Sex differences in morphine-induced behavioral sensitization and social behaviors in ICR mice

Gender and genetic strain are two prominent variants that influence drug abuse. Although certain sexrelated behavioral responses have been previously characterized in ICR mice, little is known about the effects of sex on morphine-induced behavioral responses in this outbred strain. Therefore, in this study, we investigated the sex differences of morphine-induced locomotion, anxiety-like and social behaviors in ICR mice. After morphine or saline exposure for four consecutive days（twice daily）, increased locomotion, more time spent in the central area, as well as attenuated rearing and self-grooming behaviors were found in morphine-treated females in an open field; no differences were found in locomotion and the time spent in the central area between male and female controls. When interacting with the samesex individuals, female controls were engaged in more social investigation, following, body contacting and self-grooming behaviors than controls; morphine exposure reduced contacting and self-grooming behaviors in females; in contrast, these effects were not found in males. These results indicate that female ICR mice are more prosocial and are more susceptible to morphine exposure than males.

Hematological effects of Ipomoea batatas(camote) and Phyllanthus niruri(sampa-sampalukan) from Philippines in the ICR mice(Mus musculus)

Objective:To analyze the hematological effects of administering Ipomoea batatas(I.batatas)and Phyllanthus niruri(P.niruri) in the ICR mice.Methods:Powdered leaves of /.batatas and P.nintri were fed to mice for 4 weeks.A total of six groups were used to determine the effect of the plants to the complete blood count of the mouse.Group A(blank control) mice were feed with pellets only;Group B(negative control) mice were fed with pellets coated with honey;Group C(low dosage) mice were fed with honey-coated pellets and powdered leaves of 1.batatas at 10 g/kg body weight of the mouse;Group D(high dosage) mice were fed with honey-coated pellets and powdered leaves of I,batatas at 20 g/kg body weight of the mouse;Group E(low dosage) mice were fed with honey-coated pellets and powdered leaves of P.niruri at 10 g/kg body weight of the mouse:and Group F(high dosage) mice were fed with honey-coated pellets and powdered leaves of P.niruri at 20 g/kg body weight of the mouse.Complete blood count was performed on Days 0.14 and 28.Results:It was shown that I.batatas can increase the values of hematocrit and hemoglobin on both the low dose and high dose at Day 28 and red blood cells(RBC) on both Days 14 and28 of testing.On the other hand.P.niruri can increase RBC.hematocrit and hemoglobin on Day 28 with only the low dose.There were no significant differences with white blood cell,absolute granulocyte,lymphocyte and monocyte,and platelet counts observed for both plant samples.Conclusions:I.batatas and P.niruri have effects on the hematocrit,RBC and hemoglobin levels in mice.

Anti-hyperglycemic effects of aqueous Lenzites betulina extracts from the Philippines on the blood glucose levels of the ICR mice(Mus musculus)

Objective: To examine the anti-hyperglycemic effects of aqueous Lenzites betulina(L. betulina) extracts on normoglycemic glucose-loaded mice.Methods: Different doses of aqueous extract from L. betulina were administered to 45 ICR mice(Mus musculus) to determine whether there was an effect of L. betulina extracts on the blood glucose level of the ICR mice. Aqueous extracts of L. betulina were orally gavaged to mice using oral glucose tolerance test. A total of five groups were used to determine the effect of the fungi on blood glucose of the mice. Group A(positive control)was given 16.7 mg/kg glimepiride; Group B(negative control) was given distilled water;Group C(low dosage) was given 200 mg/kg aqueous extract; Group D(mid dosage) was given 400 mg/kg aqueous extract and Group E(high dosage) was given 800 mg/kg aqueous extract. Baseline blood glucose value was firstly acquired before induction of hyperglycemia through D-glucose, after which another check on blood glucose was made after 0.5 h. Immediately, after the acquisition of hyperglycemic blood glucose level, the individual administration of treatments were done. After that, three blood collections were done spanning 3 h with 1 h interval.Results: The low dose(200 mg/kg) and the mid dose(400 mg/kg) of L. betulina extracts were significantly different(P < 0.05) from their respective baseline values throughout the whole experiment with the latter surpassing its baseline value during the 3rd hour. On the other hand, the high dose(800 mg/kg) during the 1st hour after administration was not significantly different(P > 0.05) from its corresponding baseline value, acting faster than the positive control(glimepiride), which only became significantly different(P < 0.05) at the 2nd hour.Conclusions: Aqueous L. betulina extract is able to produce hypoglycemic effects on the mice with all doses, which are able to normalize blood glucose levels at varying times.

