【摘要】：目的:(1)探討有氧運(yùn)動和Ala-Gln對T2DM心肌纖維化的干預(yù)作用和交互作用。(2)從心肌組織中氧化應(yīng)激/NF-κB通路的變化,探討運(yùn)動訓(xùn)練和Ala-Gln干預(yù)T2DM心肌纖維化形成的生理機(jī)制。方法:選用雄性SD大鼠80只,隨機(jī)抽取10只大鼠作為正常對照組(NC,n=10),平時不運(yùn)動,標(biāo)準(zhǔn)飼料飼養(yǎng)。其余70只大鼠喂飼高脂膳食,6周后,在空腹?fàn)顟B(tài)下,注射小劑量的STZ(40mg/Kg體重),7天后,測試空腹血糖,當(dāng)血糖達(dá)到11.1mmol/L為建模成功。再將建模成功的T2DM大鼠隨機(jī)4組:糖尿病對照組(DMC,n=10)、糖尿病運(yùn)動組(DME,n=10)、糖尿病+Ala-Gln 組(DMAG,n=10)和糖尿病+Ala-Gln+運(yùn)動組(DMEAG,n=10),標(biāo)準(zhǔn)飼料飼養(yǎng)。DMC組平時不運(yùn)動;運(yùn)動鍛煉采用60min無負(fù)重的游泳運(yùn)動,每周6次;Ala-Gln在每次訓(xùn)練結(jié)束30min后灌服,補(bǔ)充劑量為1.5 g/Kg體重,每周6次。8周后,通過Elisa檢測各組大鼠血清中空腹血糖(FBG)、胰島素(INS)、C肽、胰高血糖素樣肽-1(GLP-1)、心肌鈣蛋白Ⅰ(cTnI)的含量和心肌組織核轉(zhuǎn)錄因子-κB(NF-κB)、轉(zhuǎn)化生長因子-β1(TGF-β1)、腫瘤壞死因子-α(TNF-α)含量;通過RT-PCR檢測心肌組織硫氧還蛋白還原酶(TR)、硫氧還蛋白(Trx)、Ⅰ、Ⅲ型膠原纖維mRNA的表達(dá);通過硫代巴比妥酸法檢測心肌組織丙二醛(MDA);通過分光光度計檢測還原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG);以及分別觀察心肌組織形態(tài)結(jié)構(gòu)和超微結(jié)構(gòu)。結(jié)果:(1)與NC組相比,DMC組FBG(P0.05)顯著升高,血清GLP-1(P0.01)、C肽(P0.05)含量顯著降低,INS含量有所增加,但無顯著性差異(P0.05)。通過雙因素方差分析,有氧運(yùn)動可以使T2DM大鼠FBG極顯著降低(P0.01),血清C肽含量極顯著升高(P0.01),血清INS、GLP-1含量有所升高,但無顯著性差異(P0.05);補(bǔ)充Ala-Gln可以使T2DM大鼠FBG(P0.05)含量顯著降低,血清INS(P0.05)、GLP-1(P0.05)、C肽(P0.01)含量顯著升高;有氧運(yùn)動聯(lián)合Ala-Gln對升高T2DM大鼠血清INS、C肽、GLP-1含量有顯著交互作用(P0.05),但對降低FBG含量無顯著交互作用(P0.05)。(2)與NC組相比,DMC組大鼠心肌組織Ⅰ型膠原纖維mRNA(P0.05)、Ⅲ型膠原纖維mRNA(P0.05)表達(dá)量顯著升高。通過雙因素方差分析,有氧運(yùn)動使T2DM大鼠心肌組織Ⅰ、Ⅲ型膠原纖維mRNA表達(dá)顯著降低(P0.05);補(bǔ)充Ala-Gln使心肌組織Ⅰ、Ⅲ型膠原纖維mRNA的表達(dá)顯著降低(P0.05);有氧運(yùn)動聯(lián)合Ala-Gln對降低心肌組織Ⅰ、Ⅲ型膠原纖維mRNA表達(dá)無顯著交互作用(P0.05)。與NC組相比,DMC組心肌細(xì)胞中肌原纖維排列紊亂,并有溶解現(xiàn)象,Z線扭曲;心肌細(xì)胞內(nèi)線粒體大小不等,嵴模糊,數(shù)量減少。DME和DMAG組心肌細(xì)胞中肌原纖維排列較為整齊,Z線排列較整齊。線粒體數(shù)量多,形態(tài)大致正常。DMEAG組心肌細(xì)胞內(nèi)肌原纖維排列整齊,肌絲結(jié)構(gòu)清晰,Z線清晰而排列整齊,線粒體數(shù)量顯著增多,嵴非常清楚。與此同時,與NC組相比,DMC組血清cTnI(P0.01)含量顯著升高。通過雙因素方差分析,有氧運(yùn)動可使血清cTnI含量極顯著降低(P0.01);補(bǔ)充Ala-Gln可以使血清cTnI(P0.01)含量顯著降低;有氧運(yùn)動聯(lián)合Ala-Gln對降低血清cTnI含量無顯著交互作用(P0.05)。(3)與NC組相比,DMC組心肌組織MDA(P0.05)、GSSG(P0.01)含量顯著升高,GSH含量降低,但無顯著性差異(P0.05),GSH/GSSG比值極顯著降低(P0.01),TR、Trx mRNA的表達(dá)量顯著降低(P0.05)。通過雙因素方差分析可知,有氧運(yùn)動使心肌組織MDA、GSSG含量顯著降低(P0.05),GSH含量、GSH/GSSG比值和TrxmRNA表達(dá)升高,但無顯著性差異(P0.05),而TR mRNA表達(dá)量顯著升高(P0.05);補(bǔ)充Ala-Gln使心肌組織MDA(P0.01)、GSSG(P0.05)含量顯著降低,GSH含量(P0.05)、GSH/GSSG比值(P0.01)顯著升高,TR、Trx mRNA表達(dá)顯著增加(P0.05)。有氧運(yùn)動聯(lián)合Ala-Gln對降低心肌組織MDA、GSSG含量和升高GSH含量、GSH/GSSG比值和TRmRNA表達(dá)均無顯著的交互作用(P0.05),但對升高Trx mRNA表達(dá)有顯著交互作用(P0.05)。