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Effect of LAT1 on early placenta formation in mice

Jing MA ; Yi TAN ; Dongmei TAN ; Junjie LU ; Hao LIANG ; Wenping LUO

Acta Laboratorium Animalis Scientia Sinica.2015;(3):256-260. doi:10.3969/j.issn.1005-4847.2015.03.007

Objective To explore the expression and role of LAT 1 in mouse uterus on early placenta formation on day 8 of pregnancy (D8).Methods One hundred and twenty 6-8-week old SPF female Kunming mice were used in this study.Immunohistochemistry was applied to determine the localization of LAT 1 protein in the mouse uterus on D8 of preg-nancy.The ectoplacental cones (EPCs) were dissected out from D8.5 uterus, and then cultured in vitro with different con-centrations of BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, specific antagonist of LAT1) and L-leucine ( substrate of LAT1) to determine the role of LAT1 during the EPC attachment and outgrowth .Results LAT1 protein was highly expressed in secondary decidual zone and also positively expressed in the mouse uterus on D 8.As a specific antago-nist of LAT1, BCH significantly suppressed the ectoplacental cone outgrowth , whereas L-leucine showed no significant effect on it.Conclusions LAT1 is expressed in the mouse uterus during early placenta formation and promotes ectoplacen -tal cone outgrowth , suggesting that LAT1 may promote the trophoblast invasion into maternal decidual tissue , and partici-pates in the early formation of placenta .

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Differential analysis of the bacterial community composition in mouse feces and intestinal contents

Yaqi WU ; Genshen ZHONG ; Minna WU

Acta Laboratorium Animalis Scientia Sinica.2015;(3):249-255. doi:10.3969/j.issn.1005-4847.2015.03.006

Objective To explore the objectivity and scientificity of fecal sampling , and to provide reference for investigating the relationship between intestinal microbes and diseases . Methods Terminal restriction fragment length polymorphism, degeneration gradient gel electrophoresis and real time fluorescent quantitative PCR techniques were applied to differentially analyze the bacterial community composition and abundance of intestinal contents and feces taken from dif -ferent sites of BALB/c mouse intestine .Results The predominant T-RFLP fragments ( T-RFs) in feces in the rectum and colon were 244 bp, 255 bp and 449 bp, however , those in feces of the small intestine including duodenum , jejunum and il-eum were 60 bp, 73 bp, 261 bp, 268 bp and 272 bp, and with a larger variation of the bacterial community composition in various parts of the small intestine .The bacterial abundance in the contents of duodenum and jejunum were 6.9 log ( cop-ies)/g and 8.3 log (copies)/g, fewer than in the other parts of the intestine , while the bacterial abundance in the feces was as high as 11.8 log (copies)/g, being about 2 times higher than that in the duodenum and jejunum (P<0.05), and similar to that in the ileum and colon content (P>0.05).Conclusions The inter-mouse variations of bacterial communi-ty composition in the large intestine contents are small .The bacterial composition and abundance are similar suggest that studies on the relationship between large intestine especially colorectal microbiota and diseases may be conducted via fecal sampling.

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Detection of growth phenotype and blood biochemical parameters of wild type-derived chromosome 1 substitution mouse strain

Chuan GAO ; Wei XU ; Fuyi XU ; Yaoqi ZHANG ; Ying ZHAO ; Liya ZHAO ; Yuxun ZHOU ; Kai LI ; Junhua XIAO

Acta Laboratorium Animalis Scientia Sinica.2015;(3):233-238. doi:10.3969/j.issn.1005-4847.2015.03.003

