中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2015年
8期
507-512
,共6页
闫旭红%郭志新%吴杰萍%王蕾%景晓锐%刘佳
閆旭紅%郭誌新%吳傑萍%王蕾%景曉銳%劉佳
염욱홍%곽지신%오걸평%왕뢰%경효예%류가
糖尿病,1型%脂联素%睾丸%大鼠%胰岛素
糖尿病,1型%脂聯素%睪汍%大鼠%胰島素
당뇨병,1형%지련소%고환%대서%이도소
Diabetes mellitus,type 1%Adiponectin%Testes%Rats%Insulin
目的：探讨胰岛素对1型糖尿病大鼠睾丸脂联素、脂联素受体及其介导的信号转导通路表达的影响。方法30只6周龄雄性SD大鼠（体重180～200 g）以随机数字表法分为正常对照组（A组，n=8)和链脲佐菌素注射组（n=22)。链脲佐菌素用于制备1型糖尿病大鼠模型。造模成功的16只雄性SD大鼠随机分为糖尿病组（B组，n=8)和糖尿病胰岛素治疗组(C组，n=8)。C组大鼠每日皮下注射精蛋白锌胰岛素3～5 U，使血糖控制于10 mmol/L左右，A、B组大鼠每日皮下注射等量的生理盐水。于实验8周末留取血标本，用于血生化指标、胰岛素、脂联素及性激素水平检测，然后处死动物，迅速取出双侧睾丸及附睾，计数精子数量及活动率，测定睾丸脂联素及其受体、单磷酸腺苷激活蛋白激酶(AMPK)、蛋白激酶B(AKT)、内皮型一氧化氮合酶(e?NOS)、一氧化氮(NO)、白细胞介素6（IL?6）和肿瘤坏死因子α(TNF?α)的表达水平。统计采用LSD?t检验和单因素方差分析。结果与A组比较，B组糖尿病大鼠睾丸组织出现显著病理改变，睾丸重量、精子数量及活动率均显著降低（t=3.899、21.139、26.770，均P<0.05）；睾丸组织表达脂联素及其受体1、p?AMPK/AMPK水平显著降低（A组均为1.00?0.00，B组分别为0.66?0.09、0.68?0.05、0.34?0.11，t=8.658、14.297、5.551，均P<0.05）；p?AKT/AKT、e?NOS蛋白表达水平显著升高(1.54?0.27比1.00?0.00、1.56?0.26比1.00?0.00，t=5.083、4.997，均P<0.05)。胰岛素治疗能显著逆转上述各项指标的变化。与B组相比，C组大鼠睾丸组织病变减轻，睾丸重量、精子数量及活动率降低升高（t=3.444、18.453、23.103,均P<0.05）；脂联素及其受体1、p?AMPK/AMPK表达水平显著升高(分别为0.85?0.11比0.66?0.09、0.82?0.06比0.68?0.05、0.87?0.40比0.34?0.11，t=4.969、6.151、4.454，均P<0.05)；p?AKT/AKT、e?NOS蛋白表达水平显著下降(1.31?0.25比1.54?0.27、1.24?0.29比1.56?0.26，t=2.169、2.830，均P<0.05)。结论脂联素及其受体介导的信号通路可能在1型糖尿病大鼠睾丸组织损伤中起重要作用；胰岛素可能通过调节该通路减轻组织损伤程度,对糖尿病大鼠睾丸组织产生保护作用。
目的：探討胰島素對1型糖尿病大鼠睪汍脂聯素、脂聯素受體及其介導的信號轉導通路錶達的影響。方法30隻6週齡雄性SD大鼠（體重180～200 g）以隨機數字錶法分為正常對照組（A組，n=8)和鏈脲佐菌素註射組（n=22)。鏈脲佐菌素用于製備1型糖尿病大鼠模型。造模成功的16隻雄性SD大鼠隨機分為糖尿病組（B組，n=8)和糖尿病胰島素治療組(C組，n=8)。C組大鼠每日皮下註射精蛋白鋅胰島素3～5 U，使血糖控製于10 mmol/L左右，A、B組大鼠每日皮下註射等量的生理鹽水。于實驗8週末留取血標本，用于血生化指標、胰島素、脂聯素及性激素水平檢測，然後處死動物，迅速取齣雙側睪汍及附睪，計數精子數量及活動率，測定睪汍脂聯素及其受體、單燐痠腺苷激活蛋白激酶(AMPK)、蛋白激酶B(AKT)、內皮型一氧化氮閤酶(e?NOS)、一氧化氮(NO)、白細胞介素6（IL?6）和腫瘤壞死因子α(TNF?α)的錶達水平。統計採用LSD?t檢驗和單因素方差分析。結果與A組比較，B組糖尿病大鼠睪汍組織齣現顯著病理改變，睪汍重量、精子數量及活動率均顯著降低（t=3.899、21.139、26.770，均P<0.05）；睪汍組織錶達脂聯素及其受體1、p?AMPK/AMPK水平顯著降低（A組均為1.00?0.00，B組分彆為0.66?0.09、0.68?0.05、0.34?0.11，t=8.658、14.297、5.551，均P<0.05）；p?AKT/AKT、e?NOS蛋白錶達水平顯著升高(1.54?0.27比1.00?0.00、1.56?0.26比1.00?0.00，t=5.083、4.997，均P<0.05)。胰島素治療能顯著逆轉上述各項指標的變化。與B組相比，C組大鼠睪汍組織病變減輕，睪汍重量、精子數量及活動率降低升高（t=3.444、18.453、23.103,均P<0.05）；脂聯素及其受體1、p?AMPK/AMPK錶達水平顯著升高(分彆為0.85?0.11比0.66?0.09、0.82?0.06比0.68?0.05、0.87?0.40比0.34?0.11，t=4.969、6.151、4.454，均P<0.05)；p?AKT/AKT、e?NOS蛋白錶達水平顯著下降(1.31?0.25比1.54?0.27、1.24?0.29比1.56?0.26，t=2.169、2.830，均P<0.05)。結論脂聯素及其受體介導的信號通路可能在1型糖尿病大鼠睪汍組織損傷中起重要作用；胰島素可能通過調節該通路減輕組織損傷程度,對糖尿病大鼠睪汍組織產生保護作用。
