Objective To investigate the effects of N - acetylcysteine (NAC) on apoptosis and the expressions of Fas/FasL mRNA in lung tissue of rats with paraquat - induced acute lung injury.Methods Forty five male SD rats were randomly (random number) divided into normal control group,paraquat (PQ) group,and NAC treatment group.The rat model of acute lung injury was made with 2％ PQ induction in dose of 25 mg/kg injected,and NAC was injected into the PQ poisoning rats (200 mg/kg) 30 minutes after PQ administration in NAC treatment group.In the control group,equal amount of saline instead was injected into the rats.Apoptosis was detected by using TUNEL method and the expressions of Fas/FasL mRNA were evaluated by using reverse transcription polymerase chain reaction (RT- PCR),and the levels of Fas/FasL protein were detected by using western blot analysis.Results Compared with control group,cell apoptosis and expressions Fas/FasL mRNA in PQ group were significantly different ( P ＜ 0.05 ).Compared with PQ group,cell apoptosis and expressions Fas/FasL mRNA in NAC group were significantly decreased,were significant lower (P ＜ 0.05).Conclusions NAC inhibited apoptosis in lung tissue of rats with paraquat induction by regulating the activation of Fas/FasL systems.

Objective To study the mechanism of paraquat (PQ)-induced renal injury in rats,the expression changes of ICAM-1 to assess the protective effect of Melatonin in PQ poisoning.Methods Ninety adult healthy Sprague-Dawley (SD) rats were divided into three groups at random.Control group:30 rats;Poisoned group:30 rats;Melatonin group:30 rats.Control group and Poisoned group were treated intragastrically with 1 ml of PQ (50 mg/kg) diluted with normal saline.Control groupwere treated with the same dose of normal saline as Poisoned group and Melatonin group.Melatonin group were given 1 ml of Melatonin at a dose of 10 mg/kg diluted with normal saline (once daily,intraperitoneally) Control and Poisoned group were treated with the same dose of normal saline (once daily,intraperitoneally) as Melatonin group.Pathology of renal tissue were oberserved by HE staining,and electron microscope.The histopathological changes and the expression of ICAM-1 were observed with mmunohistochemistry (IHC).Results (1) There were no obvious pathological changes in Control group.Poisoned group Renal glomerulus had hyperemia and distension.Renal tubule epithelial cell had edema and vacuolar degeneration and renal tubule lumina was narrowing on day 1,There were serious edema exudation and necrosis on day 5,which gradually lessened furthermore;Compared with Poisoned group,the aforementioned pathological lesion was more palliative in Melatonin group.(2) No obvious abnomal changes in ultrastructure of renal tissues in Control group.There were swelling of mitochondrion and rupture of renal tubule epithelial cell and endoplasmic reticulum had extension,lysosome was mult and had much phagocytosis in Poisoned group.(3)There was a very weak expression of ICAM-1 in Control group.while in Poisoned group,there was already a significant higher expression of ICAM-1 of renal tubule on day 1 after PQ poisoning,Immunohistochemistry score (IHS) of Poisoned group on day 1,3,5,7,14 were (0.1561 ±0.0295、0.2572±0.0259、0.3028±0.0153、0.2083 ±0.0227、0.9309 ±0.0059),compared with Control group (P＜0.01);Melatonin group were (0.1259±0.0061、0.2109±0.0280、0.2679±0.0233、0.1771 ±0.0186、0.0791 ±0.0135),compared with Control group (P＜0.01),compared with Poisoned group (P＜0.05);Conclusion ICAM-1 was involved in the procedures of renal injury;MT surely had a protective effect,which might be mediated by ICAM-1 in the paraquat-induced renal injury,but its regulation path still need a further exploration.

