Laboratory Animal and Comparative Medicine

Establishment of a Standard Obliterative Bronchiolitis Rat Model for Research on MicroRNA Expression Profiles

WANG Jin-yi, CAO Hao, HONG Xuan, CHEN Guo-han, FAN Hui-min, LI Qin-chuan, LIU Zhong-min

2014, 34(3): 171-175. DOI: 10.3969/j.issn.1674-5817.2014.03.001

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Objective Exploration on simulation establishment of rat Orthotopic tracheal transplantation obliterative bronchiolitis (OB) after lung transplantation, pathophysiological process, for the study of lung transplantation in OB microRNA (miRNA) expression profiles provide an ideal animal model. Methods By Orthotopic tracheal transplantation in inbred rats, establish simulation of OB after lung transplantation animal models according to the designed groups (n=6 pairs). After successful models HE used dye to assess the tracheal transplantation model; The total RNA was extracted from transplanted trachea tissues by Trizol. Small miRNA was isolated from total RNA, hybridized on miRNA microarray, and the microarray results confirmed by quantitative real-time reverse transcription-polymerase chain reaction(RT-qPCR). Results The survival rate of established model was 86.67% (26/30) beyond four weeks. The histology suggested that the trachea endothelial cells in experiment group were damaged, accompanied by enhanced inflammatory cell infiltration and fibroproliferation. Incidence of obliterative bronchiolitis in mice of experimental group was approximately 100％. Compared with the normal control group, there were 29 specific miRNAs in the model control group with the expression difference of more than 2 times by 15 up-regulation miRNAs and 14 down-regulating miRNAs, verify which significantly increase miR-146a, miR-155, and significantly reduced miR-451 and test results with good consistency. Conclusion Stability simulation of Orthotopic tracheal transplantation OB obstructive airway disease after lung transplantation can be applied after lung transplantation animal models of miRNA expression profiles in OB research.

Comparison on Histologic Pathology in vitro of Initial Artificial Carious Lesion in Bovine and Human Enamel

CHU Jin-pu, ZHOU Xue-dong

2014, 34(3): 176-180. DOI: 10.3969/j.issn.1674-5817.2014.03.002

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Objective To pathologically evaluate the production of artificial carious lesion in bovine and human enamel in vitro and to look for teeth from other species to provide quantities of standardized material for caries disease research. Methods The Lesions in bovine and human enamel were produced by 3- day exposure to a partially saturated acidic buffer solution. The histopathology of these enamel specimens were observed and compared by microscope, polarized light microscopy and scanning electron microscope. Results All specimens in a partially saturated acidic buffer solution were observed as a white opaque spot and could morphologically form subsurface demineralization with a relatively intact surface layer ,which was the same as the early natural enamel caries, except that the lesions depth was deeper than those in human teeth. Subsurface lesions in bovine enamel and human enamel show an irregular and eroded surface with openings. The crystallites are short ,have club-like endings and irregular shape, but The crystallites in bovine enamel are less densely packed, more Larger in size than those in human enamel. Conclusion Lesions in bovine teeth were pathologically similar to those in human teeth, so bovine enamel is considered to be a suitable surrogate for human enamel in caries research.

Changes of PKCβII Protein Expression and Membrane Translocation in Pulmonary Artery of Rats with Hypoxic Pulmonary Hypertension

GUO Wan-jin, SHI Yi-wei, ZHANG Ai-zhen, DU Yong-cheng

2014, 34(3): 181-186. DOI: 10.3969/j.issn.1674-5817.2014.03.003

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Objective To establish rats model of chronic pulmonary hypertension (PH) and preliminarily explore the effect of PKCâII in the development of hypoxia-induced pulmonary hypertension. Methods Male SD rats were exposed to normobaric hypoxia to induce PH model. The rats were randomly divided into six groups, and exposed to normoxia control and to normoxia hypoxia for 1 d, 3 d, 7 d, 14 d and 21 d. Right ventricle systolic pressure (RVSP) and right ventricle hypertrophy index(RVHI) were measured. Pathological changes of the pulmonary arteries were observed by HE staining. The changes of PKCβII protein expression and membrane translocation in pulmonary arteries were also detected by Western blotting. Results RVSP increased significantly after hypoxic exposure for 1 d, 3 d, 7 d, 14 d and 21 d when compared with the normoxia control(P<0.05); There were significant rise in RVHI after hypoxic exposure for 3 d, 7 d, 14 d and 21 d (P<0.05). The pathological changes of the pulmonary arteries were significant at 3 d, 7 d, 21 d. PKCâII protein expression decreased significantly at 3 d, 7 d, 14 d, 21 d after chronic hypoxia than those of normal control groups respectively(P<0.05). Significant decrease of PKCβII membrane translocation level in pulmonary arteries of rats could be detected after hypoxic exposure 3 d(P<0.05). Conclusion The decrease in protein expression and membrane translocation of PKCβII may be involved in the development of the chronic hypoxia-induced pulmonary hypertension.

