目的:通过观察丹参多酚酸盐对膜性肾病(MN)大鼠肾组织中腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子(Sirt1)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)蛋白的表达及细胞自噬和凋亡的情况,探讨其治疗MN的可能的分子机制。...目的:通过观察丹参多酚酸盐对膜性肾病(MN)大鼠肾组织中腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子(Sirt1)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)蛋白的表达及细胞自噬和凋亡的情况,探讨其治疗MN的可能的分子机制。方法:80只雄性SD大鼠随机分为正常组,模型组,盐酸贝那普利组(10 mg·kg^(-1)),丹参多酚酸盐低、中、高剂量组(16.7、33.3、66.7 mg·kg^(-1)),通过尾静脉注射阳离子化牛血清白蛋白(C-BSA)的方法造模。造模成功后,各组按照相应比例剂量连续给药4周后留取24 h尿、血清和肾组织,尿液用于检测24 h尿蛋白定量(24 h UTP)、血清用于检测血尿素氮(BUN)、血肌酐(SCr)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)的含量。采用光镜、电镜、免疫荧光法观察肾脏病理学变化,蛋白免疫印迹法(Western blot)检测大鼠肾组织磷酸化(p)-AMPK、AMPK、p-Sirt1、Sirt1、PGC-1α蛋白表达水平;免疫组化(IHC)检测大鼠肾组织自噬特异性基因-1(Beclin-1)、微管相关蛋白1轻链3(LC3)Ⅱ、泛素结合蛋白(p62)、B细胞淋巴瘤(Bcl)-2、Bcl-2相关X蛋白(Bax)、胱天蛋白酶(Caspase)-7蛋白表达水平。结果:与正常组比较,模型组大鼠24 h UTP、IL-6、TNF-α、CRP、MDA水平显著升高(P<0.01),SOD和GSH-Px水平显著降低(P<0.01),BUN、SCr变化差异无统计学意义;与模型组比较,丹参多酚酸盐低中高剂量组和贝那普利组大鼠24 h UTP、IL-6、TNF-α、CRP、MDA水平显著降低(P<0.01),SOD和GSH-Px水平显著升高(P<0.01)。在苏木素-伊红(HE)、马松(Masson)染色、免疫荧光及电镜下观察可见模型组大鼠肾组织病理损伤明显,贝那普利和丹参多酚酸盐治疗后,肾组织细胞的病理损伤逐渐改善。与正常组比较,模型组大鼠肾脏p-AMPK/AMPK、p-Sirt1/Sirt1、PGC-1α、Bcl-2、Beclin-1、LC3Ⅱ表达显著降低(P<0.01),Bax、Caspase-7、p62的表达显著增加(P<0.01);与模型组比较,贝那普利和丹参多酚酸盐治疗后大鼠肾脏p-AMPK/AMPK、p-Sirt1/Sirt1、PGC-1α、Bcl-2、Beclin-1、LC3Ⅱ表达显著升高(P<0.01),Bax、Caspase-7、p62的表达显著降低(P<0.01)。结论:丹参多酚酸盐对MN大鼠肾保护作用,这可能与激活AMPK/Sirt1/PGC-1α通路,上调自噬,减少凋亡有关。展开更多
Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and af...Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.展开更多
文摘目的:通过观察丹参多酚酸盐对膜性肾病(MN)大鼠肾组织中腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子(Sirt1)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)蛋白的表达及细胞自噬和凋亡的情况,探讨其治疗MN的可能的分子机制。方法:80只雄性SD大鼠随机分为正常组,模型组,盐酸贝那普利组(10 mg·kg^(-1)),丹参多酚酸盐低、中、高剂量组(16.7、33.3、66.7 mg·kg^(-1)),通过尾静脉注射阳离子化牛血清白蛋白(C-BSA)的方法造模。造模成功后,各组按照相应比例剂量连续给药4周后留取24 h尿、血清和肾组织,尿液用于检测24 h尿蛋白定量(24 h UTP)、血清用于检测血尿素氮(BUN)、血肌酐(SCr)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、丙二醛(MDA)的含量。采用光镜、电镜、免疫荧光法观察肾脏病理学变化,蛋白免疫印迹法(Western blot)检测大鼠肾组织磷酸化(p)-AMPK、AMPK、p-Sirt1、Sirt1、PGC-1α蛋白表达水平;免疫组化(IHC)检测大鼠肾组织自噬特异性基因-1(Beclin-1)、微管相关蛋白1轻链3(LC3)Ⅱ、泛素结合蛋白(p62)、B细胞淋巴瘤(Bcl)-2、Bcl-2相关X蛋白(Bax)、胱天蛋白酶(Caspase)-7蛋白表达水平。结果:与正常组比较,模型组大鼠24 h UTP、IL-6、TNF-α、CRP、MDA水平显著升高(P<0.01),SOD和GSH-Px水平显著降低(P<0.01),BUN、SCr变化差异无统计学意义;与模型组比较,丹参多酚酸盐低中高剂量组和贝那普利组大鼠24 h UTP、IL-6、TNF-α、CRP、MDA水平显著降低(P<0.01),SOD和GSH-Px水平显著升高(P<0.01)。在苏木素-伊红(HE)、马松(Masson)染色、免疫荧光及电镜下观察可见模型组大鼠肾组织病理损伤明显,贝那普利和丹参多酚酸盐治疗后,肾组织细胞的病理损伤逐渐改善。与正常组比较,模型组大鼠肾脏p-AMPK/AMPK、p-Sirt1/Sirt1、PGC-1α、Bcl-2、Beclin-1、LC3Ⅱ表达显著降低(P<0.01),Bax、Caspase-7、p62的表达显著增加(P<0.01);与模型组比较,贝那普利和丹参多酚酸盐治疗后大鼠肾脏p-AMPK/AMPK、p-Sirt1/Sirt1、PGC-1α、Bcl-2、Beclin-1、LC3Ⅱ表达显著升高(P<0.01),Bax、Caspase-7、p62的表达显著降低(P<0.01)。结论:丹参多酚酸盐对MN大鼠肾保护作用,这可能与激活AMPK/Sirt1/PGC-1α通路,上调自噬,减少凋亡有关。
基金supported by American Diabetes Association,American Heart Association,NIH NIEHS,NIH NIA,NIH NINDS,and NIH ARRA
文摘Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.
