Effects of Genipin on the expression of uncoupling protein 1 in brown adipose and white adipose tissues in mice / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of genipin on promoting brown adipose tissue activation and white adipose tissue browning.@*METHODS@#The male C57BL/6J mice were divided into three groups: normal control group, genipin group and cold-stimulus group.Genipin group were treated consecutively with genipin at a dose of 15 mg/kg once a day for 9 days, normal control group were treated with the saline.The mice with cold-stimulus were exposed to 4℃ environment for 5 days.Daily food amount and body weight were measured.Morphological changes were observed in the subscapular region, inguinal region and epididymis around the adipose tissue.The expression of uncoupling protein 1 (UCP1) was determined by real-time PCR and Western blot respectively.@*RESULTS@#The wet weight of white fat in genipin-treated mice was decreased by 16% , and 28% in that of cold-stimulus mice, compared with the normal control group (P＜0.05).After treatments of genipin and cold-stimulus, the color of white adipose tissues was darker, and the size of lipid droplets in adipocytes was smaller, whereas the number was increased.Compared with the normal control group, UCP1 expression was increased obviously in fat tissues, including the subcutaneous and visceral white adipose tissues, and brown adipose tissue after treated with genipin and cold-stimulus (P＜0.05).@*CONCLUSION@#Genipin promoted activation of brown adipose tissue and browning of white adipose tissue by upregulating UCP1 expression, which could contribute to the loss of body weight against obesity.

Establishment of the Chang liver cell line stably overexpressing human UCP2 gene and its effect on mitochondrial membrane potential and reactive oxygen species / 中华肝脏病杂志

To establish the Chang liver cell line stably overexpressing human uncoupling protein 2 (UCP2) and observe the effect of UCP2 on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The Chang liver cell line was transfected with recombinant plasmid containing full-length human UCP2 cDNA (pcDNA3.1-hUCP2) or pcDNA3.1 empty vector. The stable cell line was established by antibiotic screening with Zeocin. UCP2 expression was detected by Western blotting and immunocytochemistry. The UCP2 overexpressing cells were pretreated with genipin at various doses (25, 50 and 100 munol/L). MMP and intracellular ROS were detected by fluorescence spectrophotometry. The total normalized protein content in UCP2 overexpressing cells was 1.6-fold higher than that in unmanipulated normal cells. The fluorescence intensities of Rhodamine123 and DCFH-DA in UCP2 overexpressing Chang liver cells (11.11+/-2.76 and 4.97+/-0.62, respectively) were significantly lower than those in unmanipulated normal cells (15.56+/-2.55, P less than 0.01 and 6.14+/-1.25, P less than 0.05, respectively) and in cells transfected with empty vector (16.11+/-2.93, P less than 0.01 and 6.23+/-1.13, P less than 0.05, respectively). Treatment of UCP2 overexpressing cells with 25, 50 and 100 munol/L genipin caused a dose-dependent increase in fluorescence intensities of Rhodamine123 (14.89+/-2.89, 17.89+/-2.93 and 24.00+/-2.55, respectively, all P less than 0.01) and DCFH-DA (9.16+/-0.78, 10.84+/-1.09 and 11.83+/-1.25, respectively, all P less than 0.01). The Chang liver cell line stably overexpressing UCP2 was established successfully. Using this cell system, UCP2 was found to play a role in mitochondrial function by regulating MMP and ROS.

Establishing the model of kappaB-decoy inhibiting the activity of NF-kappaB in PC12 cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>Observing the time course and establishing the model of kappaB-decoy oligodeoxynucleotides (rcB-decoy) inhibiting the activity of NF-kappaB in the PC12 cells.</p><p><b>METHODS</b>PC12 cells cultivating in the 6 wells plate were divided into 3 groups, experimental group: adding kappaB-decoy complex (6 microg DNA/well), the control group: adding scrambled-decoy complex, the normal group: adding lipid-Lipofectamine 2000, transfer and cultivate 48 h, then lipopolysaccharide (LPS, 200 ng/ml) was added in the cells for 0.5-4 h. The immunocytochemistry and Western blot were used to measure the expression or the activity of NF-kappaB in PC12 cells.</p><p><b>RESULTS</b>In PC12 cells, compared with normal group, the expression of NF-kappaB enhanced obviously with the time of the stimulation of LPS in scrambled-decoy treated control group (P < 0.01), in 2-4 h the level reached the peak; the expression of NF-kappaB showed the stable level with the time of the stimulation of LPS in kappaB-decoy treated experimental group, compared with the control group, the expression levels were obviously lower than the respective time point of control groups (P < 0.01).</p><p><b>CONCLUSION</b>kappaB-decoy could reduce the expression of NF-kappaB in the normal PC12 cells and inhibit the activity of NF-cB in the pathologic PC12 cells.</p>

