Table of Content

25 December 2012, Volume 32 Issue 6

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Comparative Study on Vascular Dementia Model Between SHR/SP and Wistar Rat

ZENG Gui-gang, ZHANG Shen, GU Jian-zong, CHEN Guo-qiang

2012, 32(6):  461-466.  DOI: 10.3969/j.issn.1674-5817.2012.06.001

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Objective To evaluate the effect of permanent hypertension combined with hypoperfusion on the Stroke Prone spontaneously hypertensive rat (SHR/SP) vascular dementia model. Method Male SHR/SP and Wistar rats were randomly divided into SHR/SP sham operation group,SHR/SP model group, Wistar sham operation group and Wistar model group respectively. All rats enrolled in the operation group were undergone a permanent ligation of bilateral carotid artery(2-VO).After the operation, 10 survival rats was available in each group. After 30 days, Morris maze tests were conducted for valuating the learning- memory abilities. Morphological changes of neuron cells and micro-blood vessels was observed. Result In place navigation training, the latency of Wistar sham operation group were less than that of rats in SHR/SP sham operation group, the latency of Wistar model group were less than that of rats in SHR/SP model group, the latency of SHR/SP model group were less than that of rats in Wistar model group (P<0.05). In spatial probe ability test, the trails of rats in SHR/SP and Wistar sham operation groups were concentrated upon platform quadrant, while the those of SHR/SP and Wistar model groups were disorgnized. Neurons loss and micro-blood vessels stenosis of the rats in SHR/SP and Wistar model group groups were severer than that of the rats in SHR/SP and Wistar sham operation groups in the brain region. Conclusion The cognitive function of SHR/SP vascular dementia model was serious than Wistar rat vascular dementia model, both chronic permanent hypoperfusion and hypertension may lead to the loss of neurons and myelin which may result in memory dysfunction. Hypoperfusion is even more harmful than hypertension.



Assessment of Autonomic Nervous Function in Sub-health Rats Induced by Different Factors Based on Heart Rate Variability Analysis

PAN Yong-ming, LIU Rui-min, Wu Wei, WANG Hui, JIA Lin-chao, XU Xiao-ping, ZHU Ke-yan, CHEN Min-li

2012, 32(6):  467-472.  DOI: 10.3969/j.issn.1674-5817.2012.06.002

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Objective To assess autonomic nervous function in sub-health rats induced by different factors based on Heart Rate Variability (HRV) analysis, and determine the relationship between HRV indexes and sub-health state. Methods Fourty-two male SPF Wistar rats were randomly divided into seven groups: multi-factor group (MF), multifactor treatment group (MFT), warm swimming group (WS), dietary restriction group (DR), sleep deficiency group (SD), purely constraint group (PC) and control group (C), six animals in each group. Using warm swimming, dietary restrictions, sleep deprivation and constraint to establish sub-health rat model. Rats were daily orally administered 300 mg/kg anti-aging tablet in MFT group. The ECG was monitored at the end of the modeling and recovery of 3 days, and HRV analysis has been used to investigate the autonomic nervous function in sub-health rats induced by different factors. Results Compared with control group, standard deviation of the mean R-R interval (SDNN), the root-mean-square of successive differences (RMSSD), short term variability (STV) and long term variability (LTV) parameters of their HRV time domain analysis in sub-health rats were significantly higher than those of control group, and very low frequency power (VLF), low frequency power (LF), total power (TP), LF/HF ratio, normalized low frequency (LFnu) parameters of their HRV frequency domain analysis were increased, while normalized high frequency (HFnu) was decreased. Anti-aging tablets treatment could significantly reduce the LF, LF/HF ratio and LFnu (P<0.05) and increase HFnu (P<0.05). Univariate logistic regression analysis showed that the closely relationship between RRI, SDNN, LTV, LF, TP, LF/HF ratio, LFnu, HFnu and sub-health state (P<0.05), and stepwise logistic regression analysis showed that the low HFnu (OR, value=0.566, P=0.023) maybe independent risk factors for inducing sub-health. Conclusion The autonomic nervous function in sub-health rats induced by different factors was apparent disorder, and inhibition of vagus nerve activity maybe an independent risk factor for sub-health. So HRV analysis could assess the sub-health state.

