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纸质出版日期:2018
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王乾宇, 王文佳, 奚锦, 等. 杜仲多糖对肝纤维化模型大鼠Ⅰ,Ⅲ型胶原蛋白,MMP-1,TIMP-1及TGF- mRNA表达的影响[J]. 中国实验方剂学杂志, 2018,24(23):153-158.
WANG Qian-yu, WANG Wen-jia, XI Jing, et al. Effect of Polysaccharides from Eucommiae Cortex on Expressions of Genes of Ⅰ, Ⅲ Collagen, MMP-1, TIMP-1 and TGF- from Hepatic Fibrosis in Rat Models[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 153-158.
王乾宇, 王文佳, 奚锦, 等. 杜仲多糖对肝纤维化模型大鼠Ⅰ,Ⅲ型胶原蛋白,MMP-1,TIMP-1及TGF- mRNA表达的影响[J]. 中国实验方剂学杂志, 2018,24(23):153-158. DOI: 10.13422/j.cnki.syfjx.20182334.
WANG Qian-yu, WANG Wen-jia, XI Jing, et al. Effect of Polysaccharides from Eucommiae Cortex on Expressions of Genes of Ⅰ, Ⅲ Collagen, MMP-1, TIMP-1 and TGF- from Hepatic Fibrosis in Rat Models[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(23): 153-158. DOI: 10.13422/j.cnki.syfjx.20182334.
目的:探讨杜仲多糖治疗肝纤维化(HF)作用,并探讨其相关的作用机制。方法:将60只健康SD大鼠随机分成2组,正常组(10只)和肝纤维化造模组(50只)。造模组采用40%四氯化碳(CCl4)腹腔注射制备HF动物模型,造模成功后将其随机分为5个组,分别为模型组,杜仲多糖高、中、低剂量组及秋水仙碱组每组10只。秋水仙碱组(1.0×10-4g·kg-1),杜仲多糖高、中、低剂量组(0.14,0.07,0.035 g·kg-1)分别连续灌胃给药8周后收集样本,测定法大鼠体质量及计算肝脏系数;采用酶联免疫吸附测定(ELISA)检测血清白细胞介素-6(IL-6),高迁移率族蛋白B1(high-mobility group box 1,HMGB1),脂多糖(LPS),肝组织基质金属蛋白酶-1(MMP-1)和金属蛋白酶组织抑制因子-1(TIMP-1)水平;逆转录聚合酶链式反应(RT-PCR)技术分析各组大鼠肝组织Ⅰ,Ⅲ型胶原蛋白,MMP-1,TIMP-1及转化生长因子-β1(TGF-β1)mRNA表达情况。结果:与正常组比较,CCl4能显著降低实验大鼠体质量(P<0.01),提高肝脏系数(P<0.01);杜仲多糖能显著增加HF模型的体质量(P<0.01),显著降低HF模型肝脏系数(P<0.01);ELISA检测表明,杜仲多糖能显著降低肝纤维化大鼠血清IL-6,HMGB1及LPS含量(P<0.01);RT-PCR检测结果显示,杜仲多糖及秋水仙碱能显著降低HF肝脏中Ⅰ,Ⅲ型胶原蛋白,TIMP-1及TGF-β1 mRNA含量(P<0.05),明显提高MMP-1含量(P<0.05,P<0.01),且呈量效关系。结论:杜仲多糖具有显著的抗肝纤维化作用,其作用机制可能与抑制降低HF肝脏中Ⅰ,Ⅲ型胶原蛋白,TIMP-1及TGF-β1 mRNA表达有关。
Objective:To investigate the therapeutic effect of polysaccharides from Eucommiae Cortex on liver fibrosis
in order to provide the experimental basis for the development of new anti-liver fibrosis drugs. Method:The 60 healthy SD rats were randomly divided into 2 groups
namely the normal control group (n=10) and the hepatic fibrosis(HF) group (n=50). The model of liver fibrosis was prepared through intraperitoneal injection with 40%CCl4 in the model group. The model was randomly divided into 5 groups:the model control group
the high
middle and low-dose polysaccharides from Eucommiae Cortex groups and the colchicine group
with 10 rats in each group. After successful modeling
colchicine group (1×10-4g·kg-1)
and high
medium and low-dose polysaccharides from Eucommiae Cortex groups (0.14
0.07
0.035 g·kg-1) were given drugs by gavage for 8 weeks
and then samples were collected to determine the mass of rats and calculate the liver coefficient. The levels of serum interleukin-6 (IL-6)
high-mobility group box 1 (HMGB1)
lipopolysaccharide (LPS) and the levels of matrix metalloproteinse-1(MMP-1) and tissue inhibitors of metalloproteinase-1(TIMP-1) in liver tissues were detected by enzyme-linked immunosorbent assay(ELISA). The expressions of Ⅰ
Ⅲ type collagen
MMP-1
TIMP-1 and transforming growth factor-β1(TGF-β1) in liver tissues of rats were analyzed by reverse transcription-polymerase chain reaction(RT-PCR) technique. Result:CCl4 can significantly reduce the body mass (P<0.01) of experimental rats and increase the liver coefficient (P<0.01). Polysaccharides from Eucommiae Cortex can significantly increase the body mass of the HF model (P<0.01)
and significantly lower the HF model liver coefficient (P<0.01). ELISA detection showed that polysaccharides from Eucommiae Cortex can significantly reduce IL-6
HMGB1 and LPS content (P<0.01) in the serum of liver fibroid rats. Polysaccharides from Eucommiae Cortex and colchicine could significantly reduce the mRNA content of Ⅰ
Ⅲ type collagen
TIMP-1 and TGF-β1 (P<0.05
P<0.01) in rat liver
and significantly increase the content of MMP-1 (P<0.05
P<0.01)
with a dose-effect relationship. Conclusion:Polysaccharides from Eucommiae Cortex had a significant anti-hepatic fibrosis effect. The mechanism may be related to the inhibition of the expressions of I
type Ⅲ collagen
TIMP-1 and TGF-β1 in HF liver.
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