CAJ | 학술논문

目的研究富含半胱氨酸的酸性分泌蛋白(SPARC)在博莱霉素(BLM)诱导小鼠肺纤维化肺组织中的动态表达及法舒地尔的干预效应,探讨SPARC在肺纤维化中的作用机制.方法将108只小鼠分成正常对照组(Cont组),BLM组,法舒地尔(FAS)组.分别给予等量的0.9％氯化钠注射液0.04ml和BLM(3.5 mg/kg)一次性气管内灌注,FAS组在BLM一次性气管内灌注后给予及FAS 15 mg/kg腹腔注射.各组分别于第3天、第7天、第14天和第28天各处死8只小鼠,取肺组织做病理切片,用苏木精-伊红(HE)染色观察小鼠肺组织的病理形态改变;TUNEL细胞凋亡原位检测BLM组与Cont组肺组织细胞凋亡水平;Western blot检测三组肺组织SPARC及Rho激酶1(Rock-1)蛋白的定量表达,并计算SPARC蛋白与凋亡指数的相关系数.结果小鼠108只进入结果分析,模型组小鼠死亡8只.①HE染色:BLM组肺组织逐渐由肺泡炎进展为纤维化.②TUNEL细胞凋亡原位检测:与Cont组相比,BLM组肺泡上皮细胞凋亡指数呈进行性上升.③Western blot结果显示:BLM组SPARC、Rock-1的表达显著增高.FAS组SPARC、Rock-1的表达逐步下降.BLM组SPARC蛋白与凋亡指数相关系数表明SPARC蛋白表达与肺泡上皮细胞凋亡呈正相关.结论在小鼠肺纤维化进程中存在SPARC过表达,过表达的SPARC蛋白对肺纤维化可能起促进作用.抑制Rho/Rho激酶信号通路可减少SPARC蛋白表达,在延缓肺纤维化进程中可能起重要作用.
목적 연구부함반광안산적산성분비단백(SPARC)재박래매소(BLM)유도소서폐섬유화폐조직중적동태표체급법서지이적간예효응,탐토SPARC재폐섬유화중적작용궤제.방법 장108지소서분성정상대조조(Cont조),BLM조,법서지이(FAS)조.분별급여등량적0.9％록화납주사액0.04ml화BLM(3.5 mg/kg)일차성기관내관주,FAS조재BLM일차성기관내관주후급여급FAS 15 mg/kg복강주사.각조분별우제3천、제7천、제14천화제28천각처사8지소서,취폐조직주병리절편,용소목정-이홍(HE)염색관찰소서폐조직적병리형태개변;TUNEL세포조망원위검측BLM조여Cont조폐조직세포조망수평;Western blot검측삼조폐조직SPARC급Rho격매1(Rock-1)단백적정량표체,병계산SPARC단백여조망지수적상관계수.결과 소서108지진입결과분석,모형조소서사망8지.①HE염색:BLM조폐조직축점유폐포염진전위섬유화.②TUNEL세포조망원위검측:여Cont조상비,BLM조폐포상피세포조망지수정진행성상승.③Western blot결과현시:BLM조SPARC、Rock-1적표체현저증고.FAS조SPARC、Rock-1적표체축보하강.BLM조SPARC단백여조망지수상관계수표명SPARC단백표체여폐포상피세포조망정정상관.결론 재소서폐섬유화진정중존재SPARC과표체,과표체적SPARC단백대폐섬유화가능기촉진작용.억제Rho/Rho격매신호통로가감소SPARC단백표체,재연완폐섬유화진정중가능기중요작용.
Objective To observe the changes of SPARC expression and intervention effect of fasudil in experimental mice induced by bleomycin,so as to discuss the action mechanism of SPARC in pulmonary fibrosis.Methods According to the random number table,all 108 mice were randomly divided into control group (n =36),model group (n =36) and fasudil group (n =36).The three groups were injected with equal 0.9 ％ sodium chloride injection and BLM (3.5 mg/kg) by disposable syringe firstly.The FAS group was injected with intraperitoneal FAS 15 mg/kg injection after BLM injection.Eight mice of each group were killed respectively at the 3rd,7th,14th,28th day and the lung tissues were made into pathological sections.Histological changes were observed by HE staining and the apoptosis level of lung tissue of BLM group and Cont group were tested by TUNEL,while western blot were used to detect the expression of SPARC and Rock-1 of the three groups.Moreover,the correlation coefficient between SPARC and apoptosis index was calculated.Results One hundred and eight mice were involved in the result analysis and eight mice of the model group died.① HE staining:the BLM lung tissue gradually changed from pulmonary alveolitis into fibrosis was observed.②The TUNEL:compared to those in the Cont group,the apoptosis index of lung cells in the BLM group was increasing progressively.③The result of Western blot presented that the expression of SPARC and Rock-1 of BLM increased significantly.Compared to the BLM group,the level of SPARC and Rock-1 expression were decreased gradually in the FAS group.The correlation coefficient between SPARC and AI in the BLM group suggests that there exists positive correlation between AI and the expression of SPARC in the model group.Conclusions There exists abnormal expression of SPARC in the progression of mice pulmonary fibrosis.The abnormal expression of SPARC may play a driving role in the pulmonary fibrosis.The dynamic expression of SPARC may have a negative effect on the cell cycle of alveolar epithelial cells.To suspend the progression of PF,it might be very important to restrain the Rho/Rho kinase signal pathway.