Objective To study the mechanism of paraquat (PQ)-induced renal injury in rats,the expression changes of ICAM-1 to assess the protective effect of Melatonin in PQ poisoning.Methods Ninety adult healthy Sprague-Dawley (SD) rats were divided into three groups at random.Control group:30 rats;Poisoned group:30 rats;Melatonin group:30 rats.Control group and Poisoned group were treated intragastrically with 1 ml of PQ (50 mg/kg) diluted with normal saline.Control groupwere treated with the same dose of normal saline as Poisoned group and Melatonin group.Melatonin group were given 1 ml of Melatonin at a dose of 10 mg/kg diluted with normal saline (once daily,intraperitoneally) Control and Poisoned group were treated with the same dose of normal saline (once daily,intraperitoneally) as Melatonin group.Pathology of renal tissue were oberserved by HE staining,and electron microscope.The histopathological changes and the expression of ICAM-1 were observed with mmunohistochemistry (IHC).Results (1) There were no obvious pathological changes in Control group.Poisoned group Renal glomerulus had hyperemia and distension.Renal tubule epithelial cell had edema and vacuolar degeneration and renal tubule lumina was narrowing on day 1,There were serious edema exudation and necrosis on day 5,which gradually lessened furthermore;Compared with Poisoned group,the aforementioned pathological lesion was more palliative in Melatonin group.(2) No obvious abnomal changes in ultrastructure of renal tissues in Control group.There were swelling of mitochondrion and rupture of renal tubule epithelial cell and endoplasmic reticulum had extension,lysosome was mult and had much phagocytosis in Poisoned group.(3)There was a very weak expression of ICAM-1 in Control group.while in Poisoned group,there was already a significant higher expression of ICAM-1 of renal tubule on day 1 after PQ poisoning,Immunohistochemistry score (IHS) of Poisoned group on day 1,3,5,7,14 were (0.1561 ±0.0295、0.2572±0.0259、0.3028±0.0153、0.2083 ±0.0227、0.9309 ±0.0059),compared with Control group (P＜0.01);Melatonin group were (0.1259±0.0061、0.2109±0.0280、0.2679±0.0233、0.1771 ±0.0186、0.0791 ±0.0135),compared with Control group (P＜0.01),compared with Poisoned group (P＜0.05);Conclusion ICAM-1 was involved in the procedures of renal injury;MT surely had a protective effect,which might be mediated by ICAM-1 in the paraquat-induced renal injury,but its regulation path still need a further exploration.

The aim of the study was to construct a recombinant adenovirus overexpression vector for Sterol Regulatory Element Binding Protein-1 (SREBP-1) of Xinong Saanen dairy goat, and to detect its effect on genes related to fatty acid metabolism in goat mammary epithelial cells, to establish foundation for further study of its roles in metabolism of fatty acid synthesis and lactation. First, we designed primers based on the SREBP-1 gene sequence in GenBank for PCR amplification and inserted the sequence into shuttle vector pAdTrack-CMV. The recombinant plasmid pAdTrack-CMV-SREBP-1 linearized by Pme I was transformed into E. coli BJ5183 competence cell containing the backbone vector pAdEasy-1 to obtain recombinant vector pAd-SREBP-1 by homologous recombination. pAd-SREBP-1 was linearized by Pac I and transfected into HEK 293 cell. Then we infected goat mammary epithelial cells with recombinant adenovirus which was packaged in HEK 293 cell line. The results showed that the recombinant adenovirus vector containing SREBP-1 was successfully constructed, and the titer of virus was 10(9) U/mL. Compared with the control group, mRNA level of SREBP-1 increased by about 15 times after infected for 48 h and 30 times after infected for 72 h. Fatty acid synthase (FASN) and Acetyl-CoA carboxylase (ACC) was upregulated by almost 2 times. The expression level of Peroxisome proliferator activated receptorgamma (PPARgamma) increased by 1.5 times. Liver X receptoralpha (LXRalpha) and Adipose triglyceride lipase (ATGL) upregulated by 1.2 times compared with that of control. But Stearoyl-coenzyme A desaturase (SCD) had no obvious change. In conclusion, SREBP-1 can activate the expression of genes related to fatty acid synthesis in mammary epithelial cells of Xinong Saanen dairy goat, demonstrated a regulatory function on the fatty acid metabolism in goat mammary gland.
Adenoviridae ; genetics ; metabolism ; Animals ; Epithelial Cells ; cytology ; metabolism ; Fatty Acids ; metabolism ; Female ; Gene Expression Regulation ; Goats ; genetics ; HEK293 Cells ; Humans ; Lipid Metabolism ; genetics ; Mammary Glands, Animal ; cytology ; Sterol Regulatory Element Binding Protein 1 ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the effects of acetamide at different doses on the expression of inhibitory amino acids (gamma-aminobutyric acid, GABA) and excitatory amino acid (glutamate, Glu) in the cerebral cortex of rats with acute tetramine (TET) poisoning.

