中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2015年
4期
238-242
,共5页
徐焰%徐修礼%马越云%沈学锋%刘明朝%任丽芬%张志平%秦庆%邦晓柯
徐燄%徐脩禮%馬越雲%瀋學鋒%劉明朝%任麗芬%張誌平%秦慶%邦曉柯
서염%서수례%마월운%침학봉%류명조%임려분%장지평%진경%방효가
侏儒症,垂体性%铅%铅中毒%生长激素%胰岛素样生长因子Ⅰ%疾病模型,动物
侏儒癥,垂體性%鉛%鉛中毒%生長激素%胰島素樣生長因子Ⅰ%疾病模型,動物
주유증,수체성%연%연중독%생장격소%이도소양생장인자Ⅰ%질병모형,동물
Dwarfism,pituitary%Lead%Lead poisoning%Growth hormone%Insulin-like growth factor I%Disease models,animal
目的 通过生长激素缺乏症(GHD)儿童矮小发生与体内胰岛素样生长因子1(IGF-1)水平、血铅含量之间的相关性研究,以及铅暴露大鼠模型IGF-1相关信号分子的改变,为阐明铅诱导儿童矮小的发生及其致病机制研究提供科学依据,为铅暴露致儿童矮小的监测、防治提供可行途径.方法 采用临床病例对照研究.自2011年6月至2013年3月依据相应诊断标准纳入880例西京医院儿科生长发育门诊就诊矮小患儿,通过生长激素(GH)激发试验GH峰值将患儿分为GHD和特发性矮小(ISS)组,检测并比较不同组别、性别患儿血清IGF-1和血铅水平;醋酸铅饮水法建立生长发育期大鼠铅中毒模型,利用原子吸收分光光度法测定血铅含量确定建模成功;利用蛋白印迹(Western blot)分析确证染铅大鼠脑组织IGF-1相关信号通路分子促分裂原活化蛋白激酶(MAPK)、磷脂酰肌醇-3-激酶/蛋白激酶B(PI3 K/AKT)的变化.采用均值t检验和非参数Mann-Whitney检验等方法进行统计学分析.结果 880例矮小患儿中,GHD患儿362例(41.14％).GHD组患儿IGF-1水平(20.02±11.24) μmol/L(女),(20.74±13.39) μmol/L(男)均低于ISS组患儿IGF-1水平(46.58±27.00) μmol/L(女)、(35.91 ±20.05) μmol/L(男),t分别为10.45和9.98,P均＜0.01.GHD组血铅水平(0.49 ±0.18) μmol/L(女),(0.46 ±0.18) μmol/L(男)高于ISS组血铅(0.32 ±0.11) μmol/L(女),(0.34 ±0.13) μmol/L(男),t分别为-10.91和-9.056,P均＜0.01.铅暴露后大鼠血铅含量逐渐增加,染铅6周时趋于稳定.铅暴露导致信号通路MAPK、AKT中信号分子细胞外信号调节激酶(ERK1/2)、c-Jun氨基末端激酶(JNK)、p38丝裂原活化蛋白激酶(p38)、AKT473、AKT308磷酸化较总酶的蛋白相对表达量增加显著,AKT308/t-AKT组中对照与300 ppm(1 ppm醋酸铅=2.64×10-6 mol/L组蛋白相对表达量分别为1.320±0.071,2.960±0.552,F=19.360,P＜0.01;AKT473/t-AKT 组中对照与300 ppm组蛋白相对表达量分别为0.311 ±0.038,1.018 ±0.282,F=16.101,P＜0.01;p-ERK/t-ERK组中对照与300 ppm组蛋白相对表达量分别为1.173±0.109,6.438±0.748,F=72.054,P＜0.01;p-JNK/t-JNK组中对照与300 ppm组蛋白相对表达量分别为1.249±0.129,4.869±0.907,F=35.528,P＜0.01;p-P38/t-P38组中对照与300 ppm组蛋白相对表达量分别为0.083±0.022,0.500±0.038,F=57.29,P＜0.01.结论 GHD矮小儿童IGF-1水平减低可能与血铅水平升高相关,铅可能通过增加MAPK和AKT等IGF-1相关信号通路分子的磷酸化从而影响IGF-1介导的GH促生长作用,进而导致儿童GHD及矮小的发生.