Morphine analgesia in male inbred genetic diversity mice recapitulates the among-individual variance in response to morphine in humans

Morphine is a widely used analgesic, but its use in clinical precision medicine is limited by the variance in response among individuals. Although previous studies have shown that individual differences in morphine can be explained in terms of pharmacodynamics and pharmacokinetics, genetic polymorphisms also play an important role. However, the genetic basis of different sensitivity and tolerance susceptibility to morphine remains ambiguous. Using 15 strains of inbred Genetic Diversity(GD) mice,a new resource with wide genetic and phenotypic variation, we demonstrated great variance in sensitivity to morphine analgesia and susceptibility to morphine tolerance between different GD strains. Among-i ndividual variance in response to morphine analgesia in the population can be modeled in GD mice. Two loci respectively may be associated with the among-i ndividual variance in morphine sensitivity and tolerance,confirming the role of genetic factors in among-i ndividual different responses to morphine. These results indicate that GD mice may be a potential tool for the identification of new biomarkers to improve the clinical administration of morphine.

ESTABLISHMENT OF A MAMMARY CANCER CELL LINE Ca 761-86 AND ITS BIOLOGICAL CHARACTERISTICS IN INBRED 615 MICE

A cell line designated as Ca 761-86 has been established from the solid mouse mammary cancer (Ca 761) by suspension culture. It has been passaged for more than 212 generations. Moderate round cells were predominant and most of them were mononuclear. Some characteristics of malignant cells and A-type viral-like particles were observed by electron microscopy. The results of cytochemical studies (DNA, RNA, SDH, 5' AMPase, ACP etc.) were comparable to the ultramicroscopic results. It multiplied approximately 27.4 fold on day 5 with mitotic index reaching 1.8% on day 3. This cell line was a hyperdiploid with karyotype of 45 or 45, -2X, tril2, tri17, +M1-5. Cell agglutination was observed when treated with ConA (≥7 fig, ml). Spontaneous agglutination might also take place without adding any ConA. After 5×106 cells of Ca 761-86 suspension were transplanted into the normal inbred 615 mice by different ways (subcutan eous, intrafoot-pad or intraperitoneal), the transplan lability rate reached 100%. Spontaneous remission was never observed and its metastatic ability reserved. PPLO were not detected. Ca 761-86 may be of value for practical purposes.

Screening for urinary markers predicting hematopoietic stem cell injury induced by busulfan using genetically diverse mice

Background:Busulfan(BU)is an alkylating agent used as a conditioning agent prior to hematopoietic stem cell(HSC)transplantation as it is known to be cytotoxic to host hematopoietic stem and progenitor cells.The susceptibility of HSCs to BU injury plays an important role in the myeloablative efficacy of BU.Different susceptibilities were demonstrated in genetically diverse(GD)mice in our preliminary research.Methods:Three strains of GD mice with different susceptibilities to BU-i nduced HSC injury were used for screening biological markers of HSC injury susceptibility in urine.The urine proteins were analyzed using liquid chromatography coupled with tandem mass spectrometry to screen for differentially expressed proteins.Screening for possible biomarkers based on differences in protein expression abundance was validated using enzyme-l inked immunoassay(ELISA).Results:Functional analysis showed that the differential proteins were all involved in a series of biological pathways related to cellular senescence,apoptosis,and angiogenesis;whereas the differential proteins of the high-susceptible strain were enriched for the regulation of bone marrow microenvironment pathways,those of low-susceptible strain were enriched for the proapoptotic effect of GTPase pathways.Based on protein abundance differences,several urinary proteins that may be indicative of susceptibility were screened,and ELISA validation results showed that angiotensin-converting enzyme may be a potential biomarker predicting HSC susceptibility for BU conditioning.Conclusions:This study indicates that urinary protein levels can reflect differences in susceptibility to BU-i nduced HSC injury.Using GD mice to construct genetic difference models will provide preclinical data for screening BU-related biological markers.

