摘要
目的:观察复方鳖甲软肝方药物血清对离体肝星形细胞(hepaticstellatecell,HSC)中Ⅰ,Ⅲ型胶原及组织金属蛋白酶抑制物(tissueinhibitorofmetalloproteinases,TIMPs)表达的影响,以探讨防治肝纤维化作用机制。方法:将成年健康雄性SD大鼠75只,随机分为正常对照组、模型组、高、中、低剂量药物组共5组,并制备各组血清。采用SD大鼠肝星形细胞分离、培养,药物血清制备,免疫组化检测实验各组肝星形细胞中Ⅰ,Ⅲ型胶原及TIMPs的表达经图像分析系统定量分析。结果:培养72h,高、中、低剂量药物组Ⅲ型胶原的表达犤Ⅲ型胶原反应物吸光度(A值)为178.36±5.82,182.27±20.32,220.38±19.32犦早于Ⅰ型胶原(I型胶原反应物A值为300.23±25.80,329.37±45.98,425.38±19.32),与模型组比较,差异有显著性意义(P<0.05)。高、中剂量药物组均可有效抑制Ⅰ,Ⅲ型胶原的表达,并呈药物量效关系;培养24h,实验各组均无TIMPs的表达,培养48h后TIMPs开始表达,培养72h时,高、中、低药物组与模型组比较,差异有显著性意义(P<0.05),可显著抑制TIMPs的表达。结论:复方鳖甲软肝方药物血清能有效抑制肝纤维化时肝星形细胞中Ⅰ,Ⅲ型胶原及TIMPs的表达,其抗肝纤维化作用机制可能与减少细胞外基质生成的同时,加速已生成的细胞外基质降解有关。
AIM:To observe the influence of fufang biejia rugan fang on the expression of typeⅠcollagen,type Ⅲcollagen and tissue inhibitor of metalloproteinases(TIMPs) of hepatic stellate cell(HSC)so as to explore the mechanism of antifibrosis of liver. METHODS:A total of 75 adult healthy male SD rats were randomly divided into five groups:normal control group,model group,high dosage drug group,medium dosage drug and low dosage drug serum group,with the serum prepared.HSCs in SD rats were separated and cultured,and meanwhile,the drug serum was prepared.The expression of typeⅠcollagen,type Ⅲcollagen and TIMPs of HSCs were measured by immunohistochemistry,and analyzed quantitively with image analysis system(IAS). RESULTS:Seventy two hours after cultivation,the expression of type Ⅲcollage in the high, medium and low dosage drug groups[type Ⅲcollagen reactant absorbency(A value) was 178.36±5.82,182.27±20.32,220.38±19.32] appeared earlier than those of typeⅠcollagen(type Ⅰcollagen reactant A value was 300.23±25.80,329.37±45.98,425.38±19.32),which indicated significant difference as compared with those of the model group(P< 0.05).Type Ⅰand Ⅲcollagen in the high and medium dosage drug groups could be effectively inhibited at a quantum effect manner;24 hours after cultivation,no TIMPs expression occurred in all group.After 48 hours,TIMPs expression appeared.After 72 hour cultivation,the comparison between high,medium,low dosage drugs and model group indicated significant difference(P< 0.05),and TIMPs expression was inhibited significantly. CONCLUSION:Fufang biejia ruangan fang can effectively inhibit the expression of typeⅠcollage,type Ⅲcollage and TIMPs in HSCs at hepatic fibrosis.The mechanism against hepatic fibrosis may be related to the reduction of synthesis of extracellular matrix(ECM) and the acceleration of degradation of ECM.
出处
《中国临床康复》
CSCD
2004年第27期5890-5893,i002,共5页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金重点资助项目(30130220)~~