Establishing a New bone- tendon Junction Injure Model in Rat Facilitating MRI Detection

LIANG Pei-hong, LI Ai-guo, HE Ming-chong, ZHONG Ming-zhi, CHEN Shong, LIANG Zhi-ping, YANG Xiao-hong, WANG Wen, CHEN Hong-hui

2012, 32(6):  489-492.  DOI: 10.3969/j.issn.1674-5817.2012.06.007

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Objective To establish a new bone-tendon junction injury animal models for better observation with MRI. Methods The patella of 12 mature New Zealand white rabbits were partially resected and then the tendon were sutured to bone.General examination and MRI were done after 2, 4, 8 and 12 weeks. Results The general examination and MRI results showed the healing process of bone-tendon junction dynamically in 2, 4, 8, 12 weeks after operation. The process was featured as cancellous bone regeneration and bone- tendon healing contact fibrocartilage zone regeneration, which was restored gradually as time pasted. Conclusions The modified animal models do not affect bone-tendon junction healing and is better for MRI observation correctly.



The Expression of α-SMA, MMP13 and Model Evaluation in uPA Knock-out (uPA^-/-) Mice with Liver Fibrosis

XIA Min-jie, CHENG Qian-qian, WANG Yu-zhu, TIAN Fang, WEN Cheng-li, WANG Xiao-dong, LI Wei-hua, DING Xun-cheng

2012, 32(6):  493-498.  DOI: 10.3969/j.issn.1674-5817.2012.06.008

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Objective To evaluate the feasibility of uPA knock-out (uPA^-/-) mice liver fibrosis model by analysing the expression of α-Smooth Muscle Actin (α-SMA) and Matrix Metalloproteinase-13 (MMP13). Methods Adult male C57BL/6J WT mice and uPA knock-out (uPA^-/-) mice were divided into four groups with 10 mice in each group: control groups (Con-WT, Con- uPA^-/-) and liver fibrosis model groups (Mod-WT, Mod-uPA^-/-). Mice were intraperitoneally injected with 0.15ml 10% CCl₄ (or olive oil as control) twice per week for 6 weeks to induce liver fibrosis. The mRNA expression of á-SMA and MMP13 was detected by real-time polymerase chain reaction (PCR) and the protein expression of α-SMA and MMP13 was analyzed by western blot and immunohistochemistry. Result The real-time PCR showed that the mRNA expression of α-SMA, MMP13 in uPA^-/- mice wes significantly higher than that in WT mice. The western blot results indicated that there were no expression of α-SMA protein and MMP13 protein of the mice liver in the control group, a few expression of α-SMA protein in Mod-WT group, and a significant increase of α-SMA protein expression in the Mod-uPA^-/- group. The immunohistochemistry results demonstrated that the protein expression of α-SMA and MMP13 of Mod-uPA^-/- group was higher than Mod-WT group. Conclusion The mRNA and protein expression of α-SMA were significantly higher in the uPA^-/- mice. The knocking-out of the uPA gene promoted the hepatic stellate cells transforming into myofibroblasts, thus speeding up the deposition of extracellular matrixin in the liver. Also, the protein expression of MMP13 in liver fibrosis model mice was significantly increased. uPA^-/- mice are susceptible animal strain to establish liver fibrosis model.



Effect of Bone Marrow Mesenchymal Stem on Deep Partial Thickness Derma Burn Injury in Hamsters

MA Mei, ZHANG Yan, WEI Qin, JIANG Tao, DUAN Min-jun, ZHANG Chun, WANG Xun-mei, HOU Yue-mei

2012, 32(6):  499-502.  DOI: 10.3969/j.issn.1674-5817.2012.06.009

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Objective To investigate the effect of bone marrow mesencymals stem cells transplantation on the deep partial thickness derma burn injury in hamsters and explore the possible mechanism. Methods Fourty hamsters were used to create deep partial thickness derma burn injury model and randomly divided into two groups: transplantation group (group A) and control group (group B). The wound of animals in group A were treated with local transplantation of the fifth generation cells of MSCs cultured in vitro with concentration of 1×10⁷ /ml plus surface daub of supernatant. The group B were treated with the same amount of saline. The wound healing effect was observed by HE staining, chymase mRNA level was analyzed on day 1, 3, 7, 14 after transplantation. Results The HE staining results showed the quality of healing was better in transplantation group than in control group. PCR results showed that chymase gene expression was found in both groups with the highest expression on day 3.Conclusion BMMSCs transplantations can promote wound healing, and dermal mastocyte chymotrysin would be involved in wound healing process.