Effects of estradiol-17β and bisphenol A administered chronically to mice throughout pregnancy and lactation on the male pups＇ reproductive system

Aim： To assess the effect of estradiol-17β （E2） and bisphenol A （BPA） administered chronically by implanting a silicone tube throughout pregnancy and lactation on male pups＇ reproductive system in ICR mice. Methods： Female mice were implanted with a tube filled with 10 ng, 500 ng, 1 μg, or 10 μg of E2, or 100 μg or 5 mg of BPA, before mating. The tube was kept in the mice throughout pregnancy and lactation, until the pups had weaned at 4 weeks of age. During the period, E2 was released from the tube at 120 pg or 6, 12 or 120 ng/day, and BPA at 1.2 or 60 μg/day. Results： Most of the mice given 1 μg and 10 lag of E2 did not maintain their pregnancy. However, the other groups showed high rates of birth, more than 70%. At age of 4 weeks, the male pups were killed. Body weight and reproductive organ weights （testes, epididymides and accessory reproductive glands） in the treated groups did not differ from the control values, whereas the percentage of seminiferous tubules in the testis with mature spermatids was significantly lower in the groups given 10 ng and 500 ng of E2 and 5 mg of BPA than that in the control. Conclusion： Chronic exposure to E2 and BPA might disrupt spermatogenesis in male pups. （Asian JAndrol 2008 Mar; 10： 271-276）

Lipofuscin in brains of patients and mice with epidemic hemorrhagic fever

We have previously shown that the lipofuscin in the brain seems to have in-creased in amount in autopsy cases of epidemic hemorrhagic fever.The purpose of thisstudy was to testify if there is really such an increase.Lipfuscin in 10 sections from everybrain of 10 autopsy cases,stained with Sudan Ⅳ,Sudan black and H.E.,was carefully es-timated and found to be greatly increased as compared with the controls of the same agewithout brain disease.Animal experiment was also conducted on 15 sucking BALB/c miceby I.P.inoculation of 100 LD50（0.05ml）of strain Chen of hemorrhagic fever virus,andon 15 mice without inoculation as controls.No lipofuscin was detected in the controls.However,in the brains of experimental mice,lipofuscin was found to be markedly in-creased,especially in the necrotic cells.The findings suggest that the over-productionand deposition of lipofuscin may be a mild change caused by the virus and its related fac-tors,which might be enhanced by hypotension and shock.

Protective effects of cyclosporine A on T-cell dependent ConA-induced liver injury in Kunming mice

INTRODUCTIONThe T-cell dependent specific liver injury in mice induced by concanavalin A(ConA) is a newly cstablished experimental liver injury model,which is considered more eligible for the study on pathophysiology of several human liver discascs,such as viral hepatitis and autommune hepatitis[1-9].T cell activation and several cytokines release had been proven to play a critical role in ConA -induced liver injury[10-19].Cyclosprine A(CsA),an effective inhibitor of activation of T lymphocytc,hes been used widely in clinical treatment,especially in autoimmune diseases and organ transplantation[20-25].In this study,we investigated the possible effect of CsA on ConA-induced liver injury in Kunning mice.

Maternal zinc deficiency impairs brain nestin expression in prenatal and postnatal mice

