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目的:观察两头尖提取物对猪血清免疫诱导大鼠肝纤维化模型的干预作用。方法:SD大鼠68只随机分为5组,即空白组、模型组、两头尖提取物组(以下简称提取组)、两头尖原药组(以下简称原药组)、扶正化瘀组(以下简称扶正组)。采用猪血清腹腔注射法制备大鼠肝纤维化模型,0.5 mL/只,每周2次,连续注射15周。第15周模型成功后,开始灌胃给药,扶正组给予扶正化瘀胶囊(0.525 g.kg-1),原药组给予两头尖煎液(0.7 g.kg-1),提取物组给予两头尖提取物(0.071 g.kg-1),模型组与空白组灌服等量生理盐水,日1次,疗程8周。大鼠于第23周末处死取材,分别测定血清丙氨酸氨基转移酶(ALT),天冬氨酸氨基转移酶(AST),白蛋白(Alb),透明质酸(HA),层黏蛋白(LN),Ⅲ型前胶原(PCⅢ),Ⅳ型胶原(CⅣ)含量与肝组织中羟脯氨酸(Hyp)含量。HE染色与Masson法检测肝组织病理;免疫组化法(SABC法)测定肝组织中Ⅰ型胶原(col-Ⅰ)、Ⅲ型胶原(col-Ⅲ)表达。结果:提取组、原药组血清ALT,AST,Alb,HA,PCⅢ,CIV水平,肝组织Hyp含量分别为(79.77±14.68),(75.20±11.21),(168.22±19.46),(173.72±18.52)U.L-1,(27.40±1.78),(26.95±2.14)g.L-1,(52.36±5.12),(43.29±3.56),(51.63±6.41),(52.18±4.79),(14.92±2.26),(13.93±1.88)μg.L-1,(1.04±0.12),(1.15±0.06)mg.g-1。与模型组相比,提取组、原药组可显著降低血清ALT,AST,HA,PCⅢ,CIV水平与肝组织Hyp含量(P<0.05),升高血清Alb水平(P<0.05)。与模型组相比,提取组、原药组炎症活动度、纤维化程度显著减轻。Ⅰ型胶原、Ⅲ型胶原表达面积和强度明显为弱,纤维间隔染色淡,无典型假小叶形成。显色指数结果显示,提取组、扶正组肝组织中col-Ⅰ,col-Ⅲ表达均明显减少。结论:采用猪血清免疫法可成功制备肝纤维化大鼠模型,两头尖原药及其提取物具有良好的抗肝纤维化作用,且提取物抗肝纤维化作用基本等同于两头尖原药,推测其作用机制为抑制ECM合成并促进ECM降解有关。
OBJECTIVE: To observe the intervention effect of two sharp extracts on rat model of hepatic fibrosis induced by immune of pigs. Methods: Sixty-eight SD rats were randomly divided into five groups: blank group, model group, two-point extract group (hereinafter referred to as extraction group), two-point original drug group (hereinafter referred to as the original drug group), Fuzhenghuayu group Hereinafter referred to Fuzheng group). Rat model of hepatic fibrosis was established by intraperitoneal injection of porcine serum, 0.5 mL per animal twice a week for 15 weeks. At the 15th week after the model was successfully administered, intragastric administration was started, and Fuzheng Huayu Capsule (0.525 g.kg-1) was given to Fuzheng group. The two groups of Jianjianhuayu capsules (0.7 g.kg-1) Two tip extracts (0.071 g.kg-1), the model group and the blank group were given the same amount of saline, once a day for 8 weeks. The rats were sacrificed on the 23rd weekend and the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), hyaluronic acid (HA), laminin LN), type Ⅲ procollagen (Ⅲ), type Ⅳ collagen (C Ⅳ) and content of hydroxyproline (Hyp) in liver tissue. The liver histopathology was examined by HE staining and Masson’s method. The expression of col-Ⅰ and col-Ⅲ was detected by immunohistochemistry (SABC). Results: The levels of serum ALT, AST, Alb, HA, PCⅢ and CIV in the extract group and the original drug group were (79.77 ± 14.68), (75.20 ± 11.21), (168.22 ± 19.46) and (173.72 ± 18.52 ) Were significantly higher than those in the control group (P <0.05). UL-1, (27.40 ± 1.78), (26.95 ± 2.14) gL- (13.93 ± 1.88) μg.L-1, (1.04 ± 0.12) and (1.15 ± 0.06) mg.g-1, respectively. Compared with the model group, the levels of serum ALT, AST, HA, PCⅢ, CIV and the content of Hyp in the liver tissue were significantly decreased (P <0.05), and the levels of serum Alb were increased significantly (P <0.05). Compared with the model group, the degree of inflammation and fibrosis in the extraction group and the original drug group were significantly reduced. Type Ⅰ collagen, type Ⅲ collagen expression area and intensity was significantly weaker, fibrous staining pale, no typical pseudolobule formation. The results of color rendering index showed that the expression of col-Ⅰ and col-Ⅲ in the liver tissue of the extraction group and Fuzheng group were significantly decreased. Conclusion: The rat model of hepatic fibrosis can be successfully prepared by swine serum immunization. The two traditional Chinese medicines and their extracts have a good anti-hepatic fibrosis effect, and the anti-hepatic fibrosis effect of the extract is basically equivalent to the two- It is speculated that its mechanism of action is to inhibit ECM synthesis and promote ECM degradation.