Animal Experimental Studies of Power Frequency Electromagnetic Field Exposure on Reproductive Function of Rats

YU Lin, SHI Yan, XU Yan, WANG Wei-feng, MIAO Mao-hua, SHI Hui-juan, SUN Zhao-gui, YUAN Wei

2012, 32(6):  503-507.  DOI: 10.3969/j.issn.1674-5817.2012.06.010

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Objective An instrument of power frequency electromagnetic fields, applicable for an animal model with rats as experimental subjects, will be designed to evaluate risk of the damage on reproductive functions under low frequency electromagnetic field exposure. Method The rats were exposed to the magnetic field at certain intensity for some time. Some parameters of reproductiue function were measured including plasma levels of steroid hormones, estrous cycle, and the numbers and development status of the offspring in females, and weight of the testis and epididymis, sperm density, the proportion of motile sperms, and sperm plasma membrane integrity. Results The study found that one month exposure of 1mT electromagnetic field, 8hours daily, initiated at the age of 4 weeks didn’t induced significant change in the secretion of hormones and fertility in females, only increased estrus frequency, in the power frequency electromagnetic field at 1mT, and with daily exposure time of 8 hours, for a month since the beginning at age of 4 weeks, the grown adult female rats have not shown a significant change in the secretion of hormones and fertility to bear future generations, and only estrus frequency increased. two month exposure of 2mT electromagnetic field, 12hours daily, initiated at the age of 8 weeks induced the decline of sperm motility of males.



Comparative Research on Two Skin Infection Models in SKH-1 Hairless Mice Induced by MRSA

CHEN Xue-yu, PENG Xiu-hua, ZHU Ping-mei, XU Chun-hua, ZHOU Wen-jiang, ZHOU Guang-xing

2012, 32(6):  508-511.  DOI: 10.3969/j.issn.1674-5817.2012.06.011

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Objective To explore methods to establish different models infected with methicillin resistant Staphylococcus aureus (MRSA). Methods Forty-eight specific pathogen free SKH-1 hairless mice were randomly divided into 4 groups. Mice from the PBS control groups were treated with sterilized PBS, by subcutaneous injection and direct daub on the puncture of skin injury by needle respectively, mice from model groups were treated with MRSA (ST-239) by subcutaneous injection or direct daub on the puncture of skin injury by needl. Models were evaluated by appearance and volume of abscesses or ulcers, CFU extracted from skin and morphology. Results There were significant differences in appearance, volume and CFU extracted from skin between the two models. Evident pathological alterations were observed. Skin of mice from subcutaneous injection group developed abscesses with limited inflammation, with no abscesses and diffuse inflammation induced in needle injury group. Conclusions Mice treated with MRSA by subcutaneous injection or diret daub on the puncture of skin injury by needl can develop abscesses model and wound-infected model respectively.



Establishment of Blood Glucose Fluctuation Model on Diabetic Rats and Its Damage to Pancreas

ZHU Yan-tao, JIN Rong-jia, ZHONG Jie-min, LI Shou-ye, XU Zhi-wei, SHOU Qi-yang, CAI Yue-qin, WANG Hui

2012, 32(6):  512-516.  DOI: 10.3969/j.issn.1674-5817.2012.06.012

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Objective To establish the animal model of diabetic rats with fluctuant high blood glucose and observe the effects of fluctuant high blood glucose on pancreas in diabetic rats. Methods The diabetic rats model was developed by injection with streptozotocin (STZ) 30 mg/kg, regularly staggered insulin 1U and glucose 0.375 mg/kg administration were used to set up blood glucose fluctuation model. General symptoms were observed and the data of FFA、INS、HbAlc in blood as well as the Bax and Bcl-2 in pancreas were measured at the thirteen week. Results (1) The typical diabetic symptoms in blood fluctuation group and diabetic group were more obvious than those in normal group. (2) The blood glucose levels in diabetic group and blood glucose fluctuation were significant higher than that in normal group. (3) The FFA、HbAlc、INS in diabetic group and blood glucose fluctuation group were higher than those in normal group (P<0.01); The FFA、HbAlc、INS in blood glucose fluctuation group was higher than those in diabetic group (P<0.01). (4) Compared with normal group, the Bax in pancreas was obviously increased (P<0.01) and the Bcl-2 was dramatically reduced in blood fluctuation group and diabetic group (P<0.01); The Bax was higher (P<0.01) and the Bcl-2 was lower (P<0.01) in blood fluctuation group than those in diabetic group. Conclusion The blood fluctuation diabetic rats model was successfully established by the method of subcutaneous insulin and intraperitoneal glucose injection in diabetic rats twice daily, and fluctuant high blood glucose aggravates the injury of the pancreas in diabetes rats.



