Puerarin partly counteracts the inflammatory response after cerebral ischemia/reperfusion via activating the cholinergic anti-inflammatory pathway

Puerarin, a major isoflavonoid derived from the Chinese medical herb radix puerariae （Gegen）, has been reported to inhibit neuronal apoptosis and play an anti-inflammatory role in focal cerebral ischemia model rats. Recent findings regarding stroke pathophysiology have recognized that anti-inflammation is an important target for the treatment of ischemic stroke. The cholinergic anti-inflammatory pathway is a highly robust neural-immune mechanism for inflammation control. This study was to investigate whether activating the cholinergic anti-inflammatory pathway can be involved in the mechanism of inhibiting the inflammatory response during puerarin-induced cerebral ischemia/reperfusion in rats. Results showed that puerarin pretreatment （intravenous injection） re- duced the ischemic infarct volume, improved neurological deficit after cerebral ischemia/reperfusion and decreased the levels of interleukin-1β, interleukin-6 and tumor necrosis factor-a in brain tissue. Pretreatment with puerarin （intravenous injection） attenuated the inflammatory response in rats, which was accompanied by janus-activated kinase 2 （JAK2） and signal transducers and activators of transcription 3 （STAT3） activation and nuclear factor kappa B （NF-KB） inhibition. These observa- tions were inhibited by the alpha7 nicotinic acetylcholine receptor （a7nAchR） antagonist a-bungarotoxin （a-BGT）. In addition, puerarin pretreatment increased the expression of a7nAchR mRNA in ischemic cerebral tissue. These data demonstrate that puerarin pretreatment strongly protects the brain against cerebral ischemia/reperfusion injury and inhibits the inflammatory re- sponse. Our results also indicated that the anti-inflammatory effect of puerarin may partly be medi- ated through the activation of the cholinergic anti-inflammatory pathway.

白藜芦醇对糖尿病白内障大鼠正沉默信息调节因子2相关酶1基因表达的影响

目的探讨白藜芦醇对糖尿病白内障大鼠晶状体正沉默信息调节因子2相关酶1(SIRT1)基因表达的影响。方法 80只Wistar大鼠随机分为4组:正常对照组、糖尿病模型组及白藜芦醇低剂量组及白藜芦醇高剂量组。糖尿病模型组、白藜芦醇低剂量组及白藜芦醇高剂量组一次性腹腔注射链脲菌素(STZ)(60 mg/kg)制备糖尿病大鼠模型。成模后低剂量组每日20 mg/kg,高剂量组每日100 mg/kg予白藜芦醇灌胃。裂隙灯显微镜照相机记录各组晶状体变化。12周实验结束时测定各组大鼠晶状体丙二醛(MDA)、超氧化物岐化酶(SOD)、谷胱甘肽过氧化酶含量。逆转录-聚合酶链反应(RT-PCR)检测各组大鼠晶状体SIRT1、肿瘤抑制因子P53、叉头转录因子1、核转录因子κB(NF-κB)表达情况。结果糖尿病模型组大鼠晶状体均发生不同程度混浊,甚至完全混浊,形成白内障。白内障糖尿病大鼠晶状体MDA(7.96±0.51)nmol/mg·prot较正常对照组(4.21±0.27)nmol/mg·prot升高(P<0.01),糖尿病白内障大鼠晶状体SOD、谷胱甘肽过氧化酶[(31.92±5.03)和(7.43±1.53)u/mg·prot]较正常对照组[(61.86±6.17)和(13.61±2.27)u/mg·prot]降低(P<0.01)。高剂量白藜芦醇组大鼠晶状体MDA(4.64±0.42)nmol/mg·prot较白内障糖尿病大鼠晶状体(7.96±0.51)nmol/mg·prot降低(P<0.01)。高剂量白藜芦醇组大鼠晶状体SOD、谷胱甘肽过氧化酶[(52.41±6.54)和(12.76±1.72)u/mg·prot]较白内障糖尿病大鼠晶状体SOD、谷胱甘肽过氧化酶[(31.92±5.03)和(7.43±1.53)u/mg·prot]增高(P<0.01)。白内障糖尿病大鼠晶状体SIRT1 mRNA表达(0.187±0.034)较正常对照组大鼠(0.523±0.089)降低(P<0.01)。高剂量白藜芦醇组大鼠晶状体SIRT1 mRNA表达(0.497±0.072)较白内障糖尿病大鼠晶状体(0.187±0.034)增强(P<0.01)。白内障糖尿病大鼠晶状体P53、叉头转录因子1、NF-κB mRNA表达(0.816±0.153)、(1.269±0.231)、(0.896±0.029)较正常对照组(0.592±0.104)、(0.674±0.112)、(0.495±0.008)增强(P<0.01)。高剂量白藜芦醇组糖尿病大鼠晶状体P53、叉头转录因子1、NF-κB mRNA表达(0.609±0.107)、(0.713±0.121)、(0.397±0.018)较白内障糖尿病大鼠(0.816±0.153)、(1.269±0.231)、(0.896±0.029)降低(P<0.01)。结论白藜芦醇可能通过调节SIRT1表达,进一步调节下游基因P53、叉头转录因子1、NF-κB的表达,抑制和延缓晶状体上皮细胞凋亡,延缓糖尿病大鼠白内障的发生发展。

