白藜芦醇调控SIRT1/STAT1通路影响肝细胞癌恶性生物学过程

目的探讨白藜芦醇对肝细胞癌的作用及其相关机制,以期为肝细胞癌的治疗提供新思路。方法将HepG2细胞分为对照组、低剂量白藜芦醇组(RES‐L,10μmol/L)、高剂量组白藜芦醇(RES‐H,30μmol/L)和紫杉醇组(20μmol/L),通过MTT实验、集落形成实验、Transwell实验和细胞划痕实验分析白藜芦醇对HepG2细胞的活性、侵袭和迁移能力以及凋亡率的影响;采用实时荧光定量PCR法和蛋白免疫印迹法分析白藜芦醇对SIRT1/STAT1信号通路的影响。结果与对照组比较,白藜芦醇能够抑制HepG2细胞的活性[对照组、RES‐L组、RES‐H组、紫杉醇组分别为(100.875.78)%、(81.283.25)%、(46.368.05)%、(44.687.11)%],减少HepG2细胞集落形成数量[对照组、RES‐L组、RES‐H组、紫杉醇组分别为(102.526.82)%、(88.365.15)%、(30.266.05)%、(26.385.31)%],还能够抑制HepG2细胞的侵袭和迁移能力,促进HepG2细胞凋亡[对照组、RES‐L组、RES‐H组、紫杉醇组分别为(8.32±0.72)%、(12.16±3.05)%、(26.13±1.25)%、(41.88±6.81%)%]。另外,与对照组比较,白藜芦醇处理后,SIRT1和STAT1蛋白表达水平增加。结论白藜芦醇能够抑制HepG2细胞的恶性生物学过程,并且白藜芦醇的这一作用可能与激活SIRT1/STAT1信号通路有关。

白藜芦醇对KKAy小鼠体质量和脂肪分布的影响及机制

目的:观察白藜芦醇(resveratrol,res)对KKAy小鼠体质量和脂肪分布的影响,并探讨其中机制。方法:将36只8周龄KKAy小鼠分成3组,其中两组KKAy小鼠分别饲喂添加白藜芦醇(以饲料质量2、4g/kg添加)的AIN93G饲料,另外以12只同周龄C57BL/6J小鼠为对照。观察干预12周后白藜芦醇对小鼠体质量及皮下脂肪、附睾脂肪分布的影响;Western blot实验检测不同组小鼠附睾及皮下脂肪组织中沉默信息调节因子1(silent information regulator 1,sirt-1)、过氧化物增殖物受体γ(peroxisome proliferator-activated receptorγ,PPARγ)和CCAAT增强子结合蛋白α(CCAAT/enhancer-binding proteinα,C/EBPα)蛋白表达水平。结果:白藜芦醇干预可以在不影响摄食量的情况下减轻KKAy小鼠体质量,与模型组相比,高剂量res组小鼠体质量降低,且脂肪组织的质量及与脂肪组织系数降低;附睾及皮下脂肪组织中sirt-1的表达升高,PPARγ及C/EBPα蛋白表达水平降低。结论:白藜芦醇可以减轻肥胖小鼠体质量,降低体内脂肪含量,这可能与白藜芦醇提高脂肪组织中sirt-1的表达,从而抑制与脂肪合成及脂肪细胞分化相关蛋白有关。

Mir-217调节Sirt1/HIF-1α通路来调控肾小球系膜细胞炎性反应及纤维化过程

探讨Micro RNA-217(Mir-217)、低氧诱导因子1α(HIF-1α)及沉默信息调节因子1(Sirt1)在大鼠肾小球系膜细胞炎性反应及纤维化发生过程中的作用。用高糖培养的大鼠肾小球系膜细胞,采用RT-PCR技术检测Mir-217、HIF-1αm RNA及Sirt1的m RNA表达水平,采用Western-blotting技术检测HIF-1α、Sirt1、纤连蛋白(FN)、内皮素1(ET-1)、结缔组织生长因子(CTGF)表达水平,采用ELISA法检测血管内皮生长因子(VEGF)及转化生长因子-β1(TGF-β1)表达水平。结果显示,高糖刺激可下调大鼠肾小球系膜细胞中的Sirt1的表达水平,上调Mir-217、HIF-1α、FN、ET-1、CTGF、VEGF及TGF-β1的表达水平,差异具有统计学意义(p<0.05);而采用Sirt1特异性激活剂白藜芦醇预处理或Mir-217基因沉默可上调Sirt1表达水平,下调Mir-217、HIF-1α、FN、ET-1、CTGF、VEGF及TGF-β1的表达水平,差异具有统计学意义(p<0.05);因此,本研究认为Mir-217可以通过调节Sirt1/HIF-1α通路来调控肾小球系膜细胞炎性反应及纤维化过程。

Novel nervous and multi-system regenerative therapeutic strategies for diabetes mellitus with mTOR

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.