The Effect of Carbamazepine on Cardiac Function of Zebrafish Embryo

WANG Xue, WANG Xi-min, HAN Li-wen, LIU Ke-chun, HE Qiu-xia, PENG Wei-bing

2012, 32(6):  517-520.  DOI: 10.3969/j.issn.1674-5817.2012.06.013

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Objective To explore the feasibility of studying drug toxic and side effect with zebrafish embryos as model. Methods Fourty-eight hours post-fertilization zebrafish embryos were treated with various concentrations of carbamazepine for 24 h. Morphological changes of embryos hearts were observed and heart rate was counted at 8, 16, 24 h after treatment. Then embryos were transferred into embryo medium and the heart rate was observed at 4, 8, 24 h after treatment interruption. Results The heart rate decreased significantly after treatment with carbamazepine for 24 h and showed a good dose-effect relationship with the drug concentration. The 24 h EC₅₀ of decrease of heart rate was about 626.07 µmol/L. Heart rate recovered quickly 8h after treatment interruption.Conclusion Carbamazepine lowered the heart rate of zebrafish embryo in a dose-dependent manner and the repression on cardiac function was reversible.



Comparative Study on Several Biological Characteristics of Clean Rats in Three Rearing Environments

HU Ying, YANG Fei, WU Jian-ping

2012, 32(6):  521-526.  DOI: 10.3969/j.issn.1674-5817.2012.06.014

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Objective To study the effects of different rearing environment, barrier facility+IVC, barrier facility+conventional cage and conventional environment facility+IVC on the stress level, immune function and hematology and blood biochemistry in clean rats. Methods Three groups of weanling clean rats with 3 in one cage were housed respectively in three different environments for 45 days before tests of neuroendocrine hormone, cytokines, hematology and blood biochemistry were conducted, and the results were compared by groups to discover the different effects of three rearing environment. Results Compared with the rats in barrier facility+conventional cage, the rats in barrier facility+IVC had lower levels of CORT, IL-2, IL-1α, TNF-α, WBC, IL-1β, Fractalkine, CREA, and higher levels of IL-1β, Fractalkine, BUN, Ca, Ca/P ratio, meanwhile, the rats in conventional environment facility+IVC had lower levels of CORT, MCP-1, TNF-α, GLU, CREA, and higher levels of IL-1β, Fractalkine, RBC, HGB, HCT, TP, ALB, UA, Ca, Ca/P ratio, K, CO₂. Moreover, rats housed in IVC in different environment facility also showed significant difference in IL-1β, IL-10, MCP-1, RBC, HGB, HCT, BUN, IL-1α, ALB, ALT, UA and K. Conclusions Acclimatization of clean rats to different rearing environment can cause diversity of stress background, cytokine baseline and hematology and blood biochemistry. It suggests choosing a good rearing environment base on the rearing goals, periods and the relative interfering factors to be avoided.

Progress on Monitoring Methods of Oxidative Stress after Stress in Laboratory Animals

ZENG Xian-cheng, MA Jing, CHANG Yan, LIU Quan-hai

2012, 32(6):  549-552.  DOI: 10.3969/j.issn.1674-5817.2012.06.021

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Stress could induce oxidative stress in laboratory animals. Changes in energy metabolism in several aspects of the organism are one of the mechanisms through which the stress induces oxidative stress. Glucocorticoids take part in occurrence of oxidative stress after stress. Reactive oxygen species(ROS) levels, antioxidant capacity and contents of biomarkers of oxidative stress injury can be used to evaluate occurrence and damage of the oxidative stress. It was reviewed in this article that progress in the fields of how strees induce oxidative stress in laboratory animals, the mechanisms and the monitoring methods in recent years.