Objective To analyze the growth phenotype and blood biochemical parameters of chromosome 1 substi-tution mouse strain(CSS1), and investigate their potential of QTL mapping .Methods Body weight, body length, tail length, organ weight of the CCS1 mice were measured at different days to create a growth curve while blood biochemical in -dexes were measured at about the 80th day.Results The CCS1 mice were different from C57BL/6 mice in several inde-xes.Compared with the C57BL/6 mice during different developmental stages , six strains including B6-Chr1KM mice were significantly different in body weight .There were five strains including B6-Chr1CM mice significantly different with C57BL/6 mice in body length, and all of the CSS1 mice were significantly different from C57BL/6 mice in tail length.Part of CCS1 mice were significantly different from C57BL/6 mice in the weight of liver, spleen, kidney and brain.The ALT of female B6-Chr1CM mice was significantly higher than that in the C 57BL/6 mice.The ALP of female B6-Chr1HZ mice was signifi-cantly higher than that in the male C57BL/6 and B6-Chr1KM mice, and was significantly lower than that in the C57BL/6 mice.The TB of male B6-Chr1CM, B6-Chr1SMX and B6-Chr1HZ mice was significantly higher than that of the C 57BL/6 mice.The TG of male B6-Chr1SMX mice and male B6-Chr1TW mice was significantly higher than that in the C 57BL/6 mice. Conclusions The phenotype of Chr1 CSS mice is quite different from commonly used inbred strain C 57BL/6 mice.CCS1 mice show great potential in QTL mapping for their characteristic growth phenotype and blood biochemical indexes .

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Establishment of a mouse model of early lung adenocarcinoma

Shiman XIAO ; Yu ZHANG ; Jinwei QIANG

Acta Laboratorium Animalis Scientia Sinica.2015;(3):227-232. doi:10.3969/j.issn.1005-4847.2015.03.002

Objective To establish a mouse model of early lung adenocarcinoma to serve the imaging studies of early lung adenocarcinoma .Methods Two-hundred and ten 4-week old SPF female Kunming mice were randomly divided into 5 experimental groups (40 mice per group) and 1 control group (10 mice).The mice of experimental groups were subcutaneously injected with a dose of 0.2 ml 1-methyl-3-nitro-1-nitroso-guanidine ( MNNG) solution in concentration of 2.0 mg/mL weekly for 1, 2, 4, 8, 12 weeks (group A-E), respectively, while the mice of control group was subcutane-ously injected with 0.2 mL saline weekly for 12 weeks.Ten mice were randomly sacrificed from each experimental group at the 60th, 80th, 100th and 120th days after initial injection ( group A60 , group A80 , group A100 , group A120; group B60 ,……and so on) , and the bilateral lungs were dissected .The tumor occurrence rate , number and size of lung tumors were observedandrecorded.Results 1.Thetumoroccurrencerateandnumberoflungtumorswerepositivelyproportionalto the time and dose of MNNG injection .No tumor was found in any group at the 60th day.2.At the 80th day, the tumor oc-currence rate in the groups A, B, C, D and E was 0, 10%, 30%, 40%and 50%, respectively.The number of tumors in each group was 0, 4, 42, 60 and 81, respectively.The number of tumors smaller than 0.5 mm in diameter was 0, 4, 25, 31 and 40, respectively.3.At the 100th day, the tumor occurrence rate was 20%, 40%, 100%, 100%and 100%, re-spectively.The number of tumors was 6, 19, 187, 223 and 301, respectively, and the number of tumors smaller than 0.5 mm in diameter was 5, 16, 132, 124 and 123, respectively.4.At the 120th day, the tumor occurrence rate was 30%, 50%, 100%, 100%and 100%, respectively.The number of tumors was 30, 124, 302, 351 and 362 per group, respec-tively.The number of tumors smaller than 0.5 mm in diameter was 21, 98, 123, 140 and 108, respectively.The induced lung adenocarcinomas were confirmed by pathology .Conclusions MNNG solution 0.2 mL (2.0 mg/mL) subcutaneously injected weekly for 4 weeks can produce 100%occurrence rate of lung adenocarcinoma in mice and most tumors are smaller than 0.5 mm in diameter at 100th day after initial injection .It is a most suitable model for imaging studies of mouse early lung adenocarcinoma .

