中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2014年
12期
814-819
,共6页
氟%信号传导%药物性肝损伤,慢性%疾病模型,动物
氟%信號傳導%藥物性肝損傷,慢性%疾病模型,動物
불%신호전도%약물성간손상,만성%질병모형,동물
Fluorine%Signal transduction%Drug-induced liver injury,chronic%Disease models,animal
目的：研究sonic hedgehog（ Shh）信号通路在慢性氟中毒大鼠肝脏组织中的表达情况以及环杷明（ cyclopamine）的抑制作用，探讨Shh信号通路对慢性氟中毒大鼠肝脏损伤作用的机制及其意义。方法建立慢性氟中毒大鼠动物模型，选取4～5周龄，健康SD大鼠48只，饲养1周后按体质量均衡分为4组，每组12只，正常组、氟中毒组、阻断剂组以及阻断剂对照组。饲养6个月后阻断剂组腹腔注射环杷明，阻断剂对照组腹腔注射二甲基亚砜，10 mg／kg，隔天1次，注射3次。收集大鼠24 h尿液检测尿氟后处死大鼠，应用自动血生化分析仪检测大鼠肝功能，通过免疫组织化学（ IHC ）、Western blot和即时荧光定量PCR （ real－time PCR ）的方法检测大鼠肝脏组织中 Shh、Gli1蛋白和mRNA的表达。结果（1）与对照组相比，染氟组大鼠氟斑牙发生率明显升高，各组尿氟含量均增加。（2）肝功能检查显示，与对照组相比，染氟组的肝脏功能明显地改变，氟中毒组、阻断剂对照组中丙氨酸转氨酶和天冬氨酸转氨酶活性升高，而总蛋白和白蛋白含量降低；与染氟组比较，阻断剂组丙氨酸转氨酶活性降低，而天冬氨酸转氨酶、总蛋白和白蛋白含量升高。（3）肝形态学改变：光镜下观察发现，与对照组相比，实验组大鼠肝脏肝细胞索排列紊乱，肝细胞肿大，部分肝细胞胞质透亮，呈气球样改变。（4）与对照组相比，氟中毒组、阻断剂对照组中Shh、Gli1蛋白和mRNA的表达升高，阻断剂组较氟中毒组、阻断剂对照组降低。结论过量氟可以激活慢性氟中毒大鼠肝组织中Shh信号通路，环杷明能有效地抑制Shh信号通路激活，故Shh信号通路可能参与慢性氟中毒引起肝脏损伤发病的过程，研究该通路的作用机制对慢性氟中毒肝脏损伤的预防、诊疗有较大的意义。
目的：研究sonic hedgehog（ Shh）信號通路在慢性氟中毒大鼠肝髒組織中的錶達情況以及環杷明（ cyclopamine）的抑製作用，探討Shh信號通路對慢性氟中毒大鼠肝髒損傷作用的機製及其意義。方法建立慢性氟中毒大鼠動物模型，選取4～5週齡，健康SD大鼠48隻，飼養1週後按體質量均衡分為4組，每組12隻，正常組、氟中毒組、阻斷劑組以及阻斷劑對照組。飼養6箇月後阻斷劑組腹腔註射環杷明，阻斷劑對照組腹腔註射二甲基亞砜，10 mg／kg，隔天1次，註射3次。收集大鼠24 h尿液檢測尿氟後處死大鼠，應用自動血生化分析儀檢測大鼠肝功能，通過免疫組織化學（ IHC ）、Western blot和即時熒光定量PCR （ real－time PCR ）的方法檢測大鼠肝髒組織中 Shh、Gli1蛋白和mRNA的錶達。結果（1）與對照組相比，染氟組大鼠氟斑牙髮生率明顯升高，各組尿氟含量均增加。（2）肝功能檢查顯示，與對照組相比，染氟組的肝髒功能明顯地改變，氟中毒組、阻斷劑對照組中丙氨痠轉氨酶和天鼕氨痠轉氨酶活性升高，而總蛋白和白蛋白含量降低；與染氟組比較，阻斷劑組丙氨痠轉氨酶活性降低，而天鼕氨痠轉氨酶、總蛋白和白蛋白含量升高。（3）肝形態學改變：光鏡下觀察髮現，與對照組相比，實驗組大鼠肝髒肝細胞索排列紊亂，肝細胞腫大，部分肝細胞胞質透亮，呈氣毬樣改變。（4）與對照組相比，氟中毒組、阻斷劑對照組中Shh、Gli1蛋白和mRNA的錶達升高，阻斷劑組較氟中毒組、阻斷劑對照組降低。結論過量氟可以激活慢性氟中毒大鼠肝組織中Shh信號通路，環杷明能有效地抑製Shh信號通路激活，故Shh信號通路可能參與慢性氟中毒引起肝髒損傷髮病的過程，研究該通路的作用機製對慢性氟中毒肝髒損傷的預防、診療有較大的意義。
목적：연구sonic hedgehog（ Shh）신호통로재만성불중독대서간장조직중적표체정황이급배파명（ cyclopamine）적억제작용，탐토Shh신호통로대만성불중독대서간장손상작용적궤제급기의의。방법건립만성불중독대서동물모형，선취4～5주령，건강SD대서48지，사양1주후안체질량균형분위4조，매조12지，정상조、불중독조、조단제조이급조단제대조조。사양6개월후조단제조복강주사배파명，조단제대조조복강주사이갑기아풍，10 mg／kg，격천1차，주사3차。수집대서24 h뇨액검측뇨불후처사대서，응용자동혈생화분석의검측대서간공능，통과면역조직화학（ IHC ）、Western blot화즉시형광정량PCR （ real－time PCR ）적방법검측대서간장조직중 Shh、Gli1단백화mRNA적표체。결과（1）여대조조상비，염불조대서불반아발생솔명현승고，각조뇨불함량균증가。（2）간공능검사현시，여대조조상비，염불조적간장공능명현지개변，불중독조、조단제대조조중병안산전안매화천동안산전안매활성승고，이총단백화백단백함량강저；여염불조비교，조단제조병안산전안매활성강저，이천동안산전안매、총단백화백단백함량승고。（3）간형태학개변：광경하관찰발현，여대조조상비，실험조대서간장간세포색배렬문란，간세포종대，부분간세포포질투량，정기구양개변。（4）여대조조상비，불중독조、조단제대조조중Shh、Gli1단백화mRNA적표체승고，조단제조교불중독조、조단제대조조강저。결론과량불가이격활만성불중독대서간조직중Shh신호통로，배파명능유효지억제Shh신호통로격활，고Shh신호통로가능삼여만성불중독인기간장손상발병적과정，연구해통로적작용궤제대만성불중독간장손상적예방、진료유교대적의의。
