CAJ | 학술논문

目的通过检测内毒素血症模型动物骨骼相关基因表达变化,进一步明确内毒素对机体的损害机制,为下一步研究骨骼变化对脏器产生何种影响奠定基础.方法腹腔注射内毒素脂多糖(LPS)制作内毒素血症小鼠模型.将48只小鼠随机分为正常组和LPS处理4、6、8、12、24、48和72 h组,每组6只.采尾血计数全血白细胞;取眼眶血检测肝、肾功能;用逆转录-聚合酶链反应(RT-PCR)检测骨骼中TLR4 mRNA表达;苏木素-伊红(HE)染色,镜下观察骨骼病理学变化.结果与正常组比较,LPS 4 h组白细胞计数显著下降(P＜0.01),之后逐渐升高,于72 h时显著高于正常组(P＜0.05);LPS 4 h和6 h组丙氨酸转氨酶(ALT)水平显著升高(P均＜0.05),于8 h降至正常水平(P＞0.05);LPS 6 h组血尿素氮(BUN)水平逐渐升高(P＜0.05),于8 h达峰值(P＜0.05),然后下降,12 h趋于正常(P＞0.05);LPS 6 h组TLR4 mRNA表达增加(P＜0.01),24 h达峰值(P＜0.01),然后逐渐下降,72 h仍处在较高水平(P＜0.05).LPS作用于骨骼后,HE染色显示骨骼无明显改变.结论内毒素血症中骨骼TLR4 mRNA表达量显著增加.
목적 통과검측내독소혈증모형동물골격상관기인표체변화,진일보명학내독소대궤체적손해궤제,위하일보연구골격변화대장기산생하충영향전정기출.방법 복강주사내독소지다당(LPS)제작내독소혈증소서모형.장48지소서수궤분위정상조화LPS처리4、6、8、12、24、48화72 h조,매조6지.채미혈계수전혈백세포;취안광혈검측간、신공능;용역전록-취합매련반응(RT-PCR)검측골격중TLR4 mRNA표체;소목소-이홍(HE)염색,경하관찰골격병이학변화.결과 여정상조비교,LPS 4 h조백세포계수현저하강(P＜0.01),지후축점승고,우72 h시현저고우정상조(P＜0.05);LPS 4 h화6 h조병안산전안매(ALT)수평현저승고(P균＜0.05),우8 h강지정상수평(P＞0.05);LPS 6 h조혈뇨소담(BUN)수평축점승고(P＜0.05),우8 h체봉치(P＜0.05),연후하강,12 h추우정상(P＞0.05);LPS 6 h조TLR4 mRNA표체증가(P＜0.01),24 h체봉치(P＜0.01),연후축점하강,72 h잉처재교고수평(P＜0.05).LPS작용우골격후,HE염색현시골격무명현개변.결론 내독소혈증중골격TLR4 mRNA표체량현저증가.
Objective To explore the changes in gene expression of bone in endotoxemia in mice.Methods Lipopolysaccharide(LPS)was injected intraperitoneally to reproduce an endotoxemia model in mice.Forty-eight mice were randomly divided into eight groups:normal group,4,6,8,1 2,24,48 and 72 hours after LPS injection groups,with 6 mice in each group.To evaluate endotoxemia whole blood was collected for leukocytic count,eye sockets blood was obtained for liver and renal functions tests.mRNA expression level of Toll-like receptor 4(TLR4)in bone was determined by reverse transcription-polymerase chain reaction(RT-PCR)assay.The pathological changes of bone and tissue slides were prepared with hematoxylin and eosin(HE)staining for observing under microscope.Results Compared with normal group,leukocyte count of 4-hour LPS-treated group was significantly decreased(P<0.01).However.after 4 hours,leukocyte count became higher gradually and remained at high level at 72 hours compared with that of normal group(P<0.05).Compared with normal group,alanine aminotransferase(ALT)level reached to a high level in LPS-treated groups at 4 hours and 6 hours(both P<0.05),and then decreased gradually showing a tendency to return to normal level after 8 hours(P>0.05).Blood urea nitrogen(BUN) was increased at 6 hours(P<0.05),reaching to the highest level at 8 hours(P<0.05)and tended to become normallevel after 12 hours(P>0.05).Six hours after LPS treatment,the expression of TLR4 mRNA was enhanced(P<0.01)and reached the peak at 24 hours(P<0.01),and it remained at a high level at 72 hours (P<0.05).However,there were no significant pathological changes in bone structure after LPS treatment.