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Interventional effect of cisplatin combined with fluoxetine on mice bearing Lewis lung cancer with depression

Mihui LI ; Xiao WU ; Kai WEI ; Yanhua LIN ; Ying WEI ; Jing SUN ; Qingli LUO ; Baojun LIU ; Jingcheng DONG

Acta Laboratorium Animalis Scientia Sinica.2015;(3):221-226. doi:10.3969/j.issn.1005-4847.2015.03.001

Objective To observe if fluoxetine has a potency to inhibit the progression of Lewis lung cancer by combining the fluoxetine and cisplatin to treat the mice bearing Lewis lung cancer with depression .Methods We devel-oped a mouse model of Lewis lung cancer with depression which was intervened with cisplatin and fluoxetine , and the indi-cators related to cancer and depression were tested .Social interaction test was used to measure the behavioral changes of the depressed model mice .The serum cortisol and IL-6, BDNF mRNA in the hippocampus , and NF-κB and VEGF in the tumor tissue were selected for investigation and comparison .Results The mice which were induced by social defeat exhib-ited social avoidance behavior in the social interaction test .The cortisol and IL-6 level in both combination groups was de-creased compared with that in the model group (P<0.05), and the cortisol and BDNF mRNA in the combination group in-creased signifuicantly ( P <0.05 ) .There were no statistically significant differences in the tumor weight , NF-κB and VEGF in tumor tissues between the single and combination groups (P>0.05).Conclusions Fluoxetine has antidepres-sant effect by decreasing the high level of serum cortisol and promoting the BDNF mRNA expression in the hippocampus . However , fluoxetine is not found to have the potency to inhibit the expression of NF -κB related with the progression of tumor.

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Electroacupuncture promotes revascularization via eNOS mobilizing EPCs in the middle cerebral artery occlusion/reperfusion rat

Yanhan ZHU ; Yong LUO ; Hongbei XU ; Panxin WANG

Acta Laboratorium Animalis Scientia Sinica.2015;(3):291-296. doi:10.3969/j.issn.1005-4847.2015.03.013

Objective To explore the effect and mechanism of electroacupuncture on eNOS in mobilizing endothe -lial progenitor cells (EPCs) in rat bone marrow and peripheral blood to promote revascularization in focal cerebral ischemi -a/reperfusion rat.Methods A total of 100 healthy male adult Sprague-Dawley (SD) rats were randomly divided into nor-mal group ( N) , model group ( I/R) , electroacupuncture group ( I/RE) and I/RE plus L-NAME ( A specific antagonist of eNOS) group (I/REL), and were further divided into 1 d, 2 d and 7 d subgroups after reperfusion, 10 rats in each group, in addition to the N group .The rats received filament occlusion of the right middle cerebral artery for 1.5 hours followed by reperfusion .“Baihui” ( GV 20)/“Siguan” ( Hegu LI 4/Taichong LR 3) were selected as acupucture points .Flow cytome-ter was used to detect the percentage of EPCs in bone marrow and peripheral blood .The expression of VEGFR2 mRNA was tested by fluorescence quantitative PCR .Immunohistochemical staining was used to detect VEGFR 2 +cells and to stain the CD34 +microvessels.Results Compared with the I/R group, there was a significant up-regulation of the percentage of EPCs in bone marrow and peripheral blood by electroacupuncture (P<0.01,P<0.05), but decreased by the inhibitor of eNOS (P<0.01).Compared with the I/R group, the VEGFR2 +cells, expression of VEGFR2 mRNA and CD34 +mi-crovessels were significantly increased in the I/RE group ( P<0.01 ) , but decreased in the I/REL group ( P<0.01 ) . Conclusions Electroacupuncture can effectively mobilize EPCs to promote the revascularization in focal cerebral ischemia /reperfusion rat .This effect is attenuated by inhibitor of eNOS , suggesting that the activation of eNOS mediated by electroa-cupuncture may be related to mobilizing EPCs to promote the revascularization .