Effects of maternal dietary zinc deficiency on prenatal and postnatal brain development were investigated in ICR strain mice. From d 1 of pregnancy (EO) until postnatal d 20 (P20), maternal mice were fed experimental diets that contained 1 mg Zn/kg/day (severe zinc deficient, SZD), 5 mg Zn/kg/day (marginal zinc deficient, MZD), 30 mg Zn/kg/day (zinc adequately supplied, ZA) or 100 mg Zn/kg/day (zinc supplemented, ZS and pair-fed, PF). Brains of offspring from these dietary groups were examined at various developmental stages for expression of nestin, an intermediate filament protein found in neural stem cells and young neurons. Immunocytochemistry showed nestin expression in neural tube 10. 5 d post citrus (dpc) as well as in the cerebral cortex and neural tube from 10.5 dpc to postnatal d 10 (P10). Nestin immunoreactivities in both brain and neural tube of those zinc-supplemented control groups (ZA, ZS, PF) were stronger than those in zinc-deficient groups (SZD and MZD). Western blot analysis confirmed that nestin levels in -pooled brain extracts from each of the zinc-supplemented groups (ZA, ZS, PF) were much higher than those from the zinc-deficient groups (SZD and MZD) from 10.5 dpc to P10. Immunostaining and Western blots showed no detectable nestin in any of the experimental and control group brains after P20. These observations of an association between maternal zinc deficiency and decreased nestin protein levels in brains of offspring suggest that zinc deficiency suppresses development of neural stem cells, an effect which may lead to neuroanatomical and behavioral abnormalities in adults.

Molecular mechanism about lymphogenous metastasis of hepatocarcinoma cells in mice

AIM To investigate the correlation between lymphogenous metastasis and matrix metalloproteinases (MMPs) activity and the expression of Fas Iigand of tumor cells in lymph nodes.METHODES Fifty-six inbred 615-mice were equally divided into 2 groups and inoculated with Hca-F and Hca-P cells. Their lymph node metastatic rates were examined.Growth fraction of lymphocytes in host lymph nodes was detected by flow cytometry. The Hca-F and Hca-P cells were cultured with extract of lymph node, liver or spleen.The quantity of MMPs in these supernatants was examined by zymographic analysis. The expression of Fas ligand,PCNA, Bcl-2 protein of Hca-F and Hca-P cells in the mice were examined by immunohistochemistry. The apoptosis signals of macrophages in lymph nodes were observed with in situ DNA fragmentation.RESULTS On the 28th day post-inoculation, the lymph node metastatic rate of Hca-F was 80% (16/20), whereas that of Hca-P was 25% (5/20). The growth fraction of lymphocytes was as follows: in the Hca-F cells, the proliferating peak of lymphocytes appeared on the 14th day post-inoculation and then decreased rapidly, while in HcaP cells, the peak appeared on the 7th day post-inoculation and then kept at a high level. With the extract of lymph node, the quantity of the MMP-9 activity increased (P＜0.01) and active MMP-9 and MMP-2 were produced by both Hca-F and Hca-P tumor cells, which did not produce MMPs without the extract of lymph node or with the extracts of the liver and spleen. The expression of Fas Ligand of Hca-F cells was stronger than that of Hca-P cells (P＜0.01). The expressions of PCNA and Bcl-2 protein of Hca-F cells in the tumors of inoculated area were the same as that of Hca-P cells. In situ DNA fragmentation showed that the positive signals of macrophages were around HcaF cells.CONCLUSION Secretion of MMPs which was associated with metastatic ability of Hca-F and Hca-P tumor cells depends on the environment of lymph nodes. The increased expression of Fas ligand protein of Hca-F tumor cells with high lymphogenous metastatic potential in lymph nodes may help tumor cells escape from being killed by host lymphocytes.

Antihepatoma effect of alpha-fetoprotein antisense phosphorothioate oligodeoxyribonucleotides in vitro and in mice

AIM To evaluate antihepatoma effect ofantisense phosphorothioate oligodeo-xyribonucleotides (S-ODNs) targeted to alpha-fetoprotein (AFP) genes in vitro and in nudemice.METHODS AFP gene expression was examinedby immunocytochemical method or enzyme-linked immunosorbent assay. Effect of S-ODNson SMMC-7721 human hepatoma cell growth invitro was determined using microculturetetrazolium assay. In vivo antitumor activitiesof S-ODNs were monitored by measuring tumorweight differences in treated and control micebearing SMMC-7721 xenografts. Induction of cellapoptosis was evaluated by fluorescence-activated cell sorter (FACS) analysis.RESULTS Antisense S-ODN treatment led toreduced AFP gene expression. Specificantisense S-ODNs, but not control S-ODNs,inhibited the growth of heaptoma cells in vitro.In vivo. only antisense S-ODNs exhibitedobvious antitumor activities. FACS analysisrevealed that the growth inhibition by antisenseS. ODNs was associated with their cell apoptosisinduction.CONCLUSION Antisense S-ODNs targeted toAFP genes inhibit the growth of human hepatomacells and solid hepatoma, which is related totheir cell apoptosis induction.