Objective To investigate the expression of sonic hedgehog( Shh) signaling pathway in liver fluorosis and to explore related mechanism.Methods To establish animal model, 48 normal SD rats ( aged 4-5 weeks) were randomly divided into 4 groups(12 each):control group, fluoriosis group, blocking group and blocking control group.After 6 months, the blocking group and blocking control group were injected intraperitoneally once every 2 days for 3 times with 10 mg/kg cyclopamine or dimethysulfoxide, respectively.Rats were sacrificed at the end of the experiment and the fluoride content in urine and liver function was determined.The expression of Shh and Gli1 protein and mRNA in hepatocytes was detected by immunohistochemistry and real-time fluorescence quantitative PCR, respectively.Results The fluoride contents in the urine and the incidence of dental fluorosis increased in the fluoride and blocking control groups as compared with those in the control group, but decreased in the blocking group compared with those of the fluoride and blocking control group. Compared with the control group, the titers of aspartate transaminase (AST) and alanine transaminase ( ALT) significantly increased, while the activity of total protein and albumin decreased in the fluoride and blocking control groups.Compared with the fluoride and blocking control groups, the activity of the ALT slightly declined and the AST, total protein and albumin slightly increased in the blocking group. Histologically, the cells were disorganized and swollen with cytoplasmic clearing ( balloon cells ) , compared with the control group.The expression of Shh and Gli1 significantly increased in all but the control group.Compared with the fluoride and blocking control groups, the expression of Shh and Gli1 declined in the blocking group.Conclusions The overexpression and cyclopamine inhibition of the Shh signaling pathway are closely related to the content of fluoride in the liver. The Shh signaling pathway plays an important role in the pathogenesis of liver injury caused by fluorosis, suggesting a preventive and therapeutic target of the disease.
