Effect of Prunellae Spica Sulfated Polysaccharide on Carbon Tetrachaloride (CCl)-induced Hepatic Fibrosis and Activation of Hepatic Stellate Cell Induced by TGF-

FU Yue-yue; ZHU Lan-ping; ZHANG Guo-liang; ZHANG Shuang-xia

doi:10.13422/j.cnki.syfjx.20181056

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Effect of Prunellae Spica Sulfated Polysaccharide on Carbon Tetrachaloride (CCl)-induced Hepatic Fibrosis and Activation of Hepatic Stellate Cell Induced by TGF-

更新时间：2024-01-04

- Effect of Prunellae Spica Sulfated Polysaccharide on Carbon Tetrachaloride (CCl)-induced Hepatic Fibrosis and Activation of Hepatic Stellate Cell Induced by TGF-
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 14, Pages: 147-152(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20181056
  CLC： R285.5
- Published：2018
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FU Yue-yue, ZHU Lan-ping, ZHANG Guo-liang, et al. Effect of Prunellae Spica Sulfated Polysaccharide on Carbon Tetrachaloride (CCl)-induced Hepatic Fibrosis and Activation of Hepatic Stellate Cell Induced by TGF-[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 147-152.
DOI：

FU Yue-yue, ZHU Lan-ping, ZHANG Guo-liang, et al. Effect of Prunellae Spica Sulfated Polysaccharide on Carbon Tetrachaloride (CCl)-induced Hepatic Fibrosis and Activation of Hepatic Stellate Cell Induced by TGF-[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 147-152. DOI： 10.13422/j.cnki.syfjx.20181056.

摘要

目的：观察夏枯草硫酸多糖(PSSP)对四氯化碳(CCl4)所致的大鼠肝纤维化及转化生长因子-β1(TGF-β1)诱导活化大鼠肝星状细胞-T6(HSC-T6)的影响，从而初步探讨PSSP抗纤维化的机制。方法：采用CCl4-橄榄油溶液腹腔注射诱导大鼠肝纤维化造模，随机分为5组，分别为正常组，模型组，秋水仙碱组(0.25 mg·kg-1)，PSSP高剂量组(300 mg·kg-1)和PSSP低剂量组(100 mg·kg-1)。分别测定各组大鼠血清丙氨酸氨基转移酶(ALT)，天门冬氨酸氨基转移酶(AST)，层粘连蛋白(LN)，透明质酸(HA)，Ⅲ型前胶原(PCⅢ)和Ⅳ胶原(C-Ⅳ)的含量；采用苏木素-伊红(HE)及天狼猩红染色观察大鼠肝组织病理学改变；体外培养HSC-T6细胞；采用细胞增殖与活性法(CCK-8)测定PSSP对TGF-β1诱导的HSC-T6细胞增殖的影响；采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测HSC-T6细胞中Ⅰ型胶原(Col-I)，α-平滑肌肌动蛋白(α-SMA) mRNA和蛋白的表达。结果：与模型组比较，PSSP可明显降低大鼠ALT，AST，HA，LN，PC-Ⅲ和C-Ⅳ的含量，并减轻大鼠肝纤维化程度(P<0.01)；PSSP可抑制TGF-β1诱导的HSC-T6细胞的增殖，减少HSC-T6细胞Col-I，α-SMA mRNA和蛋白的相对表达量(P<0.01)。结论： PSSP具有很好的抗纤维化作用，其机制可能与抑制肝星状细胞活化及抑制胶原合成与沉积，减少细胞外基质(ECM)的生成并促进其降解有关。

Abstract

Objective: To investigate the effect of Prunellae Spica sulfated polysaccharide (PSSP) on carbon tetrachaloride (CCl4)-induced hepatic fibrosis and activation of HSC-T6 induced by transforming growth factor(TGF)-β1. Method: The liver fibrosis model was established through intraperitoneal injection with 40% CCl4.The rats were randomly divided into five groups:normal control group

model group

positive control group (colchicines

0.25 mg·kg-1)

and high and low-dose PSSP groups. Serum levels of alanine aminotransferase(ALT)

aspartate aminotransferase(AST)

hyaluronic acid(HA)

laminin(LN)

procollagen-Ⅲ-peptide(PC-Ⅲ) and type Ⅳ collagen(C-Ⅳ) were examined in different groups. Moreover

htoxylin eosin(HE) staining and sinus red staining were used to examine the degree of hepatic fibrosis. HSC-T6 was cultured

and the proliferation of HSC-T6 was determined by cell counting kit-8(CCK-8) assay. The expressions of collagen type I (Col-I) and α-smooth muscle actin (α-SMA) mRNA were detected by Real-time PCR. The mRNA and protein expressions of Col-I and α-SMA were measured by Western blot. Result: Compared with the model group

the levels of ALT

AST

PC-Ⅲ and C-Ⅳ were significantly decreased in the PSSP groups. HE staining and sinus red staining showed that PSSP could significantly reduce the degree of hepatic fibrosis. PSSP had an inhibitory effect on the proliferation of TGF-β1-induced HSC-T6 cells(P<0.01). Compared with TGF-β1 group

mRNA and protein expressions of α-SMA and Col-I were significantly decreased by PSSP in TGF-β1-induced HSC-T6 cells(P<0.01). Conclusion: PSSP has a protective effect against CCl4-induced liver fibrosis possibly by reducing secretion of collagen and promoting extracellular matrix degradation.

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