替米沙坦对1型糖尿病大鼠睾丸脂联素及其受体表达的影响
发布时间：2018-09-10 13:43
【摘要】：目的:通过观察替米沙坦(telmisartan)对1型糖尿病(T1DM)大鼠睾丸脂联素(Adiponectin,APN)、脂联素受体(Adipo R)和腺苷酸活化蛋白激酶(AMPK)和蛋白激酶B/内皮性一氧化氮合酶/一氧化氮(Akt/e NOS/NO)表达的影响,探讨替米沙坦如何保护T1DM大鼠睾丸。方法:雄性SD大鼠27只随机分为正常对照组NC组8只和糖尿病造模组19只。用链脲佐菌素(STZ)诱导T1DM模型,16只成功。造模成功的大鼠被随机分为糖尿病组DM组8只和替米沙坦治疗组DT组8只。给DT组替米沙坦灌胃,NC组和DM组等量生理盐水灌胃。替米沙坦治疗8周后留取血标本,用于检测相关指标,然后处死动物并取出双侧睾丸,同时剪下附睾制备精子悬液。用40%甲醛溶液固定部分睾丸,HE染色;其余睾丸经液氮冷冻透后于-70℃冰箱待用。放免法测胰岛素(Insulin)及性激素。酶联免疫吸附(ELISA)法测脂联素(APN)、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)。用实时荧光定量PCR测Adipo R m RNA的表达。脂联素及其受体、AMPK、P-AMPK、AKT、P-AKT和e-NOS蛋白表达用免疫印迹法(Western Blot)测。结果:1.各组大鼠的体重、睾丸指数、精子数目、精子活动率的比较DM组较NC组大鼠睾丸重量明显减轻(P0.05),睾丸指数无明显变化(P0.05),精子数目、精子活动率明显降低(P0.05)。8周telmisartan治疗后,DT组大鼠睾丸重量、睾丸指数无明显变化(P0.05),精子数目、精子活动率明显升高(P0.05)。2.各组大鼠血糖、TCH、TG、FFA、性激素、Insulin、APN、睾丸NO、IL-6、TNF-α表达的比较与NC组比较,DM组大鼠血糖、TCH、TG、FFA、睾丸组织NO、IL-6、TNF-α均明显升高(P0.05),血清及睾丸组织睾酮(T)、血清及睾丸组织脂联素(APN)、胰岛素(Insulin)显著降低(P0.05),LH和FSH差异无统计学意义(P0.05)。8周替米沙坦治疗后,DT组大鼠血糖、TCH、TG、FFA、NO、IL-6、TNF-α均明显降低(P0.05),血清及睾丸组织APN明显上升(P0.05),血清及睾丸组织T、Insulin、LH、FSH无明显变化(P0.05)。3.各组大鼠睾丸组织形态学观察比较NC组睾丸组织生精小管及细胞排列正常,管腔内精子较多。DM组睾丸组织曲细精管萎缩变形,管腔内空泡状变性明显,生精细胞数量明显减少,生精小管之间的间质组织比例增加,但间质细胞体积缩小,胞浆减少,核固缩,间质组织中微血管腔闭塞、淤血,炎性细胞浸润于睾丸间质。DT组睾丸组织变性较DM组明显好转。4.各组大鼠睾丸脂联素受体1、2 m RNA的表达水平比较与NC组比较,DM组大鼠睾脂联素受体1 m RNA表达降低[(0.54±0.14)对(0.71±0.15),P0.05],脂联素受体2 m RNA表达无明显变化[(0.67±0.11)对(0.77±0.16),P0.05]。与DM组比较,DT组大鼠脂联素受体1和2 m RNA表达均无明显变化[(0.61±0.07)对(0.54±0.14)、(0.55±0.09)对(0.67±0.11),P0.05]。5.睾丸脂联素及其受体、P-AMPK/AMPK、P-AKT/Akt、e NOS蛋白表达水平的比较DM组较NC组大鼠睾丸脂联素、脂联素受体1、P-AMPK/AMPK蛋白表达均明显降低[(0.66±0.09)对(1.00±0.00),(0.68±0.05)对(1.00±0.00),(0.34±0.11)对(1.00±0.00),均P0.05],P-AKT/AKT和e NOS蛋白表达显著增高[(1.54±0.27)对(1.00±0.00),(1.56±0.26)对(1.00±0.00),P0.05],Adipo R2蛋白表达无明显变化[(1.02±0.13)对(1.00±0.00),P0.05]。替米沙坦治疗后,DT组大鼠睾丸脂联素和脂联素受体1蛋白表达显著增高[(0.84±0.09)对(0.66±0.09),(0.80±0.07)对(0.68±0.05),P0.05],P-AKT/AKT和e NOS表达蛋白显著降低[(1.24±0.39)对(1.54±0.27),(1.16±0.47)对(1.56±0.26),P0.05],而P-AMPK/AMPK和脂联素受体2蛋白表达无明显变化[(0.65±0.52)对(0.34±0.11),(1.02±0.15)对(1.02±0.13),P0.05]。结论:替米沙坦通过对APN、Adipo R的调节以及Adipo R介导的信号通路AKT/e NOS/NO的影响减轻糖尿病大鼠睾丸组织损伤程度,对T1DM大鼠睾丸组织产生保护的作用。
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1
本文编号：2234612
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1
【参考文献】
相关期刊论文 前1条
1 赵红光,朴春南,龚守良;糖尿病雄性生殖损伤及其发生机制[J];中华男科学杂志;2004年10期
，本文编号：2234612
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