(4)與NC組相比,DMC組心肌組織NF-κB、TGF-β1、TNF-α含量均顯著升高(P0.05)。通過雙因素方差分析,有氧運(yùn)動使心肌組織NF-κB、TGF-β1、TNF-α含量顯著降低(P0.01);補(bǔ)充 Ala-Gln 使 T2DM NF-κB(P0.01)、TGF-β1(P0.01)、TNF-α(P0.05)含量顯著降低;有氧運(yùn)動聯(lián)合Ala-Gln對降低心肌組織NF-κB含量無顯著交互作用(P0.05),但對降低TGF-β1(P0.01)和TNF-α(P0.05)含量有顯著的交互作用。結(jié)論:(1)本研究所復(fù)制的T2DM大鼠模型是成功的,單純有氧運(yùn)動、Ala-Gln可以促進(jìn)T2DM大鼠胰島素的分泌,對控制T2DM大鼠的FBG具有顯著的作用,而有氧運(yùn)動聯(lián)合Ala-Gln對降低T2DM大鼠的血糖更加有效。(2)T2DM大鼠在建模成功8周后,心肌組織就發(fā)生一定程度的纖維化,并引起心肌細(xì)胞發(fā)生一定程度的損傷。而有氧運(yùn)動和補(bǔ)充Ala-Gln可以有效地抑制T2DM大鼠心肌纖維化的形成,且有氧運(yùn)動聯(lián)合Ala-Gln更加有效。(3)T2DM大鼠心肌組織中硫氧還蛋白系統(tǒng)功能的降低,引起心肌氧化應(yīng)激,促進(jìn)心肌組織中炎性因子的分泌,是T2DM心肌纖維化形成的重要機(jī)制;而有氧運(yùn)動和補(bǔ)充Ala-Gln可提高T2DM心肌組織中硫氧還蛋白系統(tǒng)的功能,減少M(fèi)DA的生成,從而減輕心肌組織中炎性因子的分泌,抑制T2DM心肌纖維化的發(fā)生和發(fā)展。
[Abstract]:Objective: (1) To study the effect and interaction of aerobic exercise and Ala-Gln on myocardial fibrosis in patients with T2DM. (2) The physiological mechanism of the formation of myocardial fibrosis in T2DM with exercise training and Ala-Gln was discussed. Methods:80 male SD rats were randomly selected as the normal control group (NC, n = 10). The remaining 70 rats were fed with a high-fat diet and, after 6 weeks, a small dose of STZ (40 mg/ kg body weight) was injected, and after 7 days, the fasting blood glucose was tested and the model was successful when the blood glucose reached 11.1 mmol/ L. The model was divided into four groups: the diabetic control group (DMC, n = 10), the diabetic group (DME, n = 10), the diabetes + Ala-Gln group (DMAG, n = 10) and the diabetes + Ala-Gln + exercise group (DMAGAG, n = 10), and the standard feed. The dmc group did not move normally; the exercise exercise had no weight-bearing swimming exercise for 60 minutes,6 times a week; the Ala-Gln was taken after 30 minutes at the end of each training, and the supplementary dose was 1.5 g/ Kg body weight,6 times a week. After 8 weeks, fasting blood glucose (FBG), insulin (INS), and C-peptide in the serum of each group were detected by Elisa. The content of Glucagon-like peptide-1 (GLP-1), the content of cardiac troponin I (cTnI) and the nuclear transcription factor-B (NF-EMAB), the transformation growth factor-1 (TGF-1), and the tumor necrosis factor-1 (TNF-1) content; The expression of thioredoxin reductase (TR), thioredoxin (Trx), 鈪，

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