Establishment and Evaluation of Acute Kidney Injury Mice Model by Ischemia Reperfusion and Cisplatin Induction

JIANG Yan, WANG Wei, LI Ze-zheng, CHENG Jin, WANG Wei-wei, ZHANG Jin-yuan

2014, 34(3): 187-192. DOI: 10.3969/j.issn.1674-5817.2014.03.004

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Objective To evaluate renal tissue damage in acute renal injury model mice established by ischemia reperfusion (I/R) with different renal ischemia times and intraperitoneal injection of cisplatin with different concentrations. Methods The male C57BL/6 mice were randomly divided into the I/R group and cisplatin induction group. All the mice were sacrificed ,whose blood were sampled for detection of serum creatinine(Scr)and urea nitrogen(Bun), the kidneys samples were dyed with HE for observation of the pathological changes and evaluation of acute tubular necrosis score(ATN score). In situ renal tubular epithelial cells apotosis were detected by TUNEL. Results The levels of BUN and Scr and ATN score in I/R group and cisplatin induction group were much higher than those of in the sham group and control group (P<0.05). As to the situation where cisplatin concentration heightened and ischemia times prolonged, the apoptosis cells gradually increased, and the pathological damage in renal tissues was more severe. Conclusion Renal tubular necrosis occurred in some of the ischemia-reperfusion 45 min group. The ideal time of acute ischemia kidney injury is 30~40 min. 10 mg/kg intraperitoneal injection twice is a suitable dose to AKI model induced by cisplatin.The degree of renal tissue injury of the mice was more severely in ischemia-reperfusion group than cisplatin induced group.

Expression of Inflammatory Factors in Atherosclerosis Model of Bama Mini-pig

CHEN Fang-ming, PAN Yong-ming, CHEN Liang, SHI Yuan, ZHU Ke-yan, YU Xu-ping, CHEN Min-li

2014, 34(3): 193-198. DOI: 10.3969/j.issn.1674-5817.2014.03.005

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Objective To observe the expression of MCP-1, NF-κB p65, ICAM-1and VCAM-1 protein in the abdominal aorta and coronary of the atherosclerosis (AS) model of Bama mini-pig induced by high fat diet. Method Bama mini-pigs which were 4-5 months of age are divided into normal control group which are fed basal diet for 24 weeks normal and model group which are fed high fat diet for 24 weeks, each group included 5 ones. Then the abdominal aorta and coronary of these mini-pigs were taken to conduct oil red “O” fat stain and routine HE staining. The MCP-1, NF-κB p65, ICAM-1 and VCAM-1 protein expression were observed by Immunohistochemistry staining. Result The result of oil red “O” staining shows that arty streak deposition was visibly on the abdominal aortic in model group and HE staining showed that in model group, abdominal aorta obviously thickening which is type IV of AS lesions and the formation of atherosclerotic plaque in coronary artery. The result of Immunohistochemistry staining was that, to contrast with normal group, the MCP-1, NF-κB p65, ICAM-1and VCAM-1 protein expression in abdominal aorta of model group are significantly higher(P<0.05); the MCP-1, NF-κB p65, ICAM-1and VCAM-1 protein expression in coronary of model group are significantly higher(P<0.05) too. Conclusion Abdominal aorta and coronary artery lesions of the AS model of Bama miniature pig consistent with clinical pathological characteristics of human AS. The MCP-1, NF-κB p65, ICAM-1and VCAM-1 protein expression are rise in different degree, which consistent with the clinical AS inflammatory reaction and mechanism.