목적：탐토이도소대1형당뇨병대서고환지련소、지련소수체급기개도적신호전도통로표체적영향。방법30지6주령웅성SD대서（체중180～200 g）이수궤수자표법분위정상대조조（A조，n=8)화련뇨좌균소주사조（n=22)。련뇨좌균소용우제비1형당뇨병대서모형。조모성공적16지웅성SD대서수궤분위당뇨병조（B조，n=8)화당뇨병이도소치료조(C조，n=8)。C조대서매일피하주사정단백자이도소3～5 U，사혈당공제우10 mmol/L좌우，A、B조대서매일피하주사등량적생리염수。우실험8주말류취혈표본，용우혈생화지표、이도소、지련소급성격소수평검측，연후처사동물，신속취출쌍측고환급부고，계수정자수량급활동솔，측정고환지련소급기수체、단린산선감격활단백격매(AMPK)、단백격매B(AKT)、내피형일양화담합매(e?NOS)、일양화담(NO)、백세포개소6（IL?6）화종류배사인자α(TNF?α)적표체수평。통계채용LSD?t검험화단인소방차분석。결과여A조비교，B조당뇨병대서고환조직출현현저병리개변，고환중량、정자수량급활동솔균현저강저（t=3.899、21.139、26.770，균P<0.05）；고환조직표체지련소급기수체1、p?AMPK/AMPK수평현저강저（A조균위1.00?0.00，B조분별위0.66?0.09、0.68?0.05、0.34?0.11，t=8.658、14.297、5.551，균P<0.05）；p?AKT/AKT、e?NOS단백표체수평현저승고(1.54?0.27비1.00?0.00、1.56?0.26비1.00?0.00，t=5.083、4.997，균P<0.05)。이도소치료능현저역전상술각항지표적변화。여B조상비，C조대서고환조직병변감경，고환중량、정자수량급활동솔강저승고（t=3.444、18.453、23.103,균P<0.05）；지련소급기수체1、p?AMPK/AMPK표체수평현저승고(분별위0.85?0.11비0.66?0.09、0.82?0.06비0.68?0.05、0.87?0.40비0.34?0.11，t=4.969、6.151、4.454，균P<0.05)；p?AKT/AKT、e?NOS단백표체수평현저하강(1.31?0.25비1.54?0.27、1.24?0.29비1.56?0.26，t=2.169、2.830，균P<0.05)。결론지련소급기수체개도적신호통로가능재1형당뇨병대서고환조직손상중기중요작용；이도소가능통과조절해통로감경조직손상정도,대당뇨병대서고환조직산생보호작용。
Objective To explore the effect of insulin on the expression of adiponectin, adiponectin receptors, and adiponectin receptor?mediated signaling pathways in the testes of type 1 diabetic rats. Methods Thirty male 6?week Sprague?Dawley(SD) rats(body weight: 180?200 g) were randomly divided into normal control group (group A, n=8) and streptozotocin injection group(n=22) by using random number table. Type 1 diabetic model was established by intraperitoneal injection of streptozotocin. Sixteen successfully?induced diabetic rats were randomly divided into diabetic group(group B, n=8) and diabetic treated with insulin(group C, n=8). Rats in group B were given subcutaneous protamine?zinc insulin in does of 3?5 U daily to control blood glucose at 10 mmol/L. The other two groups were given equal volume of normal saline daily. At the end of the eight?week of experiment, rats were sacrificed after blood samples were <br> collected, and then the bilateral testes were collected quickly. Blood samples were