Endoplasmic reticulum stress in insulin resistance and diabetes

The endoplasmic reticulum is the main intracellular Ca²⁺ store for Ca²⁺ release during cell signaling. There are different strategies to avoid ER Ca²⁺ depletion. Release channels utilize first Ca²⁺-bound to proteins and this minimizes the reduction of the free luminal [Ca²⁺]. However, if release channels stay open after exhaustion of Ca²⁺-bound to proteins, then the reduction of the free luminal ER [Ca²⁺] (via STIM proteins) activates Ca²⁺ entry at the plasma membrane to restore the ER Ca²⁺ load, which will work provided that SERCA pump is active. Nevertheless, there are several noxious conditions that result in decreased activity of the SERCA pump such as oxidative stress, inflammatory cytokines, and saturated fatty acids, among others. These conditions result in a deficient restoration of the ER [Ca²⁺] and lead to the ER stress response that should facilitate recovery of the ER. However, if the stressful condition persists then ER stress ends up triggering cell death and the ensuing degenerative process leads to diverse pathologies; particularly insulin resistance, diabetes and several of the complications associated with diabetes. This scenario suggests that limiting ER stress should decrease the incidence of diabetes and the mobility and mortality associated with this illness.     

Effect of Sodium Selenite and Vitamin E on the Renal Cortex in Rats: An Ultrastructure Study

This study examined the use of vitamin E to alleviate toxic effects of sodium selenite. Adult male albino rats (n = 50) was divided into five groups. Group 1 was control, Groups 2 and 4 were treated with sodium selenite (2 mg/kg) for 2 and 4 weeks, respectively, Groups 3 and 5 were treated with sodium selenite (2 mg/kg) and vitamin E (100 mg/kg) for 2 and 4 weeks, respectively. Renal tissues were studied using anti-BCL2 and examined ultrastructurally. Positive Bax immunoreactivity was detected after 2 and more positive after 4 weeks and nearly all groups improved with co-administration of vitamin E. Ultrastructural study revealed lesions in Bowman's capsule and proximal convoluted tubules. The submicroscopic study revealed damage and necrosis of cortical structures after 2 and 4 weeks, respectively. After 4 weeks, cellular changes were seen, such as vacuolation and moderate degeneration of cells, widening of the urinary space scattered through the cortex with loss of cellular details, formation of apical buds, degeneration, and cellular rupture. Present findings disclosed an ameliorative effect of adding vitamin E to sodium selenite-induced changes in cortical tissues. Clinically, it is advised to add vitamin E to avoid selenium overdose hazards.     

Pseudomonas aeruginosa ExoT Induces Mitochondrial Apoptosis in Target Host Cells in a Manner That Depends on Its GTPase-activating Protein (GAP) Domain Activity

Pseudomonas aeruginosa is the most common cause of hospital-acquired pneumonia and a killer of immunocompromised patients. We and others have demonstrated that the type III secretion system (T3SS) effector protein ExoT plays a pivotal role in facilitating P. aeruginosa pathogenesis. ExoT possesses an N-terminal GTPase-activating protein (GAP) domain and a C-terminal ADP-ribosyltransferase (ADPRT) domain. Because it targets multiple non-overlapping cellular targets, ExoT performs several distinct virulence functions for P. aeruginosa, including induction of apoptosis in a variety of target host cells. Both the ADPRT and the GAP domain activities contribute to ExoT-induced apoptosis. The ADPRT domain of ExoT induces atypical anoikis by transforming an innocuous cellular protein, Crk, into a cytotoxin, which interferes with integrin survival signaling. However, the mechanism underlying the GAP-induced apoptosis remains unknown. In this study, we demonstrate that the GAP domain activity is both necessary and sufficient to induce mitochondrial (intrinsic) apoptosis. We show that intoxication with GAP domain results in: (i) JNK1/2 activation; (ii) substantial increases in the mitochondrial levels of activated pro-apoptotic proteins Bax and Bid, and to a lesser extent Bim; (iii) loss of mitochondrial membrane potential and cytochrome c release; and (iv) activation of initiator caspase-9 and executioner caspase-3. Further, GAP-induced apoptosis is partially mediated by JNK1/2, but it is completely dependent on caspase-9 activity. Together, the ADPRT and the GAP domains make ExoT into a highly versatile and potent cytotoxin, capable of inducing multiple forms of apoptosis in target host cells.     

