Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice.@*METHODS@#siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR.@*RESULTS@#Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01).@*CONCLUSIONS@#The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

Anchusa italica extract: phytochemical and neuroprotective evaluation on global cerebral ischemia and reperfusion

Abstract Stroke is the third leading cause of mortality and disability in industrial countries. Treatment with herbs with antioxidant properties has been reported to be an alternative to the conventional treatments. This study was conducted to investigate the effect of Anchusa italica extract on hippocampal injury induced by transient global cerebral ischemia and reperfusion in the rat. To do so, 50 rats were randomly assigned to five groups; control, sham, ischemia, and 50 or 100 mg/kg of Anchusa italica treated animals. Ischemia was induced by occlusion of carotid artery for 30 minutes. Afterward, behavioral tests and biochemical analyses were conducted. Induction of ischemia/reperfusion caused a decline in learning and passive avoidance memory in rats. Moreover, Anchusa italica caused an increase in learning and improved the passive avoidance memory. Induction of ischemia/reperfusion caused a decrease in the antioxidant capacity of the brain and serum as well as an increase in the malondialdehyde of the brain and serum. Anchusa italica led to an increase in the antioxidant capacity of the brain and serum and decrease in the malondialdehyde of the brain and serum. Overall, because of its protective effects on spatial memory, passive avoidance learning, antioxidant capacity, and lipid peroxidation during ischemia/reperfusion, Anchusa italica might be beneficial in ischemic patients.

Antitumor function and mechanism of phycoerythrin from Porphyra haitanensis

The anti-tumor effect of R-Phycoerythrin (R-PE) from Porphyra haitanensis was studied using cell line HeLa as an in vitro model and Sarcoma-180 (S180) tumor-bearing mice as an in vivo model. The results showed that the combination treatment of R-PE and photodynamic therapy PDT) significantly inhibited the growth of HeLa cells up to 81.5%, with a fair dose-effect relationship, but did not inhibit endothelial cells. The annexin v-fitc/PI fluorescence staining experiments demonstrated that at doses between 0~60µg/mL, apoptosis cells and later stage apoptosis cells or necrosis cells increased significantly as the R-PE dosage increased. DNA electrophoresis showed that after R-PE+PDT treatment of HeLa cells for 24 hours, a light "smear" band between 100~400bp appeared to indicate the degradation of genomic DNA. The QRT-PCR results showed that R-PE+PDT treatment increased caspase-3 and caspase-10 gene expression and decreased the Bcl-2 gene expression level significantly as the R-PE dose increased, implying that R-PE promoted HeLa cell apoptosis. Compared with untreated S180 tumor-bearing mice, R-PE injection significantly inhibited the growth of S180 in tumor-bearing mice up to 41.3% at a dose of 300mg-kg-1. Simultaneously, the significant increase of superoxide dismutase (SOD) activity in serum (p < 0.01) and the decrease of the malondialdehyde (MDA) level in liver suggests that R-PE improved the anti-oxidant ability of the S180 tumor-bearing mice, which may related to its antitumor effect. In addition, the R-PE caused a significant increase (p < 0.05) in the spleen index and thymus index, and a significant increase (p < 0.01) in lymphocyte proliferation, NK cell kill activity and the TNF-α level in the serum of S180 tumor-bearing mice. These results strongly suggest that the antitumor effect of R-PE from Porphyra haitanensis functioned by increasing the immunity and antioxidant ability of S180 tumor-bearing mice, promoting apoptosis by increasing protease gene expression and TNF-α secretion.

Effects of ethyl pyruvate on myocardial apoptosis and expression of Bcl-2 and Bax proteins after ischemia-reperfusion in rats / 华中科技大学学报（医学）（英德文版）

In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion (I/R) in vitro and the expression of Bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups (n=8 in each group): control group was perfused for 120 min. In the I/R group, after 30 min stabilization the injury was induced by 30 min global ischemia followed by 60 min reperfusion. Ethyl pyruvate (EP) group was set up with the same protocol as I/R group except that it was supplied with 2 mmol/L EP 15 min before ischemia and throughout reperfusion. Myocardial malonaldehyde (MDA) content was measured. Myocardial apoptotic index (AI) was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. The expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of Bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of Bcl-2 protein was up-regulated in EP group (P<0.05). These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating Bcl-2 and down-regulating Bax proteins.

