【摘要】：目的:通過觀察替米沙坦(telmisartan)對(duì)1型糖尿病(T1DM)大鼠睪丸脂聯(lián)素(Adiponectin,APN)、脂聯(lián)素受體(Adipo R)和腺苷酸活化蛋白激酶(AMPK)和蛋白激酶B/內(nèi)皮性一氧化氮合酶/一氧化氮(Akt/e NOS/NO)表達(dá)的影響,探討替米沙坦如何保護(hù)T1DM大鼠睪丸。方法:雄性SD大鼠27只隨機(jī)分為正常對(duì)照組NC組8只和糖尿病造模組19只。用鏈脲佐菌素(STZ)誘導(dǎo)T1DM模型,16只成功。造模成功的大鼠被隨機(jī)分為糖尿病組DM組8只和替米沙坦治療組DT組8只。給DT組替米沙坦灌胃,NC組和DM組等量生理鹽水灌胃。替米沙坦治療8周后留取血標(biāo)本,用于檢測(cè)相關(guān)指標(biāo),然后處死動(dòng)物并取出雙側(cè)睪丸,同時(shí)剪下附睪制備精子懸液。用40%甲醛溶液固定部分睪丸,HE染色;其余睪丸經(jīng)液氮冷凍透后于-70℃冰箱待用。放免法測(cè)胰島素(Insulin)及性激素。酶聯(lián)免疫吸附(ELISA)法測(cè)脂聯(lián)素(APN)、白介素6(IL-6)和腫瘤壞死因子α(TNF-α)。用實(shí)時(shí)熒光定量PCR測(cè)Adipo R m RNA的表達(dá)。脂聯(lián)素及其受體、AMPK、P-AMPK、AKT、P-AKT和e-NOS蛋白表達(dá)用免疫印跡法(Western Blot)測(cè)。結(jié)果:1.各組大鼠的體重、睪丸指數(shù)、精子數(shù)目、精子活動(dòng)率的比較DM組較NC組大鼠睪丸重量明顯減輕(P0.05),睪丸指數(shù)無明顯變化(P0.05),精子數(shù)目、精子活動(dòng)率明顯降低(P0.05)。8周telmisartan治療后,DT組大鼠睪丸重量、睪丸指數(shù)無明顯變化(P0.05),精子數(shù)目、精子活動(dòng)率明顯升高(P0.05)。2.各組大鼠血糖、TCH、TG、FFA、性激素、Insulin、APN、睪丸NO、IL-6、TNF-α表達(dá)的比較與NC組比較,DM組大鼠血糖、TCH、TG、FFA、睪丸組織NO、IL-6、TNF-α均明顯升高(P0.05),血清及睪丸組織睪酮(T)、血清及睪丸組織脂聯(lián)素(APN)、胰島素(Insulin)顯著降低(P0.05),LH和FSH差異無統(tǒng)計(jì)學(xué)意義(P0.05)。8周替米沙坦治療后,DT組大鼠血糖、TCH、TG、FFA、NO、IL-6、TNF-α均明顯降低(P0.05),血清及睪丸組織APN明顯上升(P0.05),血清及睪丸組織T、Insulin、LH、FSH無明顯變化(P0.05)。3.各組大鼠睪丸組織形態(tài)學(xué)觀察比較NC組睪丸組織生精小管及細(xì)胞排列正常,管腔內(nèi)精子較多。DM組睪丸組織曲細(xì)精管萎縮變形,管腔內(nèi)空泡狀變性明顯,生精細(xì)胞數(shù)量明顯減少,生精小管之間的間質(zhì)組織比例增加,但間質(zhì)細(xì)胞體積縮小,胞漿減少,核固縮,間質(zhì)組織中微血管腔閉塞、淤血,炎性細(xì)胞浸潤(rùn)于睪丸間質(zhì)。DT組睪丸組織變性較DM組明顯好轉(zhuǎn)。4.各組大鼠睪丸脂聯(lián)素受體1、2 m RNA的表達(dá)水平比較與NC組比較,DM組大鼠睪脂聯(lián)素受體1 m RNA表達(dá)降低[(0.54±0.14)對(duì)(0.71±0.15),P0.05],脂聯(lián)素受體2 m RNA表達(dá)無明顯變化[(0.67±0.11)對(duì)(0.77±0.16),P0.05]。與DM組比較,DT組大鼠脂聯(lián)素受體1和2 m RNA表達(dá)均無明顯變化[(0.61±0.07)對(duì)(0.54±0.14)、(0.55±0.09)對(duì)(0.67±0.11),P0.05]。5.睪丸脂聯(lián)素及其受體、P-AMPK/AMPK、P-AKT/Akt、e NOS蛋白表達(dá)水平的比較DM組較NC組大鼠睪丸脂聯(lián)素、脂聯(lián)素受體1、P-AMPK/AMPK蛋白表達(dá)均明顯降低[(0.66±0.09)對(duì)(1.00±0.00),(0.68±0.05)對(duì)(1.00±0.00),(0.34±0.11)對(duì)(1.00±0.00),均P0.05],P-AKT/AKT和e NOS蛋白表達(dá)顯著增高[(1.54±0.27)對(duì)(1.00±0.00),(1.56±0.26)對(duì)(1.00±0.00),P0.05],Adipo R2蛋白表達(dá)無明顯變化[(1.02±0.13)對(duì)(1.00±0.00),P0.05]。替米沙坦治療后,DT組大鼠睪丸脂聯(lián)素和脂聯(lián)素受體1蛋白表達(dá)顯著增高[(0.84±0.09)對(duì)(0.66±0.09),(0.80±0.07)對(duì)(0.68±0.05),P0.05],P-AKT/AKT和e NOS表達(dá)蛋白顯著降低[(1.24±0.39)對(duì)(1.54±0.27),(1.16±0.47)對(duì)(1.56±0.26),P0.05],而P-AMPK/AMPK和脂聯(lián)素受體2蛋白表達(dá)無明顯變化[(0.65±0.52)對(duì)(0.34±0.11),(1.02±0.15)對(duì)(1.02±0.13),P0.05]。結(jié)論:替米沙坦通過對(duì)APN、Adipo R的調(diào)節(jié)以及Adipo R介導(dǎo)的信號(hào)通路AKT/e NOS/NO的影響減輕糖尿病大鼠睪丸組織損傷程度,對(duì)T1DM大鼠睪丸組織產(chǎn)生保護(hù)的作用。
[Abstract]:AIM: To investigate the effects of telmisartan on the expression of adiponectin (APN), adiponectin receptor (Adipo R), adenylate-activated protein kinase (AMPK) and protein kinase B/endothelial nitric oxide synthase/nitric oxide (Akt/e NOS/NO) in testis of type 1 diabetic (T1DM) rats, and to explore how telmisartan protects testis of T1DM rats. Methods: Twenty-seven male SD rats were randomly divided into normal control group (NC group, 8 rats) and diabetic model group (DM group, 19 rats). T1DM model was induced by streptozotocin (STZ), and 16 rats were successfully established. After 8 weeks of telmisartan treatment, blood specimens were taken for testing the related indexes. Then the animals were sacrificed and the