目的 通過生長激素缺乏癥(GHD)兒童矮小髮生與體內胰島素樣生長因子1(IGF-1)水平、血鉛含量之間的相關性研究,以及鉛暴露大鼠模型IGF-1相關信號分子的改變,為闡明鉛誘導兒童矮小的髮生及其緻病機製研究提供科學依據,為鉛暴露緻兒童矮小的鑑測、防治提供可行途徑.方法 採用臨床病例對照研究.自2011年6月至2013年3月依據相應診斷標準納入880例西京醫院兒科生長髮育門診就診矮小患兒,通過生長激素(GH)激髮試驗GH峰值將患兒分為GHD和特髮性矮小(ISS)組,檢測併比較不同組彆、性彆患兒血清IGF-1和血鉛水平;醋痠鉛飲水法建立生長髮育期大鼠鉛中毒模型,利用原子吸收分光光度法測定血鉛含量確定建模成功;利用蛋白印跡(Western blot)分析確證染鉛大鼠腦組織IGF-1相關信號通路分子促分裂原活化蛋白激酶(MAPK)、燐脂酰肌醇-3-激酶/蛋白激酶B(PI3 K/AKT)的變化.採用均值t檢驗和非參數Mann-Whitney檢驗等方法進行統計學分析.結果 880例矮小患兒中,GHD患兒362例(41.14％).GHD組患兒IGF-1水平(20.02±11.24) μmol/L(女),(20.74±13.39) μmol/L(男)均低于ISS組患兒IGF-1水平(46.58±27.00) μmol/L(女)、(35.91 ±20.05) μmol/L(男),t分彆為10.45和9.98,P均＜0.01.GHD組血鉛水平(0.49 ±0.18) μmol/L(女),(0.46 ±0.18) μmol/L(男)高于ISS組血鉛(0.32 ±0.11) μmol/L(女),(0.34 ±0.13) μmol/L(男),t分彆為-10.91和-9.056,P均＜0.01.鉛暴露後大鼠血鉛含量逐漸增加,染鉛6週時趨于穩定.鉛暴露導緻信號通路MAPK、AKT中信號分子細胞外信號調節激酶(ERK1/2)、c-Jun氨基末耑激酶(JNK)、p38絲裂原活化蛋白激酶(p38)、AKT473、AKT308燐痠化較總酶的蛋白相對錶達量增加顯著,AKT308/t-AKT組中對照與300 ppm(1 ppm醋痠鉛=2.64×10-6 mol/L組蛋白相對錶達量分彆為1.320±0.071,2.960±0.552,F=19.360,P＜0.01;AKT473/t-AKT 組中對照與300 ppm組蛋白相對錶達量分彆為0.311 ±0.038,1.018 ±0.282,F=16.101,P＜0.01;p-ERK/t-ERK組中對照與300 ppm組蛋白相對錶達量分彆為1.173±0.109,6.438±0.748,F=72.054,P＜0.01;p-JNK/t-JNK組中對照與300 ppm組蛋白相對錶達量分彆為1.249±0.129,4.869±0.907,F=35.528,P＜0.01;p-P38/t-P38組中對照與300 ppm組蛋白相對錶達量分彆為0.083±0.022,0.500±0.038,F=57.29,P＜0.01.結論 GHD矮小兒童IGF-1水平減低可能與血鉛水平升高相關,鉛可能通過增加MAPK和AKT等IGF-1相關信號通路分子的燐痠化從而影響IGF-1介導的GH促生長作用,進而導緻兒童GHD及矮小的髮生.
목적 통과생장격소결핍증(GHD)인동왜소발생여체내이도소양생장인자1(IGF-1)수평、혈연함량지간적상관성연구,이급연폭로대서모형IGF-1상관신호분자적개변,위천명연유도인동왜소적발생급기치병궤제연구제공과학의거,위연폭로치인동왜소적감측、방치제공가행도경.방법 채용림상병례대조연구.자2011년6월지2013년3월의거상응진단표준납입880례서경의원인과생장발육문진취진왜소환인,통과생장격소(GH)격발시험GH봉치장환인분위GHD화특발성왜소(ISS)조,검측병비교불동조별、성별환인혈청IGF-1화혈연수평;작산연음수법건립생장발육기대서연중독모형,이용원자흡수분광광도법측정혈연함량학정건모성공;이용단백인적(Western blot)분석학증염연대서뇌조직IGF-1상관신호통로분자촉분렬원활화단백격매(MAPK)、린지선기순-3-격매/단백격매B(PI3 K/AKT)적변화.채용균치t검험화비삼수Mann-Whitney검험등방법진행통계학분석.결과 880례왜소환인중,GHD환인362례(41.14％).GHD조환인IGF-1수평(20.02±11.24) μmol/L(녀),(20.74±13.39) μmol/L(남)균저우ISS조환인IGF-1수평(46.58±27.00) μmol/L(녀)、(35.91 ±20.05) μmol/L(남),t분별위10.45화9.98,P균＜0.01.GHD조혈연수평(0.49 ±0.18) μmol/L(녀),(0.46 ±0.18) μmol/L(남)고우ISS조혈연(0.32 ±0.11) μmol/L(녀),(0.34 ±0.13) μmol/L(남),t분별위-10.91화-9.056,P균＜0.01.연폭로후대서혈연함량축점증가,염연6주시추우은정.연폭로도치신호통로MAPK、AKT중신호분자세포외신호조절격매(ERK1/2)、c-Jun안기말단격매(JNK)、p38사렬원활화단백격매(p38)、AKT473、AKT308린산화교총매적단백상대표체량증가현저,AKT308/t-AKT조중대조여300 ppm(1 ppm작산연=2.64×10-6 mol/L조단백상대표체량분별위1.320±0.071,2.960±0.552,F=19.360,P＜0.01;AKT473/t-AKT 조중대조여300 ppm조단백상대표체량분별위0.311 ±0.038,1.018 ±0.282,F=16.101,P＜0.01;p-ERK/t-ERK조중대조여300 ppm조단백상대표체량분별위1.173±0.109,6.438±0.748,F=72.054,P＜0.01;p-JNK/t-JNK조중대조여300 ppm조단백상대표체량분별위1.249±0.129,4.869±0.907,F=35.528,P＜0.01;p-P38/t-P38조중대조여300 ppm조단백상대표체량분별위0.083±0.022,0.500±0.038,F=57.29,P＜0.01.결론 GHD왜소인동IGF-1수평감저가능여혈연수평승고상관,연가능통과증가MAPK화AKT등IGF-1상관신호통로분자적린산화종이영향IGF-1개도적GH촉생장작용,진이도치인동GHD급왜소적발생.