盘龙七片调控SIRT1/NF-κB通路对骨关节炎软骨细胞凋亡的影响

目的观察盘龙七片对骨关节炎(osteoarthritis,OA)小鼠沉默信息转录调控因子(silent information regulator type 1,SIRT1)/核因子-κB(nuclear factor-kappa B,NF-κB)通路及软骨细胞凋亡的影响。方法手术切除右膝关节内侧半月板和前交叉韧带复制小鼠OA模型。将小鼠分为正常对照组,模型组,阳性对照组,盘龙七片低、中、高剂量组。通过番红O-快速绿染色观察膝关节结构变化,进行Mankin评分评估关节炎严重程度,流式细胞术检测软骨细胞凋亡指数,qRT-PCR检测Bax、Bcl-2、Caspase-3 mRNA表达水平,Western blot法检测软骨细胞SIRT1、NF-κB蛋白表达水平。结果与正常对照组比较,模型组小鼠膝关节软骨细胞凋亡率增加,Bax、Caspase-3 mRNA和NF-κB蛋白表达水平显著增加,Bcl-2 mRNA和SIRT1蛋白表达水平显著降低,差异均有统计学意义(P<0.05);与模型组比较,阳性对照组与盘龙七片低、中、高剂量组小鼠膝关节软骨细胞凋亡率显著降低,Bax、Caspase-3 mRNA和NF-κB蛋白表达水平显著降低,Bcl-2 mRNA和SIRT1蛋白表达水平显著增加,差异均有统计学意义(P<0.05),盘龙七片的作用呈明显的剂量依赖性。结论盘龙七片可抑制OA小鼠膝关节软骨细胞凋亡,其作用机制可能与调节SIRT1/NF-κB信号通路相关。

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.

化瘀消肿汤对深静脉血栓形成大鼠炎症反应的影响

目的探讨化瘀消肿汤(HXD)对深静脉血栓形成(DVT)大鼠炎症反应的影响。方法将雄性SD大鼠分为对照组(CK组)、模型组(Model组)、HXD低剂量组(HXD-L组,HXD 10.86 mg/kg)、HXD中剂量组(HXD-M组,HXD 21.71 mg/kg)、HXD高剂量组(HXD-H组,HXD 32.57 mg/kg)、阳性对照组(LMWHS组,低分子量肝素钠600 IU/kg)、沉默信息调节因子2(SIRT2)抑制剂组(AK-7组,AK-720 mg/kg)、HXD-M联合AK-7组(HXD-M+AK-7组,HXD 21.71 mg/kg+AK-720 mg/kg),每组12只。除CK组外,其余组大鼠均采用Reyers法构建DVT大鼠模型;建模后,各药物组分别灌胃/腹腔注射相应药液,每天1次,持续2周。末次给药24 h后,检测大鼠凝血功能指标[活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、凝血酶原时间(PT)、纤维蛋白原(FIB)]和血清、下腔静脉组织中炎症指标[白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)],观察其血栓形成情况并称定血栓湿重、干重,检测下腔静脉组织中组织因子(TF)、SIRT2蛋白的表达情况和核因子κB(NF-κB)p65的磷酸化、乙酰化水平。结果与CK组比较,Model组大鼠的APTT、TT、PT均显著缩短(P<0.05);FIB含量,IL-1β、IL-6、TNF-α水平,静脉血栓湿重、干重,TF蛋白染色评分,NF-κB p65蛋白的磷酸化、乙酰化水平均显著升高(P<0.05);下腔静脉血管内充满血栓,SIRT2蛋白的表达水平显著降低(P<0.05)。与Model组比较,HXD-L组、HXD-M组、HXD-H组和LMWHS组大鼠上述指标均显著改善,且HXD-M组、HXD-H和LMWHS组的效果显著优于HXD-L组(P<0.05);AK-7组大鼠上述指标的变化趋势与之相反,且AK-7可减弱中剂量HXD对模型大鼠炎症反应的抑制作用(P<0.05)。结论HXD可能通过激活SIRT2/NF-κB信号通路来抑制DVT大鼠的炎症反应。