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Effect of refraction and axial length on retinal nerve fiber layer thickness and optic disc morphology in guinea pigs

Tao LI ; Xiaodong ZHOU ; Hanran JIANG ; Jie ZHU

Acta Laboratorium Animalis Scientia Sinica.2015;(3):285-290. doi:10.3969/j.issn.1005-4847.2015.03.012

Objective To observe the retinal nerve fiber layer ( RNFL) thickness and optic disc morphology in guinea pigs by optical coherence tomography ( OCT ) , and to investigate the correlation of spherical equivalent and axial length with those parameters .Methods Twenty healthy guinea pigs were used in this study .Spherical equivalent and axi-al length of nine guinea pigs were measured , and retinal nerve fiber layer thickness and optic disc morphology in the guinea pigs were analyzed by OCT .Results Spherical equivalent of the guinea pigs was positively correlated with average RNFL thickness, and with superior, temporal, inferior and nasal RNFL thicknesses;while ocular axial length was negatively cor-related with average RNFL thickness , and with superior , temporal , inferior and nasal RNFL thicknesses .Spherical equiva-lent and axial length did not correlate significantly with disc edge area , optic disc area , average cup disc ratio , or cup vol-ume .Vertical cup disc ratio significantly correlated with axial length , but did not with spherical equivalent .Conclusions The retinal nerve fiber layer thickness in guinea pigs is affected by both spherical equivalent and axial length .When guinea pigs are used as an animal model of glaucoma , the effect of refractive state and axial length should be considered .

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Effects of electroacupuncture on the expressions of IL-1βand ICAM-1 in bilateral brain tissues of rats with cerebral ischemia/reperfusion injury

Yingzhou SONG ; Linlin SUN ; Yingzhen REN ; Xuhui ZHANG ; Mingmin XU ; Miao YU ; Yu GUO ; Ya TU

Acta Laboratorium Animalis Scientia Sinica.2015;(3):278-284. doi:10.3969/j.issn.1005-4847.2015.03.011

Objective To explore the variation trends of interleukin-1β( IL-1β) and intercellular adhesion molecule-1 (ICAM-1) in both normal and affected sides of brain tissues in rats with ischemia-reperfusion injury and the therapeutic action of electroacupuncture.Methods The cerebral ischemia-reperfusion model was established with suture embolization in the right middle cerebral artery.The rats were randomly divided into control group, model group and electroacupunture group.Each group was then divided into six subgroups by the time after operation (12 h,24 h,48 h,72 h,96 h,144 h), ten rats in each subgroup. Frozen sections of brain tissues were prepared and the expression of IL-1βand ICAM-1 in brain tissues of both sides were detec-ted by immunohistochemistry.Results The expressions of IL-1βand ICAM-1 showed typical bimodal pattern in both affected is-chemic region and contralateral normal region.In the model group, the peaks of IL-1βin the cerebral ischemic region were at 12 h and 48 h, while in the contralateral normal region the peaks were at 12 h and 144 h, the expression of IL-1βin the ischemic region was significantly higher than that in the contralateral normal region at 48 h (P<0.05), and lower at 96 h and 144 h (P <0.05).In the electroacupuncture group, the expressions of IL-1βin the ipsilateral region were significantly lower than that in the contralateral region at 24 h, 48 h and 144 h (P<0.05).In the model group, the peaks of ICAM-1 in the cerebral ischemic regions were at 24 h and 72 h, while in the contralateral normal regions the peaks were at 24 h and 144 h.In the electroacupunc-ture group, the expressions of ICAM-1 in the ischemic regions were significantly lower than that in the contralateral normal re-gions at all the 12 h, 24 h, 48 h, 72 h and 144 h (P<0.05).Conclusions Our findings suggest that electroacupuncture may inhibit the inflammation of ischemia/reperfusion brain tissue through reducing the expression of IL-1βand ICAM-1 to relieve the cerebral ischemia-reperfusion injury.