Purification of heat shock protein 70-associated tumor peptides and their antitumor immunity to hepatoma in mice

AIM:To purify the heat shock protein (HSP) 70-associated tumor peptides and to observe its non-MHC-I molecule restrictive antitumor effect.METHODS:By ConA-sepharose affinity chromatography,ADP-agarose affinity chromatography, and DEAE anion exchange chromatography, we were able to purify HSP70-associated peptides from mouse hepatoma (HCaF) cells treated in heat shock at 42℃. Specific active immunization and adoptive cellular immunization assay were adopted to observe the immunoprotective effect elicited by HSP70-associated peptide complexes isolated from HcaF.RESULTS: The finally purified HSP-associated peptides had a very high purity and specificity found by SDS-PAGE and Western blot. Mice immunized with HSP70-associated peptide complexes purified from HCaF cells were protected from HCaF living cell challenge. This effect was dose dependent.Adoptive immunization of immune spleen cells of mice immunized with HSP70-associated peptide complexes could elicit immunity against HCaF challenge, and the tumor-free mice could resist repeated challenges. This effect could be continuously enhanced by repeated challenge with HCaF living cells. The tumor-free mice could tolerate the challenge for as high as 1×10^7 HCaF cells. The mice immunized once with spleen cells pulsed with HSP70-associated peptide complexes in vitro could also result in a certain adoptive immunity against HCaF.CONCLUSION:High purity and specificity of HSP70-associated peptides could be achieved from tumor cells by the low-pressure affinity chromatography method used in this study. HSP70-associated peptide complexes derived from the HCaF can elicit non-MHC-I molecule restrictive immunoprotective effect against HCaF.This effect can be transferred by adoptive immunization to mice and enhanced by repeated challenge with HCaF live cells.

Hepatitis E virus chimeric DNA vaccine elicits immunologic response in mice

AIM: To construct the plasmid pcHEV23 containing fragments of HEV ORF2 and ORF3 chimeric gene and to assess its ability to elicit specific immunologic response in mice.METHODS: The gene encoding the structural protein of HEV ORF2 fragment and full-length ORF3 was amplified by PCR. The PCR products were cloned into an eucaryotic expression plasmid pcDNA3. The resulting plasmid pcHEV23 was used as a DNA vaccine to inoculate BALB/c mice intramuscularly thrice at a dose of 100 or 200 μg.Mice injected with empty pcDNA3 DNA or saline served as control and then specific immune responses in the mice were detected.RESULTS: After 2-3 times of inoculation, all mice injected with pcHEV23 had anti-HEV IgG seroconversion and specific T lymphocyte proliferation. The lymphocyte stimulation index in the group immunized with pcHEV23(3.1±0.49) was higher than that in the control group (0.787±0.12, P＜0.01). None in the control group had a detectable level of anti-HEV IgG.CONCLUSION: DNA vaccine containing HEV ORF2 and ORF3 chimeric gene can successfully induce specific humoral and cellular immune response in mice.

The Regulatory Action of the Modified Yu Ping Feng Tang on Cellular Immunity in Mice under Amputation-Induced Stress

To approach the action of modified Yu Ping Feng Tang(玉屏风汤 Jade-Screen Decoction)on cellularimmunity,an experiment was conducted in mice under amputation-induced stress.On the 3rd dayafter amputation,acute atrophy was found in the thymus,the reactivities of T-and B-lymphocytes toCon-A and LPS were decreased,the IL-2 content and its activity reduced and the activity of NK cellslowered.The high,moderate and low concentrations of the modified Yu Ping Feng(YPF)Decoction allhave antagonistic action on the above manifestations of immune inhibition.

