中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
22期
1773-1778
,共6页
王蕾%郭志新%吴杰萍%闫旭红%景晓锐%刘佳
王蕾%郭誌新%吳傑萍%閆旭紅%景曉銳%劉佳
왕뢰%곽지신%오걸평%염욱홍%경효예%류가
糖尿病,1型%睾丸%脂联素
糖尿病,1型%睪汍%脂聯素
당뇨병,1형%고환%지련소
Diabetes mellitus,type 1%Testis%Adiponectin
目的 观察替米沙坦对1型糖尿病大鼠睾丸脂联素、脂联素受体及其信号转导途径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,探讨替米沙坦对1型糖尿病大鼠睾丸保护作用及机制.方法 将27只雄性SD大鼠用随机数字表法分为正常对照(NC)组(8只)和糖尿病造模组(19只).采用链脲佐菌素(STZ)诱导1型糖尿病大鼠模型,共有16只大鼠造模成功.将造模成功的糖尿病大鼠随机分为糖尿病(DM)组(8只)和替米沙坦治疗(DT)组(8只).DT组予以替米沙坦灌胃,NC组和DM组给予等量生理盐水灌胃.替米沙坦组治疗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.部分睾丸组织置于中性40％甲醛溶液固定,用于HE染色,其余睾丸组织经液氮冷冻透后保存于-70℃冰箱备用.胰岛素及性激素用放射免疫法测定.脂联素、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)用酶联免疫吸附(ELISA)法测定,一氧化氮(NO)用硝酸还原酶法测定.脂联素及其受体mRNA表达用实时荧光定量PCR检测,脂联素及其受体、AMPK、磷酸化腺苷酸活化蛋白激酶(P-AMPK)、AKT、磷酸化蛋白激酶B(P-AKT)和e-NOS蛋白表达用免疫印迹法测定.结果 与NC组比较,DM组大鼠睾丸组织出现显著病理改变,P-AKT/AKT、e-NOS和NO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)、(1.75±0.28)比(1.08±0.02)μmol/g,均P＜0.05],血清脂联素水平[(622.46±95.86)比(2 022.07±51.13) ng/ml,P＜0.05],睾丸脂联素、脂联素受体1、P-AMPK/AMPK蛋白表达水平显著降低[(0.66±0.09)比(1.00±0.00)、(0.68±0.05)比(1.00±0.00)、(0.34±0.11)比(1.00 ±0.00),均P＜0.05].脂联素受体2蛋白表达水平无明显变化(1.02±0.13比1.00±0.00,P＞0.05).替米沙坦治疗8周后,DT组大鼠睾丸组织病理改变减轻,P-AKT/AKT、e-NOS和NO表达水平显著降低[(1.24±0.39)比(1.54±0.27)、(1.16±0.47)比(1.56±0.26)、(1.35±0.30)比(1.75±0.28) μmol/g,均P＜0.05],血清脂联素水平[(1 051.55±102.55)比(622.46±95.86) ng/ml,P＜0.001]、睾丸脂联素、睾丸脂联素受体1蛋白表达水平显著升高[(0.84±0.09)比(0.66±0.09)、(0.80±0.07)比(0.68±0.05),均P＜0.05],而睾丸P-AMPK/AMPK、脂联素受体2蛋白表达水平无明显变化[(0.65±0.52)比(0.34±0.11)、(1.02±0.15)比(1.02±0.13),均P＞0.05].结论 替米沙坦通过调节脂联素、脂联素受体以及脂联素受体介导的信号通路Akt/e-NOS/NO减轻糖尿病大鼠睾丸组织损伤程度,对糖尿病大鼠睾丸组织产生保护作用.