Objective To investigate the correlation between serum insulin-like growth factor-1 (IGF-1) and blood lead levels in short stature children with growth hormone deficiency (GHD),and study the changes of IGF-1 signal molecules in lead exposed rats,providing evidence for clarifying the pathogenesis of lead induced short stature in children.Methods Totally 800 short stature children were recruited in the clinical case-control study during June 2011 to March 2013 and were divided into GHD group or idiopathic short stature (ISS) group according to the their GH peak in growth hormone stimulation test.The serum IGF-1 levels and blood lead levels were determined.A lead poisoning model in rats was established and Western blot assay was employed to detect the phosphorylation of signaling molecules (MAPK and PI3K/AKT) related to IGF-1 signaling pathway.The average independent samples T-test and non-parametric Mann-Whitney test were used for statistical analysis.Results GHD children accounted for 41.14％ (362 cases) of the 880 short stature cases.Serum IGF-1 levels in GHD group were (20.02 ± 11.24) μmol/L in female and (20.74 ± 13.39) μmol/L in male,which were significantly lower than the ISS Group with (46.58 ±27.00) μmol/L in female and (35.91 ±20.05) μmol/L in male (t =10.45,9.98 respectively,both P ＜0.01.) However,the blood lead levels of GHD group [(0.49 ±0.18) μmol/L in female,(0.46±0.18 μmol/L)] in male were significantly higher than those in ISS group [(0.32 ± 0.11) μmol/L in female,(0.34 ±0.13 μmol/L in male].All had a statistically significant difference (t =-10.91 and-9.056,both P ＜ 0.01).Atomic absorption spectrophotometry showed the blood lead level in rats treated with lead containing water for 6 weeks significantly increased when compared with control group.Western blot assay confirmed that the protein expression of phosphorylated ERK1/2,JNK,p38,AKT473 and AKT308 increased sigrificantly than the total enzyme in lead exposure rats.In AKT308/t-AKT group,the protein expression levels in the control group and 300 ppm(1 ppm =2.64 × 10-6mol/L) group were 1.320 ±0.071 and 2.960 ±0.552(F =19.360,P ＜0.01).In AKT473/t-AKT group,the protein expression levels in the control group and 300 ppm group were 0.311 ± 0.038 and 1.018 ± 0.282,respectively(F =16.101,P ＜0.01).In p-ERK/t-ERK group,the protein expression levels in the control group and 300 ppm group were 1.173 ±0.109 and 6.438 ±0.748(F =72.054,P ＜0.01).In p-JNK/t-JNK group,the protein expression levels in the control group and 300 ppm group were 1.249 ± 0.129 and 4.869 ± 0.907 (F =35.528,P ＜0.01).In p-P38/t-P38 group,the protein expression levels in the control group and 300 ppm group were 0.083 ± 0.022 and 0.500 ± 0.038 (F =57.29,P ＜ 0.01).Conclusions The study suggests that reduction in IGF-1 in children with GHD is associated with an increased blood lead level.Lead possibly affects the IGF-1-mediated growth-promoting effect of GH by increasing the phosphorylation of molecules involved in MAPK,AKT and other IGF-1-related signaling pathways,eventually leading to the occurrence of child GHD and short stature.