testicles were taken out. At the same time, the epididymis was cut off to prepare sperm suspension. Adiponectin (APN), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by adsorption (ELISA). The expression of Adipo R m RNA was measured by real-time fluorescence quantitative PCR. The expressions of adiponectin and its receptors, AMPK, P-AMPK, AKT, P-AKT and e-NOS were detected by Western Blot. Results: 1. Body weight, testicular index, sperm number, sperm viability, and sperm motility were measured by Western Blot. Compared with NC group, the testicular weight and testicular index of DM group decreased significantly (P 0.05), testicular index did not change significantly (P 0.05), sperm number and sperm motility decreased significantly (P 0.05). Compared with NC group, the expression of glucose, TCH, TG, FFA, sex hormone, Insulin, APN, NO, IL-6 and TNF-alpha in DM group were significantly increased (P 0.05), serum and testicular tissue testosterone (T), serum and testicular adiponectin (APN), insulin (Insulin) were significantly decreased (P 0.05), LH and FSH were not significantly different between DM group and NC group. After 8 weeks of telmisartan treatment, blood glucose, TCH, TG, FFA, NO, IL-6, TNF-alpha were significantly decreased in DT group (P 0.05), APN in serum and testicular tissue were significantly increased (P 0.05), T, Insulin, LH, FSH in serum and testicular tissue were not significantly changed (P 0.05). 3. Morphological observation of testicular tissue in each group was compared with NC group. In DM group, the seminiferous tubules were atrophied and deformed, the number of spermatogenic cells was significantly reduced, the proportion of interstitial tissue between seminiferous tubules was increased, but the volume of interstitial cells was reduced, cytoplasm was reduced, nuclear pyknosis, microvascular lumen occlusion, congestion and inflammatory cells infiltrated into the testis. Compared with NC group, the expression of adiponectin receptor 1m RNA in DM group decreased [(0.54 +0.14) to ((0.71 +0.15), P 0.05], and the expression of adiponectin receptor 2m RNA did not change significantly [(0.67 +0.11) to ((0.77 +0.16), P 0.05]. There was no significant change in adiponectin receptor 1 and 2 m RNA expression in DT group [(0.61 +0.07) vs (0.54 +0.14), (0.55 +0.09) vs (0.67 +0.11), P.05]. 5. Expression of adiponectin and its receptors, P-AMPK/AMPK, P-AKT/Akt, e NOS protein in DM group compared with NC group. P-AKT/AKT/AKT and E NOS protein expression significantly increased [(1.54 [(1.54 [(0.27) P-AKT/AKT/AKT/AKT and E NOS protein expression significantly increased [(1.54 [(1.54 [(1.54 [(1.54 [(1.54 [(1.54 [(0.27) 1.00 [(1.00 [(1.56 [(1.56 [(1.56 0.26) 1.56 [(1.56 [(1.00 [0.56 [0.56 0.26)] (1.00 [(0.00 [0.00 [0.00 0.00 [0.0.00, P 0.05].Telmisartan After treatment, the expressions of adiponectin and adiponectin receptor 1 in the testis of DT rats were significantly increased [(0.84 65507 CONCLUSION: Telmisartan attenuates testicular tissue injury in diabetic rats by regulating APN, Adipo R and the signal pathway AKT/e NOS/NO mediated by Adipo R.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R587.1

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相關(guān)期刊論文 前1條

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