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Mammary dysplasia and abnormal estrogen/progesterone or their receptor expression induced by psychological stress in pregnant rats

Ruiqiong WANG ; Guotai WU ; Fenglin LIU ; Yanming WEI ; Yuhong WU

Acta Laboratorium Animalis Scientia Sinica.2015;(3):272-277. doi:10.3969/j.issn.1005-4847.2015.03.010

Objective To study the effects of psychological stress on breast development and abnormal levels of related hormones or their receptors in pregnant rats .Methods Pregnant Wistar rats were randomly divided into control group and ex-perimental group.The experimental group rats were given unpredictable stress stimuli for 15 days, including noise, restraint, re-versal of day and night , swimming in cold water and tail pinch-induced pain.Gross changes of the breast were observed , the di-ameter and height of breast nipple was measured, and breast coefficient was calculated.DNA and RNA levels in the breast tis-sues were determined by optical density measurement.The estrogen (E2), progesterone (P), growth hormone (GH) and prolac-tin (PRL) levels in plasma and breast tissues were determined by radioimmunoassay.Bmax and Kd of estradiol and progesterone receptors were measured by radolig and bindng assay .Morphological changes of the breast tissues were examined by light micros-copy using HE staining.Results Compared with the control group, the nipple diameter and height, breast weight and breast co-efficient were significantly decreased in the experimental group (P<0.05, P<0.01), the levels of DNA, RNA and RNA/DNA ratio in the breast tissue were significantly decreased (P<0.05, P<0.01), the levels of plasma E2, P and GH were significant-ly decreased (P<0.05, P<0.01), the levels of E2 and GH in the breast tissue homogenate were decreased also (P<0.05, P<0.01), the Bmaxs of E2 receptor and P receptor were decreased, but their Kds increased significantly (P<0.05, P<0.01), and the number of lobules and diameter of acini were significantly decreased (P<0.05).Conclusions Psychological stress can lead to breast dysplasia and abnormal levels of estrogen and progesterone and their receptors in pregnant rats .

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Changes of expression of angiogenic factor mRNA in the spinal cord of rat models of protruded intervertebral disc and the effects of electroacupuncture

Weipeng WU ; Wei LI ; Peng CHENG ; Daixun JIANG ; Wu CHEN

Acta Laboratorium Animalis Scientia Sinica.2015;(3):261-266. doi:10.3969/j.issn.1005-4847.2015.03.008

Objective The purpose of this study was to determine the changes of expression of angiogenic factor mRNA in the spinal cord of rats with protruded intervertebral disc and the effects of electroacupuncture , and to analyze the role of electroacupuncture in the spinal microvascular angiogenesis .Method Eighteen healthy male Sprague-Dawley rats were divided randomly into 3 groups:the electroacupuncture group , model group and sham-operated group .The protruded intervertebral disc models were generated by the left lateral and ventral spinal cord compression at T 13 using a self-made silicon sheet.Only the rats in the first group were treated by electroacupuncture once a day for consecutive 7 days.Spinal cord tissue samples were taken from the compression site at 7 days after operation .The mRNA levels of Ang-1, Tie-1, Ang-2, Tie-2, VEGF, Flt-1, caspase-3 and Tsp-1 were determined by RT-PCR, and the pathological changes of the spinal cord was examined using HE staining .Results The rats in the electroacupuncture treated group showed significantly im-proved hind leg function , a relatively complete spinal cord structure , and a clear boundary between grey and white matters . The expression levels of Ang-1, Ang-2, Tie-1, Tie-2, capase-3 and Tsp-1 in the spinal cord tissues of the model group were significantly higher than those of sham-operated group ( P<0.01 ) , but the levels of VEGF and Flt-1 between those two groups showed no significant difference (P>0.05).The expression levels of Ang-2, Tie-1, Tie-2, caspase-3 and Tsp-1 of the electroacupuncture group were significantly lower than those in the model group (P<0.01), while the expression levels of Ang-1, VEGF and Flt-1 were not significantly different between the two groups (P>0.05).All the indexes be-tween electroacupuncture and sham-operated groups showed no significant difference ( P>0.05 ) .Conclusions Our re-sults demonstrate that mRNA expression of relevant angiogenic factors were abnormal after spinal cord compression , while electroacupuncture can down-regulate the expression of Ang-2, Tie-2,Tsp-1 and caspase-3, and modulate the promoting and inhibiting factors of angiogenesis to return towards normal , therefore , to create beneficial conditions for the repair of spinal cord injuries .

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