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目的观察芍药苷对日本血吸虫感染小鼠肝脏结缔组织生长因子(CTGF)、血小板衍化生长因子(PDGF)蛋白表达水平和血清中肿瘤坏死因子α(TNF-α)水平的影响,探讨芍药苷预防血吸虫病肝虫卵肉芽肿及纤维化的作用机制。方法小鼠随机分为A～E5组,A组不感染血吸虫尾蚴,为正常对照组;B、C、D、E4组小鼠感染尾蚴后6周以吡喹酮灌胃杀虫,剂量为400mg/(kg·d),连续2d。B、C、D3组杀虫后给予芍药苷,剂量分别为30、60、120mg/(kg·d)),E组不给芍药苷,为感染对照组。5组小鼠均在芍药苷给药8周后处死,取血及肝脏。HE及Masson胶原纤维染色观察肝虫卵肉芽肿面积及纤维化程度,采用免疫组化技术检测CTGF和PDGF蛋白表达情况,采用ELISA法检测小鼠血清中TNF-α水平。结果芍药苷高剂量的D组与感染对照E组相比,肝组织中CTGF和PDGF蛋白表达明显减少,血清中TNF-α水平也明显下降(P均<0.05);而中、低剂量B组和C组的CTGF、PDGF和TNF-α水平亦较E组减少,但差异无统计学意义(P均>0.05)。结论芍药苷抗血吸虫病肝纤维化作用呈剂量依赖性,在吡喹酮杀虫处理基础上联合应用高剂量芍药苷,可通过减少肝组织CTGF和PDGF蛋白表达,降低血清中TNF-α水平,减轻肝纤维化程度。
Objective To observe the effect of paeoniflorin on the expression of hepatic connective tissue growth factor (CTGF), platelet-derived growth factor (PDGF) and serum tumor necrosis factor α (TNF-α) in mice infected with Schistosoma japonicum, Egg granuloma and the mechanism of fibrosis. Methods Mice were randomly divided into groups A to E5. Group A was not infected with cercariae of Schistosoma japonicum and was normal control group. The mice in groups B, C, D and E4 were injected intraperitoneally with praziquantel / (kg · d), continuous 2d. Paeoniflorin was given to mice in groups B, C and D3 after insecticide treatment, the doses were 30, 60, 120 mg / (kg · d), respectively). Five groups of mice were sacrificed 8 weeks after paeoniflorin administration, blood and liver were taken. Hematoxylin and eosin (Masson) collagen fibers were used to observe the granuloma area and the degree of fibrosis. The expression of CTGF and PDGF protein was detected by immunohistochemistry. The level of TNF-α in serum was detected by ELISA. Results Compared with the control group E, the expression of CTGF and PDGF in the group of high dose of paeoniflorin was significantly decreased and the level of TNF-α in the serum was significantly decreased (P <0.05), while the dose of medium and low dose of group B The levels of CTGF, PDGF and TNF-α in group C were also lower than those in group E, but the difference was not statistically significant (all P> 0.05). Conclusion Paeoniflorin has a dose-dependent effect on hepatic fibrosis induced by schistosomiasis. Combined with high dose of paeoniflorin on the basis of praziquantel insecticide treatment, paeoniflorin can reduce the expression of CTGF and PDGF in serum and decrease the level of TNF-α in serum, Reduce the degree of liver fibrosis.