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Improvement of the resistance exercise apparatus of climbing ladder with load on rats

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):450-454. doi:10.3969/j.issn.1005-4847.2017.04.018

Objective To improve the resistance exercise apparatus of climbing ladder with load on rats, and to observe the technological, athletic training and biological response to the improved experimental apparatus.Methods Thirty-six healthy SPF male Sprague-Dawley were randomly divided into control and exercise groups.The climbing ladder and weight bearing device were separately improved to develop a new resistance exercise apparatus of climbing ladder with load on rats.The exercise group took an 8-week incremental exercise program using this improved device.To observe the operation effect of this improved exercise apparatus.To observe the dynamic changes of average maximum load on SD rats from the exercise group during 8 weeks.To compare the effect of climbing ladder exercise with load on body weights of SD rats between the two groups.Results The improved climbing ladder was marked by the prioritization technique of architecture stabilization, weight bearing and easy cleaning.The improved bearing device has the advantages of being sturdy and durable, and simplemethod to calculating load.Moreover, the new experimental apparatus can be operated by one person during the exercise program.The average maximum load was successfully increased to 756 g after an 8-week exercise using the improved apparatus.Compared with the control group, the body weights of SD rats from the exercise group after training were significantly reduced (P<0.05).Conclusions The improved resistance exercise apparatus of climbing ladder with load can provide a more endurable and repeatable process in the study of rat strength training and it is recommended to use this new device in future experimental research.

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Effects of glucose supplementation on the activity of myocardial AMP-activated protein kinase in rats induced by exercise

Haiyan YU ; Weiyu GU ; Zhiqiang WANG ; Zhengli YAO ; Jia ZHAO ; Rui ZHU ; Yunhong WANG

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):444-449. doi:10.3969/j.issn.1005-4847.2017.04.017

Objective To investigate the effect of glucose supplement on AMPK activation in myocardium of exercised rats by measuring the myocardial AMPK activation and glycogen content after acute exercise training.Methods Rats were subjected to an acute endurance exercise and glucose supplement in varying doses and time points before and after exercise.The dynamic changes of myocardial AMPK activities was measured with Western blotting, changes of myocardial glycogen content were measured with Anthrone method.Results AMPK activation in myocardium of exercised rat was increased significantly throughout the exercise, and remained at a higher level 1 hour after acute exercise.However the level of AMPK activity was not significantly increased in exercised rat with glucose supplement.Glycogen content was not significantly changed after exercise.Rats subjected to lower dose glucose supplement did not show significant changes in glycogen content neither.But glycogen content was significantly increased in rats at 24 hours after exercise, subjected to higher dose of glucose supplement.Conclusions 1) Acute exercise induces a significant increase in AMPK activation in myocardium of exercised rats.Glucose supplement significantly inhibites the activation of AMPK induced by acute exercise.(2) Higher dose glucose supplement significantly increases glycogen content in the rat myocardium 24 h after exercise.

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Application of implantable telemetry and whole-body plethysmography to observe the changes of circadian rhythm in conscious rats and its application in pharmacological safety evaluation of doxofylline

Shuizhen PAN ; Lizong ZHANG ; Yongming PAN ; Dejun WANG ; Yuanyuan LI ; Jiajun SHI ; Qiyang SHOU

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):438-443,449. doi:10.3969/j.issn.1005-4847.2017.04.016

