CAJ | 학술논문

目的研究低分子肝素( LMWH)对新生鼠肝细胞先天性感染人巨细胞病毒(HCMV)病毒学和病理学的影响.方法取健康纯系清洁级BALB/c鼠60只,10周龄,雌雄各半,均分为5组,每组6对.LMWH干预组和阳性对照组腹腔接种6.0 lg半数组织培养感染剂量(TCID50)的HCMV AD169,空白对照组腹腔注射高糖改良Eagles培养液0.5 mL/只,然后交配.LMWH干预Ⅰ组的雌鼠在孕期给予皮下注射LMWH 1000 U/kg×10 d;LMWH干预Ⅱ组的雌鼠在孕期给予皮下注射LMWH 1000 U/kg×5d,待生产后再给新生鼠皮下注射LMWH 1000 U/kg×5 d;LMWH干预Ⅲ组在生产后予新生鼠皮下注射LMWH 1000 U/kg×10d.产后10 d处死新生鼠,取肝脏,进行病毒分离、干湿质量测定、病理学检查和荧光定量PCR检测.多组间比较采用方差分析.结果阳性对照组新生鼠肝组织悬液中10d可分离到HCMV,而LMWH干预的3组新生鼠需24 d分离到HCMV.阳性对照组新生鼠肝组织有浊肿、空泡变性,核内有嗜碱性包涵体,部分肝细胞坏死,而LMWH干预组肝组织仅有轻度肝细胞浊肿变性,炎性反应轻微,类似空白对照组.LMWH干预组肝组织含水量较阳性对照组明显降低.LMWH干预的Ⅰ、Ⅱ、Ⅲ组50 mg肝组织HCMV DNA载量对数值分别为(3.26±0.43)、(3.26±0.41)和(3.32±0.51) lg拷贝,较阳性对照组的(7.38±0.53) 1g拷贝显著降低(F=314.620,P＜0.01),但LMWH干预组间比较差异无统计学意义(P＞0.05).结论 LMWH在宫内和出生后干预能显著减少肝细胞HCMV复制,减轻肝组织炎性反应.
목적 연구저분자간소( LMWH)대신생서간세포선천성감염인거세포병독(HCMV)병독학화병이학적영향.방법 취건강순계청길급BALB/c서60지,10주령,자웅각반,균분위5조,매조6대.LMWH간예조화양성대조조복강접충6.0 lg반수조직배양감염제량(TCID50)적HCMV AD169,공백대조조복강주사고당개량Eagles배양액0.5 mL/지,연후교배.LMWH간예Ⅰ조적자서재잉기급여피하주사LMWH 1000 U/kg×10 d;LMWH간예Ⅱ조적자서재잉기급여피하주사LMWH 1000 U/kg×5d,대생산후재급신생서피하주사LMWH 1000 U/kg×5 d;LMWH간예Ⅲ조재생산후여신생서피하주사LMWH 1000 U/kg×10d.산후10 d처사신생서,취간장,진행병독분리、간습질량측정、병이학검사화형광정량PCR검측.다조간비교채용방차분석.결과 양성대조조신생서간조직현액중10d가분리도HCMV,이LMWH간예적3조신생서수24 d분리도HCMV.양성대조조신생서간조직유탁종、공포변성,핵내유기감성포함체,부분간세포배사,이LMWH간예조간조직부유경도간세포탁종변성,염성반응경미,유사공백대조조.LMWH간예조간조직함수량교양성대조조명현강저.LMWH간예적Ⅰ、Ⅱ、Ⅲ조50 mg간조직HCMV DNA재량대수치분별위(3.26±0.43)、(3.26±0.41)화(3.32±0.51) lg고패,교양성대조조적(7.38±0.53) 1g고패현저강저(F=314.620,P＜0.01),단LMWH간예조간비교차이무통계학의의(P＞0.05).결론 LMWH재궁내화출생후간예능현저감소간세포HCMV복제,감경간조직염성반응.
Objective To explore the effects of low molecular weight heparin (LMWH) on the virological and pathological changes of newborn mouse liver congenitally infected with human cytomegalovirus (HCMV).Methods Sixty healthy pure line clean level BALB/c mice which were about 10 weeks old (half were female) were divided into five groups (six pairs in each group).The mice in LMWH intervention group and positive control group were intraperitoneally inoculated with 6.0 lg tissue culture infective dose50 (TCID50) of HCMV AD169; those in blank control group were intraperitoneally injected with 0.5 mL dulbecco's modified Eagles medium (DMEM) ; then all the mice were paired to mate.The pregnant mice in LMWH intervention group Ⅰ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days; those in group Ⅱ were subcutaneously injected with LMWH 1000 U/kg daily for 5 days and their newborn mice were subcutaneously injected with LMWH 1000 U/kg daily for 5 days; those in group Ⅲ were subcutaneously injected with LMWH 1000 U/kg daily for 10 days in their newborn mice.All these newborn mice were sacrificed at day 10 of birth.The liver was removed for virus isolation,dry-wet weight determination,pathology examination and quantitative fluorescence-polymerase chain reaction (PCR) detection.The comparison among groups was done by analysis of variance.Results HCMV was isolated from the supernatant of liver tissue homogenate in 10-day positive control newborn mice,while HCMV was isolated in 24-day newborn mice of the other three groups of LMWH intervention.Pathology confirmed that positive control liver tissue had inflammatory changed,liver cell inflammatory swelling degeneration,vacuoles degeneration,specific HCMV inclusion body in nuclear,and portion of liver cell necrosis,while liver pathological results of LMWH intervention group showed mild liver cell inflammatory changes and slight cell inflammatory swelling degeneration,which were similar to the blank control group.The moisture of liver tissue contents in LMWH intervention group decreased more obviously than positive control group.The HCMV DNA loads in 50 mg liver tissues of LMWH intervention groups Ⅰ,Ⅱand Ⅲ were (3.26±0.43),(3.26±0.41) and (3.32±0.51) lg copy,respectively,which were significantly lower than that of positive control group [(7.38 ± 0.53) lg copy; F =314.620,P0.01],while there were no significant differences among LMWH intervention groups (P＞0.05).Conclusion LMWH intrauterine and postnatal interventions can significantly reduce HCMV DNA replication in hepatocytes,and relieve inflammatory changes in liver tissue.