Comparison on Histology of Cardiovascular System in 6 Species of Laboratory Animals

WANG Yuan-yuan, XU Wen-mang, LI Xia, LI Tao, DAI Fang, YANG Ju-lun, SONG Shu-ling, LIU Qiang-gao

2014, 34(3): 199-204. DOI: 10.3969/j.issn.1674-5817.2014.03.006

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Objective To observe the differences on histology of the cardiovascular system in 6 commonly used laboratory animals. Methods Six species of laboratory animals, including 30 rhesus monkeys, 20 KM mice, 20 SD rats, 18 Japanese white rabbits,16 Beagle dogs, 20 tree shrews were selected. Except the mice were killed by cervical dislocation, other animals were sacrificed by intravenous anesthesia, and the heart and some blood vessels were removed, and then were fixed by 10% formalin and sealed in glass containers after pathological anatomy. The heart and blood vessels were processed by gross examination, routine pathological HE staining, necessary special staining and immunohistochemical staining, then the similarities and differences of cardiac and vascular structure and cell structure were observed under the microscope. Results (1) Six species of animals have no significant differences in the structure of the heart, the difference only exist in myocardial cell morphology. (2)Cardiac interstitial collagen deposition situation is different in the six species of animals. (3) The distribution of nutrient vessels among myocardial fibers are different in 6 species of animals. (4) Six species of animals have no significant difference in arterial vascular structure, only the thickness of tunica media and the content of collagen and elastic fibers are different. Conclusion The comparison on histopathology of cardiovascular system in six species of animals provides a reference for teaching and research works,and also provides appropriate basis and control for the development of“experimental animal pathology testing standards”.

Effects of Sleep Deprivation on Rat Liver Iron Metabolism

GUAN Yu, YAN Xiao-li, WANG Guo-hua, ZHANG Xiao-feng

2014, 34(3): 205-209. DOI: 10.3969/j.issn.1674-5817.2014.03.007

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Objective To explore the influence of sleep deprivation on rat liver iron metabolism and related molecular mechanism. Methods Twenty Sprague-Dawley male rats (weight about 180 g) were randomly divided into normal control group (CG) and sleep deprivation group (SDG). The SD model was induced in rats by improved multi-platform sleep deprivation method. After 5 days’ sleep deprivation, the colorimetry was used to detect the iron ion concentration in rat’s serum and liver tissue samples; the expression of HAMP, IL-6 and IL-1 mRNA in rat’s liver was observed by real-time fluorescent quantitative PCR. The expression of hepatic ferritin and STAT3 protein was detected by Western Blot. Results Compared with CG group, serum iron level in SDG rats reduced (P<0.05), while liver iron concentration was higher (P<0.01); the expression of HAMP, IL - 6 and IL - 1 mRNA in SDG liver tissue increased by 1.1, 3.2 and 1.7 times respectively (P<0.01); the expression of ferritin and phosphorylated STAT3 protein in SDG liver tissue significantly higher than control group (P<0.01). Conclusion Sleep deprivation disturb iron metabolism in rats through IL - 6 - hepcidin axis.

Effects of Anshen Dingzhi Ling on Spontaneous Hyperactivity,Anxiety and Space Learning and Memory in Attention Deficit Hyperactivity Disorder Rats

MIAO Jing, LIU Cheng-quan, HAN Xin-min

2014, 34(3): 210-213. DOI: 10.3969/j.issn.1674-5817.2014.03.008

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Objective To explore the effects on brain monoamine neurotransmitter levels and potential action of mechanism of Anshen Dingzhi Ling(ADL) in the treatment of attention deficit hyperactivity disorder (ADHD) rats. Methods Forty SHR rats were randomly divided into five groups: ADHD model group, Atomoxetine groups, ADL-low-dose groups, ADL-medium-dose groups, and ADL-high-dose groups, at the same time eight WKY rats were set as the normal control groups. All the rats were gavaged for four weeks in a row. The open field, elevated plus maze (EPM), Morris water maze were employed to observe the changes in the animal behavior before intragastric administration and 3 weeks after the administration. Results In open field test, ADL low-dose group (P<0.05) and Atomoxetine group (P<0.01) reduce the total distance of rats, and both of groups can increase the number of cross platform (P<0.01) in Morris water maze test .At the same time ADL can reduce the frequency and Residence time that rats into the open arms (P<0.01) in elevated plus maze. Conclusion ADL decreased the spontaneous hyperactivity, relieve anxiety behavior and improve the space learning and memory.