目的 觀察替米沙坦對1型糖尿病大鼠睪汍脂聯素、脂聯素受體及其信號轉導途徑腺苷痠活化蛋白激酶(AMPK)和蛋白激酶B/內皮性一氧化氮閤酶/一氧化氮(Akt/e-NOS/NO)錶達的影響,探討替米沙坦對1型糖尿病大鼠睪汍保護作用及機製.方法 將27隻雄性SD大鼠用隨機數字錶法分為正常對照(NC)組(8隻)和糖尿病造模組(19隻).採用鏈脲佐菌素(STZ)誘導1型糖尿病大鼠模型,共有16隻大鼠造模成功.將造模成功的糖尿病大鼠隨機分為糖尿病(DM)組(8隻)和替米沙坦治療(DT)組(8隻).DT組予以替米沙坦灌胃,NC組和DM組給予等量生理鹽水灌胃.替米沙坦組治療8週後留取血標本,用于檢測相關指標,然後處死動物併取齣雙側睪汍,同時剪下附睪製備精子懸液.部分睪汍組織置于中性40％甲醛溶液固定,用于HE染色,其餘睪汍組織經液氮冷凍透後保存于-70℃冰箱備用.胰島素及性激素用放射免疫法測定.脂聯素、白細胞介素6(IL-6)和腫瘤壞死因子α(TNF-α)用酶聯免疫吸附(ELISA)法測定,一氧化氮(NO)用硝痠還原酶法測定.脂聯素及其受體mRNA錶達用實時熒光定量PCR檢測,脂聯素及其受體、AMPK、燐痠化腺苷痠活化蛋白激酶(P-AMPK)、AKT、燐痠化蛋白激酶B(P-AKT)和e-NOS蛋白錶達用免疫印跡法測定.結果 與NC組比較,DM組大鼠睪汍組織齣現顯著病理改變,P-AKT/AKT、e-NOS和NO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)、(1.75±0.28)比(1.08±0.02)μmol/g,均P＜0.05],血清脂聯素水平[(622.46±95.86)比(2 022.07±51.13) ng/ml,P＜0.05],睪汍脂聯素、脂聯素受體1、P-AMPK/AMPK蛋白錶達水平顯著降低[(0.66±0.09)比(1.00±0.00)、(0.68±0.05)比(1.00±0.00)、(0.34±0.11)比(1.00 ±0.00),均P＜0.05].脂聯素受體2蛋白錶達水平無明顯變化(1.02±0.13比1.00±0.00,P＞0.05).替米沙坦治療8週後,DT組大鼠睪汍組織病理改變減輕,P-AKT/AKT、e-NOS和NO錶達水平顯著降低[(1.24±0.39)比(1.54±0.27)、(1.16±0.47)比(1.56±0.26)、(1.35±0.30)比(1.75±0.28) μmol/g,均P＜0.05],血清脂聯素水平[(1 051.55±102.55)比(622.46±95.86) ng/ml,P＜0.001]、睪汍脂聯素、睪汍脂聯素受體1蛋白錶達水平顯著升高[(0.84±0.09)比(0.66±0.09)、(0.80±0.07)比(0.68±0.05),均P＜0.05],而睪汍P-AMPK/AMPK、脂聯素受體2蛋白錶達水平無明顯變化[(0.65±0.52)比(0.34±0.11)、(1.02±0.15)比(1.02±0.13),均P＞0.05].結論 替米沙坦通過調節脂聯素、脂聯素受體以及脂聯素受體介導的信號通路Akt/e-NOS/NO減輕糖尿病大鼠睪汍組織損傷程度,對糖尿病大鼠睪汍組織產生保護作用.
목적 관찰체미사탄대1형당뇨병대서고환지련소、지련소수체급기신호전도도경선감산활화단백격매(AMPK)화단백격매B/내피성일양화담합매/일양화담(Akt/e-NOS/NO)표체적영향,탐토체미사탄대1형당뇨병대서고환보호작용급궤제.방법 장27지웅성SD대서용수궤수자표법분위정상대조(NC)조(8지)화당뇨병조모조(19지).채용련뇨좌균소(STZ)유도1형당뇨병대서모형,공유16지대서조모성공.장조모성공적당뇨병대서수궤분위당뇨병(DM)조(8지)화체미사탄치료(DT)조(8지).DT조여이체미사탄관위,NC조화DM조급여등량생리염수관위.체미사탄조치료8주후류취혈표본,용우검측상관지표,연후처사동물병취출쌍측고환,동시전하부고제비정자현액.부분고환조직치우중성40％갑철용액고정,용우HE염색,기여고환조직경액담냉동투후보존우-70℃빙상비용.이도소급성격소용방사면역법측정.지련소、백세포개소6(IL-6)화종류배사인자α(TNF-α)용매련면역흡부(ELISA)법측정,일양화담(NO)용초산환원매법측정.지련소급기수체mRNA표체용실시형광정량PCR검측,지련소급기수체、AMPK、린산화선감산활화단백격매(P-AMPK)、AKT、린산화단백격매B(P-AKT)화e-NOS단백표체용면역인적법측정.결과 여NC조비교,DM조대서고환조직출현현저병리개변,P-AKT/AKT、e-NOS화NO표체수평현저승고[(1.54±0.27)비(1.00 ±0.00)、(1.56 ±0.26)비(1.00 ±0.00)、(1.75±0.28)비(1.08±0.02)μmol/g,균P＜0.05],혈청지련소수평[(622.46±95.86)비(2 022.07±51.13) ng/ml,P＜0.05],고환지련소、지련소수체1、P-AMPK/AMPK단백표체수평현저강저[(0.66±0.09)비(1.00±0.00)、(0.68±0.05)비(1.00±0.00)、(0.34±0.11)비(1.00 ±0.00),균P＜0.05].지련소수체2단백표체수평무명현변화(1.02±0.13비1.00±0.00,P＞0.05).체미사탄치료8주후,DT조대서고환조직병리개변감경,P-AKT/AKT、e-NOS화NO표체수평현저강저[(1.24±0.39)비(1.54±0.27)、(1.16±0.47)비(1.56±0.26)、(1.35±0.30)비(1.75±0.28) μmol/g,균P＜0.05],혈청지련소수평[(1 051.55±102.55)비(622.46±95.86) ng/ml,P＜0.001]、고환지련소、고환지련소수체1단백표체수평현저승고[(0.84±0.09)비(0.66±0.09)、(0.80±0.07)비(0.68±0.05),균P＜0.05],이고환P-AMPK/AMPK、지련소수체2단백표체수평무명현변화[(0.65±0.52)비(0.34±0.11)、(1.02±0.15)비(1.02±0.13),균P＞0.05].결론 체미사탄통과조절지련소、지련소수체이급지련소수체개도적신호통로Akt/e-NOS/NO감경당뇨병대서고환조직손상정도,대당뇨병대서고환조직산생보호작용.
