中药调节平滑肌细胞内质网应激防治动脉粥样硬化研究进展

血管平滑肌细胞(VSMC)在病理因素诱导下异常增殖、迁移、表型改变是动脉粥样硬化(AS)发病的关键环节。与内质网应激(ERS)相关的凋亡途径可能导致VSMC增殖、凋亡失衡,从而在AS发生和发展中起到重要作用。中药干预调节VSMC异常增殖和凋亡在AS的预防和治疗中发挥重要作用。综述近年来中药单体和复方调节VSMS ERS从而防治AS的部分研究进展,以期进一步阐释与总结中医中药防治AS的作用特点与可能机制。     

Ischemia-reperfusion injury up-regulates Pim-3 gene expression in myocardial tissue

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism.Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats.A total of 30 SD male adult rats were randomly divided into 5 groups:group A (sham operation,n=6);group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery,n=6);group C (in which the rats received 30 min of ischemia,n=6),group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min,n=6 in each).The left ventricular tissues were removed immediately after the ischemia-reperfusion injury.Neonatal cardiomyocytes were cultured and treated with different concentrations of H 2 O 2 (0,5,10,20 μmol/L) or tumor necrosis factor-α (TNF-α,0,1,5,10 ng/mL).The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR,western blotting and immunohistochemistry.Additionally,neonatal cardiomyocytes were transfected with Pim-3 siRNA,and induced to develop apoptosis by using H 2 O 2.The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein.Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues.Furthermore,H 2 O 2 but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes.And Pim-3 silencing failed to strengthen the H 2 O 2-inducing apoptosis in cardiomyocytes.It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.     

中药调控干细胞Notch信号通路的研究进展

Notch信号通路是一条传导途经高度保守的信号通路,是调控干细胞自我更新、分化、发育和凋亡的重要通路之一。中药调控Notch通路影响干细胞的增殖和分化已成为该领域研究热点。本综述阐述了中药通过调控Notch通路来影响骨髓间充质干细胞(BMSCs)、肿瘤干细胞(CSCs)、神经干细胞(NSCs)、胚胎干细胞等干细胞增殖、分化和凋亡,为中药的临床应用提供依据,也为研究中药调节干细胞的作用机制提供思路和方法。     

PI3K-like kinases restrain Pim gene expression in endothelial cells

Pim kinases contribute to tumor formation and development of lymphoma,which shows enhanced DNA replication,DNA recombination and repair.Endothelial cells (ECs) express all the three members of Pim kinase gene family.We hypothesized that DNA repair gene would regulate Pim ex-pression in ECs.Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium.The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining.The siRNA fragments were synthesized and transfected by using Lipofectamine LTX.The total cellular RNA was extracted from the cells by using Trizol reagent.cDNAs were quantified by semi-quantity PCR.The effects of LY294002 and wortmannin on RNA stability in ECs were also ex-amined.Our data showed that LY294002 and wortmannin,phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors,increased Pim mRNA expression in ECs without altering the mRNA stabil-ity.RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1,respectively.Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs.But etoposide,a nucleoside analogue,which could activate DNA-PKcs and ATM,increased Pim expression in ECs.Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.     

高糖对ECV304细胞中Cofilin-1表达及PKC活性的影响

目的:探讨高糖刺激对ECV304细胞中Cofilin-1的表达及蛋白激酶C(PKC)活性的影响。方法:常规培养ECV304细胞,分组给药后,非放射性酶法检测该细胞中PKC的活性,RT-PCR法检测细胞Cofilin-1总mRNA的表达。结果:经高糖作用后,ECV304细胞PKC活性较低糖的正常组明显增加(P<0.01),细胞内Cofilin-1总mRNA的表达显著增强(P<0.01);而预先给予PKC抑制剂GF109203x处理能显著地抑制高糖介导的ECV304细胞内PKC活性的升高(P<0.01)和胞内Cofilin-1总mRNA的表达(P<0.05),但对正常组PKC活性和Cofilin-1总mRNA的表达无显著性影响。结论:高糖刺激可能通过PKC信号传导通路使Cofilin-1表达增强,进一步导致糖尿病血管病变的发生。     

Pim-3在大鼠脂肪组织中的表达及其与脂肪胰岛素抵抗的关系

目的: 检测Pim-3在脂肪中的表达,并探讨其在脂肪的胰岛素抵抗发生过程中的作用。方法: RT-PCR法检测脂肪组织中Pim-3 mRNA的表达,免疫荧光化学法检测Pim-3蛋白表达及亚细胞定位;检测胰岛素抵抗大鼠的附睾及肾周脂肪组织中Pim-3 mRNA表达变化;体外诱导大鼠骨髓间充质干细胞分化为脂肪细胞,油红O染色判断分化程度,并观察分化前后Pim-3表达变化。RT-PCR法检测高胰岛素(100 nmol/L)对骨髓间充质干细胞来源的脂肪细胞中Pim-3表达变化的影响。结果: (1) 脂肪组织中具有较高水平的Pim-3 mRNA表达,免疫荧光结果显示其蛋白主要分布于胞浆;(2)胰岛素抵抗大鼠附睾及肾周脂肪组织中Pim-3 mRNA表达显著低于对照组 (P<0.05) ;(3)大鼠骨髓间充质干细胞成功分化为脂肪细胞,分化后Pim-3 mRNA表达水平明显增高;(4)高胰岛素处理后,脂肪细胞中Pim-3 mRNA表达显著降低。结论: Pim-3表达于脂肪组织中,并可能参与骨髓间充质干细胞来源的脂肪细胞分化过程及脂肪组织胰岛素抵抗的发病过程。