METHODSEighty Sprague-Dawley rats (SPF) were randomly divided into five groups, with 16 rats in each group: saline control group, dimethyl sulfoxide (DMSO) control group, TET exposure group, high-dose (2.8 g/kg/d) acetamide treatment group, and super-high-dose (5.6 g/kg/d) acetamide treatment group. Rats in the exposure group and treatment groups were exposed to TET by intragastric administration after fasting, and were then intramuscularly injected with saline or different doses of acetamide in the following 5 days. The cortex of the temporal lobe was collected at 3 h, 12 h, 48 h, or 7 d after treatment. The expression levels of GABA and Glu in the cortex of the temporal lobe were determined by average optical density (OD) values in immunohistochemistry.

RESULTS1) Expression of GABA: The OD value of GABA in TET exposure group started to increase at 12 h after treatment, reached the peak at 48 h, and decreased to the normal level at 7 d. In the high-dose acetamide treatment group, the increase in OD at 12 h was not so significant as that in the TET exposure group, OD value decreased to the normal level at 48 h and was lower than that in the exposure group, and the changes were more like those in the control groups. In the super-high-dose acetamide treatment group, OD value began to increase significantly at 3 h and was significantly higher than that in the TET exposure group (P < 0.01), it reached the peak at 12 h, and was restored to the normal value at 48 h. 2) Expression of Glu: The OD value of Glu in TET exposure group at 3 h after treatment was significantly lower than those in the two control groups, it increased gradually from 12 h to 48 h, and recovered to the normal level at the 7th d. The changes in the high-dose acetamide treatment group were similar to those in the TET exposure group, but became more like those in the control groups after 48 h; the OD value in super-high-dose acetamide treatment group was significantly higher than that in the TET exposure group at 3 h after treatment (P < 0.01), while no significant difference was found at 12 h; it was significantly lower than those of all other groups at 48 h and 7 d (P < 0.01).

CONCLUSIONSTreatment with high dose of acetamide has some curative effect on TET poisoning-induced central nervous lesion, while the effect of super-high-dose acetamide on expression of neurotransmitters is too complex to evaluate.

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; poisoning ; Cerebral Cortex ; drug effects ; metabolism ; Female ; Glutamic Acid ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVETo observe the effect of different doses of acetamide on the histopathology in the cerebral cortex of rats with tetramine (TET) poisoning and to provide a basis for the treatment of fluoroacetamide poisoning with acetamide.

METHODSEighty clean Sprague-Dawley rats were randomly divided into five groups: saline control group,dimethylsulfoxide water solution control group,TET poisoning group, acetamide (2.88 g/kg/d) treatment group, and acetamide (5.68 g/kg/d) treatment group, with 16 rats in each group. Rats in the poisoning group and treatment groups were poisoned with TET by intragastric administration after fasting; then, saline was injected intramuscularly into rats of the poisoning group, and different doses of acetamide were injected intramuscularly into rats of treatment groups; the course of treatment was 5 d. At 3 h, 12 h, 48 h, and 7 d after treatment, the cerebral cortex was harvested from rats in each group, and the histopathological changes in the cerebral cortex were evaluated under light and electron microscopes.

RESULTSThe light microscopy showed that the TET poisoning group had hypoxia changes in the cerebral cortex, which worsened over time; the treatment groups had reduced hypoxia changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group. The electron microscopy showed that the apoptosis of neuronal cells were the main pathological changes in the TET poisoning group; the treatment groups had reduced apoptotic changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group.

CONCLUSIONNo pathological changes associated with the synergistic toxic effect of acetamide and TET are found in the cerebral cortex. Acetamide (2.88 g/kg/d) could reduce central nervous lesions, but the efficacy is not improved after increasing the dose. For patients who cannot be identified with TET or fluoroacetamide poisoning, acetamide could be considered for treatment.

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; toxicity ; Cerebral Cortex ; drug effects ; pathology ; Disease Models, Animal ; Male ; Rats ; Rats, Sprague-Dawley