CAJ | 학술논문

目的研究肥大细胞(MC)对实验性肝炎大鼠的影响及其作用机制.方法取Wistar大鼠30只,随机分为2组:正常对照(NC)组、慢性肝炎(CH)组.CH组采用复合因素造模(用40％四氯化碳油溶液皮下注射,同时辅以低蛋白、低胆碱、高脂肪、高醇饮食).4周末处死动物,取血分别检测血浆类胰蛋白酶(TS)、组织胺(HA)水平,同时观察肝脏组织学变化及MC形态改变.应用免疫组织化学方法观察肝脏c-kit和干细胞因子(SCF)的表达.用逆转录-多聚酶链反应(RT-PCR)方法观察肝组织c-kit mRNA和SCF mRNA的表达水平.组间比较采用SPSS统计学软件进行t检验,相关分析采用直线相关回归分析,P＜0.05为差异具有统计学意义.结果 (1)与正常对照组比较,CH组血和肝组织HA水平[(78.09±38.55)ng/ml对比(145.14±52.54)ng/ml和每100mg肝组织中(51.58±20.45) ng对比(106.59±43.15)ng,t值为2.457和2.825,P＜0.05],以及TS均有明显升高[(0.476±0.143)mg/ml对比(0.753±0.210) mg/ml,t值为4.165,P＜ 0.05].(2)光镜下,CH组有脂肪变性和纤维化形成；甲苯胺蓝染色可见CH组肝脏血管周围及纤维间隔内大量正在脱颗粒和已经脱颗粒的充满紫色颗粒的MC.CH组MC数目较正常对照组明显升高[(2.167±0.924)个/hp对比(10.92±1.575)个/hp,t=7.633,P＜0.05].(3)免疫组织化学染色结果显示CH组高表达c-kit和SCF(相对表达量2.783±0.577对比12.86±3.126和3.383±1.583对比15.58±6.431,t值分别为9.511和9.625,P＜0.05),而且二者的表达均与HA水平呈明显正相关(r值分别为0.478、0.556,P＜0.05).RT-PCR结果显示CH组c-kit mRNA和SCFmRNA表达水平明显增高.结论 CH时MC脱颗粒明显增加,HA释放增加,可能与膜受体c-kit及其配体SCF的表达增加有关.
목적 연구비대세포(MC)대실험성간염대서적영향급기작용궤제.방법 취Wistar대서30지,수궤분위2조:정상대조(NC)조、만성간염(CH)조.CH조채용복합인소조모(용40％사록화탄유용액피하주사,동시보이저단백、저담감、고지방、고순음식).4주말처사동물,취혈분별검측혈장류이단백매(TS)、조직알(HA)수평,동시관찰간장조직학변화급MC형태개변.응용면역조직화학방법관찰간장c-kit화간세포인자(SCF)적표체.용역전록-다취매련반응(RT-PCR)방법관찰간조직c-kit mRNA화SCF mRNA적표체수평.조간비교채용SPSS통계학연건진행t검험,상관분석채용직선상관회귀분석,P＜0.05위차이구유통계학의의.결과 (1)여정상대조조비교,CH조혈화간조직HA수평[(78.09±38.55)ng/ml대비(145.14±52.54)ng/ml화매100mg간조직중(51.58±20.45) ng대비(106.59±43.15)ng,t치위2.457화2.825,P＜0.05],이급TS균유명현승고[(0.476±0.143)mg/ml대비(0.753±0.210) mg/ml,t치위4.165,P＜ 0.05].(2)광경하,CH조유지방변성화섬유화형성；갑분알람염색가견CH조간장혈관주위급섬유간격내대량정재탈과립화이경탈과립적충만자색과립적MC.CH조MC수목교정상대조조명현승고[(2.167±0.924)개/hp대비(10.92±1.575)개/hp,t=7.633,P＜0.05].(3)면역조직화학염색결과현시CH조고표체c-kit화SCF(상대표체량2.783±0.577대비12.86±3.126화3.383±1.583대비15.58±6.431,t치분별위9.511화9.625,P＜0.05),이차이자적표체균여HA수평정명현정상관(r치분별위0.478、0.556,P＜0.05).RT-PCR결과현시CH조c-kit mRNA화SCFmRNA표체수평명현증고.결론 CH시MC탈과립명현증가,HA석방증가,가능여막수체c-kit급기배체SCF적표체증가유관.
Objective To study the potential role of mast cells and the related molecular mechanism in chronic hepatitis (CH) using a rat model system.Methods Thirty Wistar mts (15 males,15 females; weight range:230-290 g) were randomly divided into the normal contrast (NC) group and experimental CH group.The CH group received subcutaneous injection of CC14 and a diet high in cholesterol and alcohol content and low in protein and choline content.Throughout the 4-week modeling period,aseptic blood samples were taken to test plasma tryptase (TS) and hyaluronic acid (HA) levels.The rats were euthanized to assess the changes in liver mast cells by histology and morphology analyses and the changes in liver expression of c-kit and stem cell factor (SCF) proteins by immunohistochemistry and mRNAs by RTPCR.Results Compared to the NC group,the CH group had higher plasma and liver concentration of HA (78.09 ± 38.55 vs.145.14 ± 52.54 ng/ml,51.58 ± 20.45 vs.106.59 ± 43.15 ng/l00 mg; t =2.457 and 2.825respectively,both P ＜ 0.05) and TS (0.416 ± 0.143 vs 0.753 ± 0.210 mg/ml; t =4.165,P ＜ 0.05).The CH group also showed fatty degeneration and fibrosis with many degranulating and degranulated mast cells filled with purple granula located around the liver blood vessels and in fiber-intervals.The CH livers also showed a significantly higher number of mast cells (2.167 ± 0.924 vs.NC:10.92 ± 1.575; t =7.633,P ＜0.05) and stronger intensity of c-kit staining (2.783 ± 0.577 vs.12.86 ± 3.126; t =9.511,P ＜ 0.05) and SCF staining (3.383 ± 1.583 vs.15.58 ± 6.431; t =9.625,P ＜ 0.05).The expressions of c-kit and SCF were positively correlated with HA level (r =0.478 and 0.556 respectively,both P ＜ 0.05).The c-kit and SCF mRNA expression levels were also significantly higher in the CH liver tissues.Conclusion Mast cell degranulation and histamine release is significantly increased under conditions of chronic hepatitis,and the related mechanism may involve up-regulation of the membrane receptor c-kit and its ligand SCE.