Proapoptotic Activities of Protein Disulfide Isomerase (PDI) and PDIA3 Protein,a Role of the Bcl-2 Protein Bak

Protein disulfide isomerase (PDI) family proteins are classified as enzymatic chaperones for reconstructing misfolded proteins. Previous studies have shown that several PDI members possess potential proapoptotic functions. However, the detailed molecular mechanisms of PDI-mediated apoptosis are not completely known. In this study, we investigated how two members of PDI family, PDI and PDIA3, modulate apoptotic signaling. Inhibiting PDI and PDIA3 activities pharmacologically alleviates apoptosis induced by various apoptotic stimuli. Although a decrease of PDIA3 expression alleviates apoptotic responses, overexpression of PDIA3 exacerbates apoptotic signaling. Importantly, Bak, but not Bax, is essential for PDIA3-induced proapoptotic signaling. Furthermore, both purified PDI and PDIA3 proteins induce Bak-dependent, but not Bax-dependent, mitochondrial outer membrane permeabilization in vitro, probably through triggering Bak oligomerization on mitochondria. Our results suggest that both of PDI and PDIA3 possess Bak-dependent proapoptotic function through inducing mitochondrial outer membrane permeabilization, which provides a new mechanism linking ER chaperone proteins and apoptotic signaling.     

凋亡相关基因Bcl-2、Bax及细胞色素C在糖尿病胃组织壁细胞中的表达

目的观察糖尿病大鼠胃组织中Bcl-2、Bax及细胞色素C表达变化。方法SD雄性大鼠腹腔注射链脲佐菌素(STZ)复制糖尿病动物模型,分别于4周、12周后测体重和血糖,胃内色素排空检测,光镜观察胃组织形态学变化,免疫组织化学观察Bcl-2、Bax及细胞色素C蛋白表达变化。结果糖尿病组大鼠胃内色素残留率显著增加,胃壁细胞胞质空泡改变。与正常对照组比较糖尿病组大鼠胃壁细胞Bax及细胞色素C表达随病程延长显著增加,而Bcl-2随病程延长表达减弱。结论Bcl-2、Bax及细胞色素C在糖尿病胃组织壁细胞中出现了一定的变化规律。引起壁细胞凋亡增加,导致胃粘膜功能异常,这可能是糖尿病胃瘫的重要发病机制之一。     

瑞舒伐他汀对兔心肌梗死后心室重构及细胞凋亡的影响

目的:观察瑞舒伐他汀对新西兰大白兔心肌梗死后左室重构及心肌细胞凋亡的影响。方法:45只雄性新西兰大白兔随机分成三组,即:假手术组(S,n=15),心肌梗死对照组(MI,n=15),心肌梗死瑞舒伐他汀干预组(R,n=15)MI组和R组大鼠结扎左冠状动脉前降支建立AMI模型。心肌梗死对照组及假手术组术后24h灌等量的生理盐水。瑞舒伐他汀干预组于术后24h直接灌胃法给药,给药剂量以10mg/kg*d计算。干预2周后,进行心脏超声测定,随即处死新西兰大白兔、取出心脏,观察病理组织形态,并通过Western blot方法检测Bcl-2,Bax在心肌梗死边缘区的蛋白表达。结果:①心脏超声检测表明干预组左室舒张末内径(LVEDD)、左室收缩末内径(LVESD)较心肌梗死对照组降低而左室射血分数(LVEF)较心肌梗死对照组增高②干预组心肌梗死边缘区Bax表达与心肌梗死对照组比较未见统计学差异,而Bcl-2表达高于心肌梗死对照组。结论:瑞舒伐他汀上调Bcl-2的表达,改善心室重构