used to detect the levels of biochemical index, insulin, adiponectin and sex hormone. Sperm number and motility were counted. The expression of testicular adiponectin, adiponectin receptors, adenosine 5′?monophosphate (AMP)?activated protein kinase(AMPK), protein kinase B(AKT), endothelial nitric oxide synthase(e?NOS), nitric oxide(NO), interleukin?6(IL?6), and tumor necrosis factor alpha(TNF?α) were assayed. Statistical analysis was performed by using LSD?t test and one way ANOVA. Results There were significant pathological changes in the testes of group B than those in group A. Compared with group A, the level of testes weight, sperm number and motility were decreased in group B(t=3.899, 21.139, 26.770, all P<0.05), the testicular adiponectin and its receptor 1, the ratio of p?AMPK to AMPK were significantly decreased in group B ((0.66?0.09) vs (1.00?0.00), (0.68?0.05) vs (1.00?0.00), (0.34?0.11) vs (1.00?0.00), t=8.658, 14.297, 5.551, respectively, all P<0.05);however, the level of the ratio of p?AKT to AKT, e?NOS in the testes were significantly increased in group B (1.54?0.27 vs 1.00?0.00, 1.56?0.26 vs 1.00?0.00, t=5.083, 4.997, respectively, both P<0.05). These changes could be significantly reversed by insulin treatment. Compared with group B, the level of testes weight, sperm number and motility were increased in group C (t=3.444, 18.453, 23.103, all P<0.05);the level of adiponectin and its receptor 1, the ratio of p?AMPK to AMPK were significantly increased in group C (0.85?0.11 vs 0.66?0.09, 0.82?0.06 vs 0.68?0.05, 0.87?0.40 vs 0.34?0.11, t=4.969, 6.151, 4.454, respectively, all P<0.05);surely, the level of the ratio of p?AKT to AKT, e?NOS were significantly decreased in group B than those in group A (1.31?0.25 vs 1.54?0.27, 1.24?0.29 vs 1.56?0.26, t=2.169, 2.830, respectively, both P<0.05). Conclusions Adiponectin and its receptors and adiponectin receptor?mediated signaling pathway may play an important role in testicular tissue injury in diabetic rats. Insulin may reduce the the degree of testicular tissue damage and produce the protective effect on testicular tissues in diabetic rats by regulating the expression of adiponectin, adiponectin receptors and adiponectin receptor?mediated signaling pathways.
