[Establishment of a high throughput screening model of cannabinoid receptor 2 agonist].

AIM: To establish a drug screening model of CB(2) agonist in vitro based on signal regulation pathway. METHODS: Plasmid pIRES(2)-EGFP-CB(2), pGL(4), 29[luc2P/CRE/Hygro] and PRL-TK were co-transfected into CHO cells in 96 wells plate, to screen agonists of CB(2) receptor by detecting the expressing levels of dual luciferase activity. The concentration and acting time of the agonist were optimized and the stability of the model were investigated. RESULTS: The largest relative induction activity was obtained after 8h drug administration. Establishment of a high throughput screening model for CB(2) receptor agonist. The Z' factor is 0.75 demonstrating its perfect stability. CONCLUSION: Successfully establish a drug screening model of CB(2) agonist, which provided a basis for searching valid material from traditional Chinese medicine.

[Effect of different intensities of electroacupuncture on expression of monocyte chemoattractant protein-1 and TNF-alpha in adipose tissue in obesity rats].

OBJECTIVE: To observe the effect of different intensities of electroacupuncture (EA) on adipose tissue inflammatory cytokines in rats with simple obesity so as to investigate its mechanism underlying body weight reduction. METHODS: Forty SD male rats were randomly divided into normal, model, strong EA and weak EA groups (n = 10/group). Obesity model was established by feeding the rats with high fat diet. EA (20 Hz, strong EA: 5 V, weak EA: 2. 5 V) was applied to "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) for 15 min, once everyday and for 14 days. Body weight and Lee's index (= body weight(1/3) x 10(3) / body length) were detected. The fasting blood glucose was detected by hexokinase method, serum triglyceride (TG) was detected by glycerol-phosphoric acid oxidase peroxydase (GPO-POD)method, total cholesterol (TC) was detected by cholesterol oxidase method, high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-O) were measured by using one-step method, respectively. The expression of monocyte chemoattractant protein-1 (MOP-1) mRNA and tumor necrosis factor (TNF)-alpha mRNA in the epididymis adipose tissue was detected by reverse transcription-polymerase chain reaction (RT-POR). RESULTS: In comparison with the normal group, the body weight, Lee's index, blood lipid (TG, TC, LDL-C), fasting blood glucose levels, and expression of MOP-1 mRNA and TNF-alpha mRNA were significantly higher in the model group (P < 0.01), and HDL-C was significantly lower (P < 0.01). After EA,compared with the model group, the body weight, Lee's index, TG, TC, LDL-C, fasting blood glucose levels, and expression of MCP-1 mRNA and TNF-a mRNA in both strong and weak EA groups were significantly decreased (P < 0.01, P < 0.05), and HDL-C was significantly increased (P < 0.01, P < 0.05). The effects of strong EA group were obviously superior to those of weak EA group (P < 0.05, P < 0.01). No statistical significance was observed between the two EA groups in fasting blood glucose levels (P > 0.05). CONCLUSION: EA of "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) has a beneficial weight-reduction effect on rats with simple obesity, and moreover, the effect of strong EA stimulation is evidently superior to weak EA stimulation.

Effect of Shenmai injection on expression and activity of heme oxygenase-1 in reperfusion injury after pulmonary ischemia in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effect of Shenmai injection the expression of heme oxygenase-1 (HO-1) in rabbits with reperfusion injury after pulmonary ischemia.</p><p><b>METHOD</b>Single lung ischemia/reperfusion injury animal model was used in vivo. Twenty rabbits were randomly divided into two groups (n = 10, in each), pulmonary ischemia and reperfusion injury (PIRI) group and I-R + Shenmai injection group. The tissue slides were stained by in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the absorbance. Wet to dry ratio of lung tissue weight (W/D) and the injured alveoli rate (IAR) were measured at 180 minutes after lung reperfusion. Meanwhile the lung tissue slide was prepared for electron microscopic observation at 180 minutes after reperfusion.</p><p><b>RESULT</b>HO-1 expression was upregulated in two groups in the pulmonary endothelial cells, part of pulmonary vascular smooth muscle cells, extima of vessels and epithelial cells of airway, the absorbance was 0.148 +/- 0.013, 0.158 +/- 0.012, respectively. The Shenmai injection group showed higher absorbance than those of the IRI group (P < 0.01), lower W/D and IAR values than those of the IRI group (P < 0.01) significantly and lighter abnormal changes of the lung tissue in morphologically than those of the PIRI group.</p><p><b>CONCLUSION</b>Shenmai injection possesses notable protective effects on PIRI in rabbits by increasing the expression of HO-1 in lung.</p>

Metabolic adaptations in skeletal muscle during lactation: complementary deoxyribonucleic acid microarray and real-time polymerase chain reaction analysis of gene expression.

Lactation and fasting are two physiological models characterized by negative energy balance. Our previous studies demonstrated that uncoupling protein (UCP) 3 expression in skeletal muscle was down-regulated during lactation and up-regulated during fasting. The present studies used cDNA microarray and real-time PCR to perform a systems and comparative analysis in gene expression in skeletal muscle under conditions of negative energy balance. Gastrocnemius skeletal muscle RNA pools were generated from the following groups of rats: cycling diestrous females, cycling females with 48 h of fasting, lactation, and lactation + leptin. Of those known genes studied, 35 genes were up-regulated and 49 were down-regulated during lactation. Leptin treatment during lactation reversed the differential regulation of about 80% of these genes, demonstrating the importance of the leptin suppression to the changes in skeletal muscle metabolism. GenMAPP analysis revealed a coordinated regulation at key steps in glycolysis/gluconeogenesis, the tricarboxylic acid cycle, and lipid metabolism, indicating an increased rate of lactate production through glycolysis and reduced fatty acid degradation in skeletal muscle during lactation. Particular interest was paid to those genes that changed in a similar manner to UCP3 mRNA. Many of these genes that were decreased during lactation and increased during fasting are involved in fatty acid degradation and transport, including acyl-coenzyme A dehydrogenase for medium chain fatty acid, carnitine palmitoyltransferase 1, and fatty acid translocase. The current studies provide a basis for investigating the mechanisms underlying metabolic adaptations during lactation and fasting and highlight the importance of UCP3 in lipid metabolism.