棒柄花叶提取物对肝纤维化大鼠的防治作用及其机制

Preventive effect of water extract of Cleidion brevipetiolatum leaves on hepatic fibrosis in rats and its mechanism

目的:探讨棒柄花叶水提物对四氯化碳(CCl4)诱导的实验性肝纤维化大鼠防治作用及其机制。方法:(1)SPF级SD雄性大鼠60只随机分为6组:正常对照组、模型对照组、秋水仙碱组和棒柄花叶水提物高、中、低剂量组(;2)除正常对照组外,各组灌胃给予50%CCl4花生油溶液(1 m L/kg),每周2次,连续6周;造模同时,秋水仙碱组灌胃0.2 mg/kg的秋水仙碱溶液,棒柄花叶水提物高、中、低剂量组分别灌胃4.8 g/kg、2.4 g/kg、1.2 g/kg的棒柄花叶水提物,模型对照组灌胃等体积蒸馏水,1次/d(;3)HE和Masson染色观察肝组织的损伤和纤维化程度;(4)全自动生化分析仪检测血清中的谷丙转氨酶(ALT)、谷草转氨酶(AST);(5)酶联免疫吸附测定法(ELISA)检测血清中三型胶原(COL-Ⅲ)、四型胶原(COL-Ⅳ)、肿瘤坏死因子(TNF-α)、白细胞介素-1(IL-1)的水平;(6)RT-PCR法检测大鼠肝组织PDGF、FAK、Akt mRNA的表达。结果:(1)与模型对照组相比,棒柄花叶水提物给药组对纤维化大鼠体质量、胸腺指数、肝脏指数有改善的趋势,明显降低ALT、AST、COL-Ⅲ、COL-Ⅳ、TNF-α、IL-1的含量和显著降低肝组织PDGF、FAK、Akt m RNA的表达水平(P<0.05或P<0.01)(;2)肝组织病理学显示棒柄花叶水提物给药组明显改善CCl4所致大鼠的肝纤维化损伤。结论:棒柄花叶水提物对CCl4诱导的大鼠肝纤维化具有明显的保护作用,能改善模型大鼠的肝功能及病理损伤,其作用机制可能与抑制肝脏胶原纤维生成、减轻炎症刺激、抑制PDGF-FAK/Akt信号通路有关。

Objective:To investigate the effect of water extract from the leaves of Cleidion brevipetiolatum on the prevention and treatment of carbon tetrachloride(CCl4)-induced experimental hepatic fibrosis in rats and its mechanism.Methods:(1)Sixty male SPF SD rats were randomly divided into six groups:normal control group,model control group,colchicine group and Cleidion brevipetiolatum leaf water extract low,medium and high dose groups.(2)Except for the normal control group,each group was given 50%CCl4arachis oil solution(1 mL/kg)by gavage twice a week for 6 weeks.At the same time,the colchicine group was gavaged with 0.2 mg/kg of colchicine solution,the high,medium and low dose groups were gavaged with 4.8 g/kg,2.4 g/kg and 1.2 g/kg of water extract from the leaves of Cleidion brevipetiolatum,respectively,and the model control group was gavaged with an equal volume of distilled water,all once per day.(3)HE and Masson staining were performed to observe the degree of liver tissue damage and fibrosis.(4)A fully automatic biochemical analyzer was used to detect the alanine transaminase(ALT)and aspartate aminotransferase(AST)in serum.(5)Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of collagen typeⅢ(COL-Ⅲ),collagen typeⅣ(COL-Ⅳ),tumor necrosis factor(TNF-α)and interleukin-1(IL-1)in serum.(6)RT-PCR was performed to detect mRNA expression of PDGF,FAK and Akt in rat liver tissues.Results:(1)Compared with the model control group,fibrotic rats in the Cleidion brevipetiolatum leaf water extract groups showed improvements in body mass,thymus indexes and liver indexes and had significantly reduced levels of ALT,AST,COL-Ⅲ,COL-Ⅳ,TNF-α,IL-1 and expression levels of PDGF,FAK and Akt mRNA in liver tissues(P<0.05 or P<0.01).(2)Liver histopathology showed that the water extract of Cleidion brevipetiolatum leaf significantly improved the CCl4-induced hepatic fibrosis injury in rats.Conclusion:Water extract of Cleidion brevipetiolatum leaves has significant protective effects on CCl4-induced hepatic fibrosis in rats,improving liver function and pathological injury in model rats.Its mechanism of action may be related to the inhibition of hepatic collagen fibrogenesis,reduction of inflammatory stimulation,and inhibition of PDGF-FAK/Akt signaling pathway.