ICR鼠肝和肾毒性损伤生物标志物的筛选

肝脏和肾脏是生物体受到外界毒性刺激最易损伤的器官,ICR小鼠又是动物乃至人类病理学评估的重要模式生物。为筛选可用于ICR鼠肝和肾毒性损伤的生物标志物,采用来自受污染的北方两个不同矿区水样对ICR小鼠进行灌胃,建立小鼠肝、肾毒性损伤模型,将其样本进行高通量测序,利用转录组数据进行收集、分组并对比分析,同时采用体外肾细胞NRK的毒性试验进行验证,最终筛选可用于毒性损伤的分子标记物。结果表明,基因Chordc1与Cry1可作为ICR鼠肝和肾毒性损伤生物标志物。试验结果可为有毒有害物对于生物体肝、肾损伤的诊断和健康评价提供一种新思路。

黄精基酒对ICR小鼠抗氧化及抗疲劳作用

为研究黄精基酒对ICR雄性小鼠抗氧化和抗疲劳作用,采用乙醇浸提法提取黄精中功效成分,加入食用乙醇配制出黄精基酒,将60只幼龄ICR雄性小鼠随机分为6组,进行抗氧化实验,分为空白组、抗坏血酸阳性组(120 mg/kg)、白酒阴性组和黄精基酒高、中、低剂量组(33.4、16.7、8.4 mL/kg);另180只小鼠进行黄精基酒抗疲劳实验研究,分为空白组、红景天苷对照组(120 mg/kg)、白酒阴性组和黄精基酒高、中、低剂量组(33.4、16.7、8.4 mL/kg)。通过测定各组小鼠体质量状况,肝肾组织的丙二醛含量、谷胱甘肽含量、过氧化氢酶和超氧化物歧化酶的活性,评价黄精基酒对正常小鼠机体抗氧化效果的影响;通过负重游泳实验,记录各组小鼠的负重游泳时间,小鼠肝糖原、全血乳酸含量和谷丙转氨酶的活性,评价黄精基酒对小鼠机体抗疲劳效果的影响。结果表明,黄精基酒能够有效减少机体内丙二醛含量,增强超氧化物歧化酶和过氧化氢酶的活性,提升谷胱甘肽的含量(P<0.05),并能显著延长小鼠的负重游泳时间(P<0.05),提高小鼠肝糖原含量,降低血乳酸含量和谷丙转氨酶活性(P<0.05)。综上,黄精基酒具有缓解疲劳的作用,能够提高小鼠自身抗氧化能力。

冷应激复温对ICR小鼠神经行为学的影响

【目的】研究不同冷应激复温后对ICR小鼠的自发运动、探索行为、焦虑情绪和血液激素指标的影响。【方法】通过与28℃常温对照组相比,检测ICR小鼠4、10、16、22℃冷应激复温组于复温2、4、6、8、10、12 h后在旷场试验和高架十字迷宫试验中的运动变化,并通过ELISA检测各组在复温2及4 h后的血液激素指标变化。【结果】旷场试验结果显示,与28℃常温对照组相比,22℃冷刺激复温10 h时中央区域停留时间和运动时间显著减少(P<0.05);16℃冷刺激复温4和8 h时中央区域停留时间显著减少(P<0.05);10℃冷刺激复温4和10 h时中央区域停留时间显著减少(P<0.05),复温2 h中央区域运动距离显著增加(P<0.05);4℃冷刺激复温6 h时中央区域停留时间极显著减少(P<0.01),复温2 h中央区域运动距离极显著增加(P<0.01)。高架十字迷宫试验结果显示,与28℃常温对照组相比,22和4℃冷刺激复温4 h、16℃冷刺激复温10 h闭臂停留时间显著或极显著下降(P<0.05;P<0.01);10℃冷刺激复温组闭臂停留时间无显著变化(P>0.05)。与28℃常温对照组相比,皮质酮(CORT)在22℃冷刺激复温4 h时极显著升高(P<0.01),16℃冷刺激复温2和4 h时极显著升高(P<0.01);肾上腺素(EPI)在22℃冷刺激复温4 h时极显著升高(P<0.01),10℃冷刺激复温2和4 h时极显著升高(P<0.01)。【结论】冷应激结束后对ICR小鼠的自发运动、探索行为、焦虑情绪和血液激素指标存在持续影响,应减少以ICR小鼠为模型的试验环境温度变化,提高试验结果准确性。

Effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD_(34)^+ hematopoietic cells in bone marrow of asthmatic mice

BACKGROUND: Corticosteroids remain the most effective therapy available for asthma. They have widespread effects on asthmatic airway inflammation. However, little is known about the effects of corticosteroids on the production of bone marrow inflammatory cells in asthma. This study observed the effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells, so as to explore the possible effectiveness of a bone marrow-targeted anti-inflammatory strategy. METHODS: Balb/c mice were sensitized and challenged with ovalbumin (OVA) to establish an asthmatic model. For two consecutive weeks, asthmatic mice were challenged with OVA while being given either prednisone, montelukast, prednisone plus montelukast, or sterile saline solution. The mice were killed 24 hours after the last challenge with OVA, and bronchoalveolar lavage fluid (BALF), peripheral blood, and bone marrow were collected. Eosinophils in peripheral blood and BALF, and nucleated cells in BALF, peripheral blood, and bone marrow were counted. The percentages of CD34+ cells, CD4+ T lymphocytes and CD8+ T lymphocytes among nucleated cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect expression of CD34 and interleukin (IL)-5Ralpha mRNA (CD34+ IL-5Ralpha mRNA+ cells) among bone marrow hematopoietic cells. RESULTS: Compared with the sterile saline solution group, the number of eosinophils in BALF and peripheral blood, CD34+ cells in peripheral blood and bone marrow, and CD34+ IL-5Ralpha mRNA+ cells in bone marrow of mice from the prednisone and prednisone plus montelukast groups were significantly lower (P

清解化攻方调控NLRP3/TLR4/NF-κB信号通路对重症急性胰腺炎小鼠模型胰腺组织的保护作用

目的观察清解化攻方对重症急性胰腺炎(SAP)小鼠模型的治疗作用,探索清解化攻方抗炎症反应的作用机制。方法将36只C57BL/6J雄性小鼠随机分成空白组,模型组,清解化攻方低、中、高剂量组,西药组(乌司他丁),每组6只,除空白组小鼠,余各组小鼠采用逆行胰胆管注射5%牛黄胆酸钠建立SAP模型,清解化攻方低、中、高剂量组在造模后分别予以清解化攻方1、2、4 g/kg灌胃,西药组在造模后予以腹腔注射乌司他丁(5×10^(4) U/kg),共干预7 d。采用苏木素-伊红染色观察胰腺组织病理改变;酶联免疫吸附测定法(ELISA)检测小鼠α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α水平;RTqPCR检测胰腺组织NOD样受体蛋白3(NLRP3)、Toll样受体4(TLR4)、核因子-κB(NF-κB)mRNA表达水平;免疫组化检测胰腺组织NLRP3、TLR4、NF-κB的阳性表达率;Western Blot技术检测NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白的表达水平。计量资料多组间比较采用方差分析,进一步两两比较采用LSD-t检验。结果与空白组相比,模型组小鼠胰腺组织结构弥漫性破坏、胰腺小叶间隔局灶性扩张、腺泡萎缩和大量炎症细胞浸润,α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α含量明显升高(P值均<0.05),NLRP3、TLR4、NF-κB mRNA表达水平及阳性表达率均明显上升(P值均<0.05),NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白表达均明显上调(P值均<0.05)。与模型组相比,清解化攻方各剂量组和西药组可见小鼠胰腺组织结构稍紧密、完整,胰腺腺泡细胞排列有序,伴少量炎症细胞浸润和胰腺小叶出血灶,α-淀粉酶、脂肪酶、IL-1β、IL-6、IL-8、IL-18和TNF-α含量明显下降(P值均<0.05),NLRP3、TLR4、NF-κB mRNA表达水平及阳性表达率均明显降低(P值均<0.05),NLRP3、TLR4、NF-κB、IL-1β、IL-6蛋白表达水平均明显减弱(P值均<0.05)。结论清解化攻方可能通过抑制NLRP3/TLR4/NF-κB信号通路相关蛋白的激活,减少炎症介质的释放,防止炎症级联反应增强,进而对SAP小鼠胰腺组织发挥保护作用。