Effect of amaranth leaves on dimethylhydrazine-induced changes in multicomponent antioxidant system of rat liver.

Effect of prefeeding dehydrated amaranth (A. gangeticus) leaves at 10 and 20% levels on a chemical toxicant, dimethylhydrazine (DMH)-induced free radical stress in rat liver was evaluated. DMH-induced rise in hepatic malondialdehyde (MDA), was diminished by AL. AL intake resulted in a significant increase in hepatic glutathione (GSH). The feeding of AL at 10% level increased the hepatic glucose-6-phosphate dehydrogenase (G-6-PDH) activity, while that at 20% level increased the hepatic glutathione reductase (GSSGR) as well, in addition to G-6-PDH. Amaranth leaves at 10 and 20% levels of feeding diminished the hepatic superoxide dismutase and glutathione peroxidase (GSH-Px) activities. DMH influenced adversely the hepatic antioxidant enzyme activities. Simultaneous administration of DMH and feeding of AL enhanced the DMH-induced decrease in hepatic GSH-Px. DMH enhanced formation of micronuclei was reverted significantly by AL intake. Hence, it was concluded that the consumption of AL at 20% level reduced DMH-induced impaired antioxidant status in rat liver.

Autoantibody against, Malondialdehyde-Modified Low Density Lipoprotein in Patients with Non-Diabetic Unstable Angina: A Potential Role in Immunologic Reaction of Plaque Instability

The role of autoantibody against oxidized low-density lipoprotein (LDL) in the pathogenesis of atherosclerosis is still unknown. The purpose of this study was to determine whether autoantibodies against malondialdehyde (MDA)-modified LDL are associated with coronary artery disease (CAD) and clinical presentations of CAD in non-diabetic patients without acute myocardial infarction (AMI). We determined the serum levels of autoantibody against MDA-modified LDL by ELISA in 71 patients with angiographically significant CAD (> or = 50% diameter stenosis in at least 1 vessel) and 80 controls without angiographically significant CAD. Among the total 151 subjects, 30 subjects did not have any clinical ischemic event, 90 subjects had stable angina symptoms, and 31 subjects had unstable angina symptoms. The autoantibody titer, expressed mean optical density units, was significantly higher in patients with CAD than in controls (0.177+/- 0.014 versus 0.127+/- 0.011, respectively; p=0.006) and higher in unstable angina group than in stable angina group (0.240+/- 0.025 versus 0.145+/- 0.007, respectively; p < 0.001). By logistic regression analysis, the high autoantibody titer was associated significantly with CAD (P=0.008), independent of age, gender, body mass index, triglyceride concentration, and the ratio of total cholesterol-high density lipoprotein (HDL) cholesterol. In multiple regression analysis, presence of CAD, smoking history and low HDL-cholesterol level were independent factors associated with a increased anti-oxLDL Ab titer. The autoantibody titer was significantly lower in nonsmoker than smoker (p=0.019) and higher in low HDL- cholesterol (< or = 35 mg/dl) group than in high HDL-cholesterol group (p=0.012). Elevated autoantibody titer was associated with CAD and the unstable clinical presentation of CAD. Our results suggest that immune response to oxidized LDL may play a role in the pathogenesis of atherosclerosis and plaque instability.