Effects of Corn Peptide and Aerobic Exercise on Levels of Glucagon、MCP-1、PP and PYY in Plasma of Obese Rats

LI Xiang, SHI Reng-fei, LOU Shu-jie

2014, 34(3): 214-218. DOI: 10.3969/j.issn.1674-5817.2014.03.009

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Objective To investigate the effects of corn peptide and aerobic exercise on the Glucagon, MCP-1(monocyte chemoattractant protein -1), PP(Pancreatic polypeptide) and PYY (Peptide YY) plasma levels for analyzing the hormonal related regulation of the body weight lose in obese rats. Methods Fifteen male SD rats were randomly selected from 150 as normal diet group (C). The rats of C were given a normal diet , while the remaining 135 were given a high fat diet. 8 weeks later the 40 male obese rats were randomly divided into obesity control group (OC) , casein supplement group (Cas), corn peptide supplement group (CP) , obese rats exercise group (OE) and corn peptide supplement combined obese rats exercise group (OEC). OE and OEC group rats took aerobic exercise during 4 weeks intervention. ELISA method was used to test the Glucagon, MCP-1, PP and PYY levels in the rats plasma. Results Four weeks intervention later, the body weight of OC was significantly higher than C, and the body weight of OE and OEC group were significantly lower than OC. There was no significant difference between OC and C in Glucagon level, but the Glucagon level of CP was significantly lower than OC. The MCP-1 level in OC group was significantly higher than C. and compared with OC, the MCP-1 levels of CP, OE and OEC decreased significantly . The PYY levels of OC were significantly lower than C, the PYY level of OEC was significantly higher than OC and CP. There was no significant difference among all groups for PP plasm level. Conclusion Corn peptides can reduce the plasma levels of Glucagon and MCP-1 in obese rats. The aerobics combined with corn peptide supplyment can reduce the plasma level of MCP-1 and increase the plasma level of PYY in rats.

Preparation of Magnetic Iron Oxide Nanoparticles and Their Distribution in Rat

ZHAO Wen, ZHANG A-na, ZHAO Qiang

2014, 34(3): 219-222. DOI: 10.3969/j.issn.1674-5817.2014.03.010

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Objective To research the preparation of magnetic iron oxide nanoparticles and their distribution in the SD rat. Methods A organic phase process is used for making high-quality monodisperse hydrophobic Fe₃O₄nanocrystals through the high temperature decomposition of Fe(acac)₃ and a long-chain alcohol in autoclave reactor. Then water-soluble iron oxide nanoparticles (WION) were prepared by mixing with a bipolar surfactant,2,3 dimercaptosuccinic acid (DMSA). Results The nanoparticles with mean sizes 10 nm are Fe₃O₄nanocrystal. 24 h after tail vein injection of WION 0.5 ml(1mg/ml solution), the WION mainly concentrated in the spleen and liver, kidneys and lungs followed. Conclusion Successfully prepared Fe₃O₄ nanoparticles through pyrolysis, and it has good surface morphology. Further modificated its surface by DMSA, can obtained a good dispersion of the WION.

Study on Gut Hormones in GK Rats after Different Length Duodenum-Preserving Duodenal-Jejunal Bypass Surgery

CHEN Li-Xin, ZHAO Xian-zhe, QIAO Wei-wei, GU Jian-Zhong, WANG Long-Xia

2014, 34(3): 223-227. DOI: 10.3969/j.issn.1674-5817.2014.03.011

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Objective To establish the different length duodenum-preserving duodenal-jejunal bypass model in GK rats and explore the changes in gut hormones after surgery. Methods Male Goto-kakizaki diabetic rats(GK, n=24) were randomly divided into four groups: group I (sham group), group II(regular DJB surgery), group III (8mm duodenum-preserving DJB surgery), group IV(18mm duodenum-preserving DJB surgery). Carotid artery intubation was performed ninth week after surgery. OGTT was performed one week after carotid artery intubation . The change of the level of the serum index (blood glucose, insulin, GLP-1, GIP) during OGTT was measured. Results All GK rats after surgery were with a long-term survival. The OGTT, first-phase insulin and GLP-1 secretion in group III were improved significantly compared with group I (P<0.05) and the initial secretion of GIP in group III was significantly fewer than group I (P<0.05). The OGTT, first-phase insulin and GLP-1 secretion , the initial secretion of GIP in group IV had no significant difference compared with group I (P>0.05). The first-phase insulin and GLP-1 secretion in group II were significantly higher than group IV (P<0.05) and the GIP secretion at 30 min was significantly higher than group IV (P<0.05). Conclusions On the basis of experimental results, the mechanisms of DJB surgery improving blood glucose level in GK rat might be closely related to the restoration of balance between anti-incretin factors and incretin factors. On the other hand, DJB surgery can improve insulin-resistance in GK rat through reducing the secretion of GIP.