Objective To study the application of implantable telemetry and whole-body plethysmography to observe the changes of circadian rhythm in conscious rats and evaluate the pharmacological safety of doxofylline, and to provide a basis for the future application of this technological system for drug safety evaluation.Methods Eight healthy SPF Sprague-Dawley rats were used in this study, 4 males and 4 females.The rats were implanted with telemetry transmitters by surgery to establish a telemetry system combined with plethysmography to observe the changes of 24 h physiological parameters and circadian rhythm in conscious rats at 14 d after operation, including heart rate (HR), blood pressure, the time interval from the Q wave to point A in the ECG of the aortic pressure wave (QA interval), respiration, activity, body temperature and pulmonary function parameters.The rats were divided into 3 groups: normal control group, doxofylline 40 mg/kg and 80 mg/kg groups, and the performance was validated by aerosolizing saline, doxofylline 40 mg/kg and 80 mg/kg inhalation, respectively, to observe the changes in physiological parameters after the drug administration.Results The physiological parameters of rats showed obvious changes in circadian rhythms at 14 d after operation.Compared with the normal control group, the doxofylline 40 mg/kg-treated group showed significantly increased changes of HR, tidal volume (TV), minute ventilation (MV), 50% expiratory flow (EF50), peak inspiratory flow (PIF) and peak expiratory flow (PEF) (P<0.01), significantly decreased respiratory frequency, QA interval and enhance pause (Penh) (P<0.05, P<0.01), but no significant differences in the blood pressure, activity and body temperature (P>0.05).Compared with the normal control group, the group treated with doxofylline 80 mg/kg had significantly increased HR, blood pressure, TV, MV, EF50, PIF and PEF (P<0.01), significantly decreased respiratory frequency, QA interval and Penh not (P<0.01), but not significantly changed activity and body temperature (P>0.05).Conclusions The application of implantable telemetry and whole-body plethysmography in this study does not obviously affect the circadian rhythm, and can sensitively monitor the relevant cardiovascular and respiratory parameters in conscious rats.It can be used in drug safety pharmacological research of cardiovascular and respiratory systems in conscious rats.

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Comparison of the behavior characteristics in SHR, WKY and SD rat models of attention deficit/hyperactivity disorder

Rongyi ZHOU ; Jiaojiao WANG ; Xinmin HAN

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):380-385. doi:10.3969/j.issn.1005-4847.2017.04.007

Objective To compare the differences in behavior characteristics among SHR, WKY and SD rat models of attention deficit/hyperactivity disorder (ADHD), and explore an ideal control model of SHR rats.Methods Using open field test to analyze the rat movement distance, speed, wearing numbers and the number of grooming to evaluate the spontaneous movement in SHR, WKY and SD rats.Using the Morris water maze to test the learning and memory ability among the three rat groups.Results The result of open field test showed that the SHR rats had significantly increased (P< 0.01) total amount of exercise, average speed and wearing numbers than WKY and SD rats.Compared with the WKY rats, SD rats had a significantly higher movement distance (P< 0.01), slightly higher movement speed and wearing number (P< 0.05).In the Morris water maze hidden platform period test, the SD rats had a significantly longer latency than the SHR rats (P< 0.05).SD rats showed longer latency distance on the first, third and fourth days of training, as compared with the SHR rats (P< 0.05 or P< 0.01).Compared with the WKY group, SD rats showed a shorter latency distance in each training time (P< 0.05 or P< 0.01).In the probe trial period, the SD rats showed shorter time and distance ratio to the target quadrant than SHR rats (P< 0.05), while significantly longer than the WKY rats (P< 0.05 or P< 0.01).Conclusions There are significant behavioral differences between SHR and WKY rats, showing certain disadvantages in comparison of the two types of rats.To add SD rats as a control group for SHR rats can improve the comparability of behavior characteristics of SHR rats, and to get more objective evaluation of the behavior characteristics of SHR rats.

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Establishment of a tree shrew model of Fusarium keratitis

Yan LI ; Jiejie DAI ; Xiaomei SUN ; Hai LIU ; Zhulin HU

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):420-424. doi:10.3969/j.issn.1005-4847.2017.04.013

Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.

