lead to the accumulation of fat in the liver tissue of STZ induced T2DM mice and the production of fatty liver. (3) Ketone diet combined with aerobic exercise intervention or ketogenic diet combined with high intensity intermittent exercise intervention can effectively alleviate the lipid accumulation in liver tissue of diabetic mice induced by single use of ketone diet, and the effect of aerobic exercise is more obvious. (4) aerobic exercise may accelerate the liver tissue lipid in diabetic mice by activating the AMPK/ACC/ CPT1A signaling pathway Fat oxidation slowed down the development of fatty liver, while high intensity intermittent exercise did not depend on this regulation.

【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R587.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 朱兵;張曉雨;范鳴;杜冰;;高脂低碳水化合物飼養(yǎng)加運(yùn)動(dòng)對(duì)2型糖尿病大鼠代謝指標(biāo)的影響[J];中華糖尿病雜志;2014年11期

，

本文編號(hào)：1780963