Objective To explore the effects of telmisartan on the expressions of adiponectin and adiponectin receptor and its signal transduction pathway AMP-activated protein kinase (AMPK),Akt/ endothelial nitric oxide synthase (Akt/e-NOS/NO) and examine the possible protective mechanisms of telmisartan in testis of type 1 diabetic rats.Methods A total of 27 male Sprague-Dawley rats were randomly divided into normal control (NC,n =8) group and diabetic model (n =19) group.Diabetes was induced by an intraperitoneal injection of streptozotocin (STZ).And 16 established diabetic rats were randomly divided into diabetic control (DM,n =8) and diabetic treated with telmisartan (DT,n =8) groups.Group DT received a lavage of telmisartan while groups NC and DM had an equal volume of normal saline by gavage.At the end of 8-week of telmisartan treatment,the animals were sacrificed after harvesting of blood samples.Bilateral testes were extracted and epididymis was minced for preparing sperm suspension.Blood samples were used to detect the related parameters.Some tresticular tissues were fixed in neutral 40％ formaldehyde solution and processed for histological analysis.And the remaining testicular tissues were immediately placed into liquid nitrogen and then stored in a-70 ℃ refrigerator until analyses.The levels of insulin and sex hormone were detected by radioimmunoassay.The levels of adiponectin,interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were assayed by enzyme-linked immunosorbent assay (ELISA).The mRNA expression of testicular adiponectin and its receptor was assessed by real-time fluorescent quantitative polymerase chain reaction (PCR).The protein expressions of testicular adiponectin,adiponectin receptor,AMPK,P-AMPK,AKT,P-AKT and e-NOS were analyzed by Western blot.Results There was significant pathological changes in testes in group DM than that in group NC.The levels of P-AKT/AKT,e-NOS and NO were significantly increased in group DM than those in group NC [(1.54 ± 0.27) vs (1.00 ± 0.00),(1.56±0.26) vs(1.00±0.00),(1.75±0.28)vs(1.08±0.02)μmol/g,allP＜0.05].The levels of serum adiponectin,testicular adiponectin and its receptor 1,and the ratio of P-AMPK to AMPK significantly decreased in group DM than that in group NC [(622.46 ±95.86) vs (2 022.07 ±51.13)ng/ml,(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),all P＜ 0.05].There was no significant difference in the expression of adiponectin receptor 2 between group DM and NC[(1.02 ± 0.13) vs (1.00 ± 0.00),P ＞ 0.05].After 8-week telmisartan treatment,the pathological changes of testes became alleviated in diabetic rats.The levels of P-AKT/AKT,e-NOS and NO significantly decreased in group DT than those in group DM [(1.24 ±0.39) vs (1.54 ±0.27),(1.16 ±0.47) vs (1.56 ± 0.26),(1.35 ± 0.30) vs (1.75 ± 0.28) μmol/g,all P ＜ 0.05].The serum and testicular levels of adiponectin and testicular adiponectin receptor 1 significantly increased in group DT than those in group DM [(1 051.55 ± 102.55) vs (622.46 ±95.86),(0.84 ±0.09) vs (0.66 ±0.09),(0.80 ±0.07) vs (0.68 ± 0.05),all P ＜ 0.05].No significant difference existed in the ratio of P-AMPK to AMPK in testes or the expression of adiponectin receptor 2 between group DM and NC [(0.65 ± 0.52) vs (0.34 ± 0.11),(1.02 ± 0.15) vs (1.02 ± 0.13),all P ＞ 0.05].Conclusion Telmisartan may reduce the injury degree of testes and play protective roles in testicular tissues in diabetic rats by regulating the expressions of adiponectin,adiponectin receptor and the signal pathways mediated by adiponectin receptor.