Effect of melatonin on analgesic and anti - inflammatory action of indomethacin in rats with induced adjuvant arthritis

This experimental study was carried out to evaluate the possible therapeutic effects of melatonin in rheumatoid arthritis and its interaction with indomethacin. The study was carried out on rats in which rheumatoid-like arthritis were induced experimentally by an intradermal injection of a total volume of 0.1 ml of cold emulsion consisting of native type II collagen and complete Freund's adjuvant. A booster dose was given after three weeks. It could be concluded that melatonin had an immune stimulant effect enhancing the immune system and its use in autoimmune diseases such as rheumatoid arthritis exaggerated arthritic manifestations. Also, a combined administration of melatonin and indomethacin had no therapeutic effect on patients suffering from rheumatoid arthritis

Estudo da peroxidaçäo lipídica em ratos deficientes em vitamina E submetidos à nefrectomia subtotal / Study vitamin E deficiency on lipid peroxidation in rats submitted to subtotal nefrectomy

Evidências experimentais e clínicas têm indicado que as manifestaçöes da Insuficiência Renal Crônica (IRC) estäo relacionadas com um aumento na produçäo dos radicais livres. Pacientes portadores de IRC apresentam aumento da peroxidaçäo lipídica com conseqüente diminuiçäo progressiva da vitamina E, um dos mais importantes antioxidantes. Neste estudo foi avaliado o papel da deficiência da vitamina E sobre a peroxidaçäo lipídica em ratos submetidos à nefrectomia subtotal. O estudo incluiu ratos macho, Wistar, mantidos sob dieta durante 45 dias, de acordo com os seguintes Grupos: GSDN - sham dieta normal; GNDN - nefrectomizado dieta normal; GSDD - sham dieta deficiente; GNDD - nefrectomizado dieta deficiente. Após 30 dias de estudo, os animais Experimentais foram submetidos à 5/6 de nefrectomia, enquanto os ratos Controle ao sham operatório...

Níveles plasmáticos de un marcador de peroxidación lipídica y de vitamina E en mujeres sanas eumenorreicas, en postmenopáusicas sanas y en primigestas que desarrollaron o no hipertensión inducida / Even plasmatic od a marker of lipid peroxidation and E vitamin in women healthy eumenorreicas, in health postmenopausal and in primigravidas that develope or not induced hypertension

Modificaciones en níveles circulantes de malondialdehido y de vitamina E se han asociaado con hipertensión inducida por embarazo, con diabetes gestacional y con parto pretérmino. Hay solo información fragmentaria aa esstos respectos

Efectos de sulfinpirazona sobre tejidos miocárdicos con estimulación eléctrica o sin ella / Effects of sulfinpyrazone on myocardial tissues with or without electrical stimulation

Se estudiaron in vitro los efectos del ácido arquidónico (AAq) y sulfinpirazona (SP) sobre miocardio de rata "suquía" de 240+ ou - 10g. Bandas auriculares y ventriculares fueron instaladas en solución Krebs-bicarbonato a 36 + ou- 1-C, registrado tensión contráctil isométrica (TC) y niveles de malonato (MDA), con electroestimulación o sin ella (1,6 Hz), AAq (2,5 microng/ml) y SP (375, 750, 1.500 y 3.000 microng/ml). AAq deprimió 100% de la TC de ambos preparados, registrándose en tiempos variables (9 a 17 min.) recuperación espontánea en la mayoría de los casos, aunque sin recuperar los valores iniciales. La reactividad eléctrica aumentó en el tejido auricular, pero no en el ventricular. Los efectos de SP dependen de la concentración; 375 microng/ml bajan la TC auricular 87 + ou - 7,3%. La respuesta ventricular a esa dosis fue variable, aunque la TC disminuyó en 61,5% de los preparados; 3 mg/ml son homogéneamente estimulantes. La reactividad eléctrica disminuye con SP. La concentración de MDA en los preparados auriculares cayó muy significativamente (p<0,001) después de SP. En los ventriculares se observó disminución de menor magnitud, también significativa (p<0,02). La suma algebraica de caída y recuperación de TC de los preparados auriculares después de dar SP, da un porcentaje de pérdida similar al descenso del malonato (+ ou - 25%). Se sugiere que: a) AAq deprime el miocardio por acción directa sin descartar alguna transformación inmediata en derivados activos. b) La SP inhibiría distintas enzimas en función de las dosis. c) El MDA parece ser consumido por el tejido miocárdico o metabolizado en el medio. Se plantean explicaciones alternativas para este fenómeno y su relación con el mecanismo de acción de la sulfinpirazona