Protective effects of glucagon-like peptide 2 on intestinal ischemia/reperfusion injury in mice / 中国应用生理学杂志

<p><b>AIM</b>To investigate the protective effects of glucagon-like peptide 2(GLP-2) on intestinal ischemia/reperfusion (I/R) injury in mice.</p><p><b>METHODS</b>Intestinal ischemia/reperfusion model in mice were set up and 32 mice of Kunming species were divided randomly into 4 groups (n=8): Sham group, I/R group, I/R + GLP-2 group and I/R + glutamine group. The morphologic changes of intestinal mucosa were observed under LM. The villus height and crypt depth of intestine, the activity of diamine oxidase (DAO) in intestine and bacterial translocation rates of mesenteric lymph nodes (MLN) were detected.</p><p><b>RESULTS</b>Compared with sham operation group, the intestinal villi were sloughed in I/R group with decreased villus height and crypt depth (P < 0.01), the DAO activities were decreased (P < 0.01), and MLN bacterial translocation rates were increased (P < 0.05). While GLP-2 administration improved the villus damage, increased DAO activity (P < 0.01), and decreased MLN bacterial translocation rates (P < 0.05), compared with I/R group.</p><p><b>CONCLUSION</b>GLP-2 have protective effects on intestinal morphology and barrier function after ischemia/reperfusion injury in mice.</p>

Effects of vagotomy on UCP2 mRNA expression and gastric acid secretion in rats / 中国应用生理学杂志

<p><b>AIM</b>The relationship between gastric acid secretion and ATP level, and regulation of uncoupling protein 2 (UCP2) mRNA expression by vagus nerve were studied in vagotomies rats.</p><p><b>METHODS</b>With the high selective vagotomy model, the gastric acidity was titrated to pH 7.0 with 0.1 mol/L NaOH solution and ATP contents were quantified by using fluorimetry. The expression of UCP2 mRNA was observed by using Northern blot in stomach of rats.</p><p><b>RESULTS</b>Both of gastric acidity and ATP contents in stomach body decreased significantly at 24 h after vagotomy. The expression of UCP2 mRNA was markedly increased as compared with sham operation group.</p><p><b>CONCLUSION</b>ATP contents decreased and vagus nerve down-regulates expression of UCP2 mRNA in stomach corpus in vagotomies rats. The results indicates that vagus nerve could underlay the gastric acidity by inhibiting expression of UCP2 mRNA and increasing ATP contents in rats.</p>

Effect of no mediator on kainic acid induced behavioral seizures in rats / 中国应用生理学杂志

<p><b>AIM</b>To further explore the roles of endogenous nitric oxide (NO) or NO derivatives in complex partial seizures and generalized convulsions.</p><p><b>METHODS</b>The effect of pretreatment with L-nitroarginine (L-NNA), an inhibitor of nitric oxide synthase (NOS), or L arginine (L-Arg), a precursor of NO on kainic acid (KA)-induced seizure in rats and the changes in the concentration of NO2 -/NO- in the hippocampus were determined.</p><p><b>RESULTS</b>The rats appeared with wet dog shakes (WDS) at 15 min and then occurred generalized convulsions during 1 h to 3 h after administration of KA (10 mg/kg i.p.). However, the pretreatment of L-NNA (50 mg/kg) so dramatically promoted and enhanced KA-induced behavioral seizures that the latency of generalized convulsion was shorten dramatically, and the mortality was greatly high. In contrast, the pretreatment with L-Arg (40 mg/kg) markedly delayed or weakened KA-induced behavioral changes, such as increasing latency of WDS and generalized convulsion, shortening time o f seizure and none of animal died during observed time. The concentration of NO2- /NO3- in the hippocampus increased immediately at 30 min and remained to 7 d after the administration of KA. Compared with control group (pretreatment with NS), the concentration of NO2- / NO3- in the hippocampus apparently increased at 3 h and 3 d after the administration of KA in the rats with L-Arg pretreatment.</p><p><b>CONCLUSION</b>The endogenous NO (NO or NO derivatives) mediators may play an important role against excitotoxin induced seizures in rats.</p>