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An improved method for preparation and evaluation of a porcine model of acute myocardial infarction by minimally invasive surgery and left circumflex artery ligation

Qin LAN ; Zhulin WANG ; Hao CAO ; Xiyan HAN ; Yun DONG ; Hongming ZHU ; Huimin FAN

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):414-419. doi:10.3969/j.issn.1005-4847.2017.04.012

Objective To develop a better method for preparation of porcine model of acute myocardial infarction by permanent occlusion of the left circumflex coronary artery and minimally invasive surgery, evaluate its validity and stability, and explore its application in experimental studies of ischemic heart diseases.Methods 25 healthy female 3-month-old Bama minipigs, body weight 25±3 kg, were used in this study.The porcine model of myocardial infarction was established by minimally invasive surgery and the left circumflex artery ligation at the site of OM1 posterior position under general anesthesia.Heart function was assessed by echocardiography at 15 min before surgery, 1 hour and 4 weeks after surgery.Pathological examination was performed at 4 weeks after the left circumflex artery occlusion.The mortality and cause of death were statistically analyzed.Results The 1-hour and 4-week postoperative cardiac function was considerably decreased, showing a decreased ejection fraction from 64.2±4.6% to 48.2±5.3% (1hour after MI) and 49.7±6.1% (4 weeks after MI) (P<0.01).Pathological examination revealed that the ventricular wall was thinner and the amount of collagens was increased in the infracted area.The ventricular fibrillation rate at 1-hour after myocardial infarction was 17.3% and the infarction area was 19.2%.Conclusions A pig model of acute myocardial infarction can be prepared by our modified left circumflex coronary artery ligation at the obtuse marginal artery (OM1) and minimally invasive surgery.This model exhibits advantages such as minimal surgical trauma, high stability of the model, and low mortality, therefore, provides an ideal and economic animal model for experimental studies on acute ischemic heart diseases.

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Establishment of a mouse model of aorta dissection induced by β-aminopropionitrile drinking combined with angiotensin II infusion

Yuting LIU ; Yanxiang GAO ; Shanshan WANG ; Wei REN ; Weiliang SUN ; Changan YU ; Jingang ZHENG

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):399-403. doi:10.3969/j.issn.1005-4847.2017.04.010

Objective To establish a mouse model of aorta dissection (AD) by β-aminopropionitrile (BAPN) in drinking water + subcutaneously pumped angiotensin II (Ang II) infusion.Methods Forty 3-week-old C57B1/6J male mice were randomly divided into two groups.All animals received 0.1 g/kg/d BAPN in drinking water for 4 weeks.Then the BAPN drinking + saline infusion group and BAPN drinking + Ang II infusion group received continuous saline or Ang II (1,000 ng/kg/min) infusion, respectively, via subcutaneous osmotic minipump for 72 hour.The mice were restricted in a noninvasive computerized tail-cuff system and their arterial systolic blood pressure and heart rate were monitored.Autopsy was performed if a mouse died during the experiment.At the end of the experiment, mice were sacrificed by injection with an overdose of sodium pentobarbital and the aortas were harvested.The formation of aortic false lumen was observed by pathology using hematoxylin-eosin staining.Results The overall incidence of AD in the BAPN drinking administration +Ang II infusion group was 95%, whereas the incidence of AD in the BAPN drinking administration +saline infusion group was only 5%.The mortality from dissecting aneurysm rupture was 24% in the BAPN drinking administration +Ang II infusion group during the experiment.Pathological examination of the aortic cross-sections clearly showed the formation of blood-filled false lumens induced by Ang II.Conclusions A mouse model with high incidence of aortic dissection is successfully established.

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Proliferation-promoting effect of umbilical cord mesenchymal stem cells on co-cultured bovine mammary gland epithelial cells

Yankun ZHAO ; Wei SHAO ; Chenglong LUO ; Xiong YU

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):391-398. doi:10.3969/j.issn.1005-4847.2017.04.009

Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells (BMECs) co-cultured with umbilical cord mesenchymal stem cells (UC-MSCs) in serum-free culture mediuum.Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact.In the direct contact culture groups, UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶1, 1∶1, 1∶2, 1∶3, 1∶4, 1∶5, and 1:10, respectively.In the indirect contact culture group, the supernatant of UC-MSCs was used as the conditioned medium to re-suspend BMECs.In the control groups, UC-MSCs and BMECs were cultured alone.The cell growth status in each group was observed at 0, 4, 8, 12, 24, 36, 48, 60, 72 h after culture, and cell proliferation was detected by cell counting kit-8 (CCK-8) assay.Results At 48 h, the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups (P<0.05).Meanwhile, the optical density in the direct contact group at a concentration ratio of 1∶2 reached the peak, which was extremely significantly higher compared with the control groups (P<0.01) and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group (P<0.05).Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs, and the co-culture by cell-to-cell contact has a better effect.The optimal concentration ratio of UC-MSCs to BMECs is 1∶2, and the optimal culture time is 48 h.

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Comparison of mouse models of Lewis lung carcinoma subcutaneously transplanted at different sites

Xueman MA ; Mingwei YU ; Ganlin ZHANG ; Jie YU ; Kexin CAO ; Xu SUN ; Guowang YANG ; Xiaomin WANG

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):386-390. doi:10.3969/j.issn.1005-4847.2017.04.008

Objective Subcutaneous transplantation Lewis lung carcinoma model is commonly used in experimental studies.Researchers often choose different transplantation sites to create the models while little attention was paid on the effect of different inoculation sites on the formation of transplanted tumors.The aim of this study was to compare the effect of tumor cell inoculation at different sites on tumor formation in mice.Methods Lewis lung adenocarcinoma (ll2-luc-m38) cells stably expressing luciferase protein were subcutaneously injected into C57 BL/6 mice at the right armpit, right groin, or footpad, respectively.An IVIS spectrum in vivo imaging system was used to observe the tumor and metastasis formation.The survival time and mortality were recorded.H-E stained pathology was performed to examine the histological changes of the lung tissues and tumor metastesis.Results The tumor formation time was earlier in the armpit and groin groups, both with a tumor formation rate of 100%, while the tumors occurred later, with a tumor formation rate of 33% in the footpad group.The pulmonary metastasis rate was 70% in the groin group, 50% in the ampit group, and 0% in the footpad group, at the 21st day after inoculation.The footpad group had a high mortality.The tumors in the groin group and armpit group can be surgically resected, with a postoperative survival rate of 100%.Conclusions In this mouse model of subcutaneously transplanted Lewis adenocarcinoma, the groin and ampit groups have advantages such as a high tumor formation rate, good tolerance of tumor resection, low surgical mortality rate, easy to monitor, simple operation and high reproducibility.The axillary group has an even higher metastasis rate.

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Expression and purification of asprosin and its effects on cardiac function in mice

Liwen ZHU ; Yanzhen TAN ; Wenping LUO ; Wei YI

Acta Laboratorium Animalis Scientia Sinica.2017;25(4):368-372,379. doi:10.3969/j.issn.1005-4847.2017.04.005

Objective To purify asprosin protein expressed in Escherichia coli expression system and to study its effect on cardiac function.Methods Coding sequence of asprosin was obtained from GenBank.Codon optimization was performed according to the codon preference of E.coli.After gene synthesized, recombinant plasmid was made.Asprosin was then induced and purified by Ni-affinity purification.The mouse model of impaired cardiac function was established by ligating and relaxing the left anterior descending coronary artery.30 mice were randomly divided into 3 groups: sham operation group (sham), cardiac dysfunction group (MI/R) and cardiac dysfunction plus injection of recombinant asposin protein group (MI/R+rAsp).The left ventricular function was detected by echocardiography to determine the improving effect of recombinant asprosin protein on cardiac function.Results After prokaryotic expression and purification, the purity of the target protein was higher than 95%, and the endotoxin content was less than <0.1 EU/μg protein, which was suitable for cell and animal studies.After the recombinant asprosin protein was given, the left ventricular function of the mice was improved significantly (P<0.05).Conclusions Asprosin acts as a myocardial protective molecule to improve cardiac function.

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