Effects of Manshenkangning Prescription on Adenine-induced Renal Interstitial Fibrosis in Rats

[Objectives]To study the protective effects of Manshenkangning Prescription on adenine-induced renal interstitial fibrosis in rats,and explore the possible mechanism.[Methods]Sixty Wistar male rats were divided into normal group,model group,control group(administered with 10 mg/(kg·d)losartan)and high,medium and low dose experimental groups(30,15,7.5 mg/(kg·d)Manshenkangning).The rat models of renal interstitial fibrosis were induced by intragastric administration of adenine(250 mg/(kg·d)).After 2 h,the above drugs were administered intragastrically for 21 consecutive days and the administration time was 30 consecutive days.Serum creatinine(SCr),blood urea nitrogen(BUN),24 h urinary protein(24 h MTP)and glomerular filtration rate(eGFR)were measured by biochemical method;renal histopathological changes were observed by hematoxylin-eosin(HE)staining.Renal collagen deposition in rats was observed by Masson staining.[Results]The SCr in model group and the high,medium and low dose experimental groups were(340.00±22.99),(176.80±18.60),(234.75±13.59),(266.11±14.78)μmol/L,and BUN were(23.74±2.51),(14.53±2.25),(18.78±0.88),(18.90±2.14)mmol/L;24 h MTP were(675.86±74.58),(323.81±41.83),(438.84±34.69),(493.76±37.04)mg/d;eGFR were(19.30±2.48),(49.96±10.95),(32.61±10.75),(27.18±5.98)mL/min,and the difference was statistically significant compared with the normal group(all P<0.05).HE staining and Masson staining showed that compared with normal group,the renal interstitial lesions in model group were severe and the renal interstitial collagen material was deposited in a large amount.The renal interstitial tubule injury was relieved and the renal interstitial collagen deposition was reduced in experimental groups.And the difference was statistically significant(all P<0.01).[Conclusions]Manshenkangning can significantly protect the kidney against the progress of interstitial fibrosis in rats.Its possible mechanism is to regulate the activity of SIRT1 and inhibit the expression of COX-2 in order to resist the inflammatory reaction of kidney and improve the ability of anti-oxidative stress of kidney,thus delaying the occurrence and development of chronic renal failure.

针刺对急性脑缺血大鼠沉默信息调节因子2相关酶1和核转录因子-κB蛋白的影响

目的:观察针刺对急性脑缺血大鼠缺血脑组织沉默信息调节因子2相关酶1(SIRT 1)和核转录因子-κB(NF-κB)的影响,探讨针刺治疗脑缺血的可能机制。方法:SD大鼠100只,随机均分为正常组、假手术组、模型组、非穴位组、穴位组,每组20只。采用开颅电凝大脑中动脉法复制急性脑缺血模型,穴位组针刺"百会"和"水沟"穴,非穴位组针刺"百会"和"水沟"穴旁开5mm处,每次30min,共治疗2次。TTC染色法测定大脑梗死面积,放射免疫法检测血清及缺血脑组织中白介素(IL)-1β、IL-6和IL-8含量,免疫印迹法检测缺血脑组织SIRT 1和NF-κB p 65蛋白的表达。结果:与正常组比较,模型组大鼠脑梗死范围明显,血清及缺血脑组织IL-1β、IL-6和IL-8含量显著升高,缺血脑组织SIRT 1蛋白表达减少,NF-κB p 65蛋白表达增加,差异具有统计学意义(均P<0.01)。与模型组相比,穴位组大鼠梗死区域变小,血清及缺血脑组织IL-1β、IL-6和IL-8含量降低,缺血脑组织SIRT 1蛋白表达升高,NF-κB p 65蛋白表达降低,差异具有统计学意义(均P<0.05)。结论:针刺能有效调节急性脑缺血大鼠炎性损伤,显著降低血清及缺血脑组织IL-1β、IL-6和IL-8含量,可能与调节SIRT 1/NF-κB通路有关。