Hypolipidemic Effect of Combination of Berberine and Metformin on Experimental Hyperlipidemia Models

MA Yi, CAO Jun-ping, XU Li, LI Xia, LI Dong-mei, SUN Hui-ping

2014, 34(3): 228-231. DOI: 10.3969/j.issn.1674-5817.2014.03.012

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Objective To investigate the hypolipidemic effect of combination of berberine and metformin (B-M, 2∶1) on rats and mice with hyperlipidemia. Methods Rats model of hyperlipidemia was established by administration of high-fat diet for 28 days. The model rats were randomly divided into 5 groups, namely B-M high, medium and low dose groups (280 mg/kg, 140 mg/kg and 70 mg/kg, respectively), Metformin group (140 mg/kg), model group, and the normal rats as the contorl group (n=10). After 21 days of oral administration, the sera of all rats were collected to determine the levels of TG, TC, HDL-C and LDL-C, respectively. Mice model of hyperlipidemia was established by a caudal vein injection of Triton WR-1339 at a dose of 400 mg/kg. Then model mice were randomly divided into 5 groups, namely B-M high, medium and low dose groups (400 mg/kg, 200 mg/kg and 100 mg/kg, respectively), Metformin group (200 mg/kg), model group, and the normal mice as the control group (n=10). All mice were orally administered pre-injection for 3 days. The sera of all groups were collected to detect the levels of TG, TC, HDL-C and LDL-C 24 h post-injection. Results Administration of B-M combination significantly reduced the TG, TC and LDL-C levels in serum of rats with hyperlipidemia, compared to the model group (P<0.05 or P<0.01). At the same time, B-M reduced the levels of blood lipids in mice with Triton-induced hyperlipidemia. The levels of TG, TC, HDL-C and LDL-C in high dose group, TG, TC and LDL-C in medium dose group and LDL-C in low dose group were significantly lowered than the model group (P<0.05 or P<0.01). In addition, a dose-dependent manner was showed in rats and mice with hyperlipidemia, respectively. Conclusion B-M combination may have a good hypolipidemic effect.

Effects of Astragalus Polysaccharides on B16-BL6 Melanoma Lung Metastasis in Mice

ZHAO Yong, ZHANG Lu, CAI Li-ping, XU Chen

2014, 34(3): 232-234. DOI: 10.3969/j.issn.1674-5817.2014.03.013

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Objective To explore the effect and mechanism of different doses of astragalus polysaccharides on pulmonary metastasis of B16-BL6 melanoma in mice. Methods The B16-BL6 artificial pulmonary metastasis model was used to study the influence of different doses of astragalus polysaccharides on pulmonary metastasis of melanoma; exploring its possible mechanism by measuring the immune organ index variation after administration in mice. Results High dose of astragalus polysaccharide can significantly inhibit B16-the BL6 melanoma lung metastasis and increase the index of spleen in mice. Conclusion High dose of astragalus polysaccharides maybe inhibited the pulmonary metastasis of melanoma cells by enhancing immunity.

The Research Progress on Simian Cytomegalovirus

WANG Sha-sha, HE Zheng-ming

2014, 34(3): 242-248. DOI: 10.3969/j.issn.1674-5817.2014.03.016

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Simian cytomegalovirus(SCMV) were the prototypic members of the beta herpesvirus subgroup, a group of DNA virus with segment rounded. Like all herpesvirus, they shared the similar structure characteristics, and could initiate persistent latent infection. Recently, SCMV were wildly concerned as the best suited model for human cytomegalovirus (HCMV) research, and the study of infection route, pathogenic mechanism and diagnosis and treatment for SCMV could lay a foundation for those of HCMV. This article reviewed the discovery , classification, biological characteristics, infection characteristics and immunological response, detections of SCMV, vaccines reserch and development prospects so as to provide some useful insights for researchers.

Advances in Research of SPF Rabbits

GUO Zhong-kun, WANG Ke-zhou, LV Su-jun

2014, 34(3): 249-254. DOI: 10.3969/j.issn.1674-5817.2014.03.017

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The study of SPF rabbits has lasted more than 50 years from the late 1950s. In this paper, the breeding, biological characteristics, feeding and management, and applied research of SPF rabbits were made a detailed exposition to have a better understanding about the development status of SPF rabbits at home and abroad, and to provide more abundant information for future study.

Experimental Operations Induced Stress in Rodents

WEI Shan-shan, YANG he-jin, QU Huihua, ZHAO Yan, WANG Qing-guo

2014, 34(3): 255-258. DOI: 10.3969/j.issn.1674-5817.2014.03.018

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The routine experimental operations ,such as handling, orogastric gavage, anaesthesia, blood collection, restrict, transport, etc . can induce stress in animals. The operations impact on some parameters correlated with stress and metabolic, which have a probable influence on experiment results. However, the impacts are usually ignored by researchers due to the pervasiveness of the operations. This article described the impacts respectively.

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