“肝心宁”防治肝纤维化的作用及机理研究

【作者】 李生财；

【导师】 谢春光； 王再谟；

【作者基本信息】 成都中医药大学 ， 中医内科学， 2008， 博士

【摘要】 目的:观察“肝心宁”对实验大鼠肝纤维化的防治作用,研究“肝心宁”对实验大鼠肝纤维化的防治作用的机理,为“肝心宁”在临床的广泛应用提供理论依据。方法:57只sD大鼠随机分为空白组、模型组、阳性对照组、中药低剂量组、中药高剂量组5组。采用改良复合因素法复制大鼠肝纤维化模型。空白组与模型组给予生理盐水10ml/kg灌胃,阳性对照组给予秋水仙碱0.125mg/kg治疗,中药高、低剂量组分别给予肝心宁26g/kg、6.5g/kg治疗。6周后,HE染色后比较各组大鼠肝脏病理组织改变;采用全自动生化分析仪,对各组大鼠血清ALT、AST、TBA进行检测;采用放免分析法检测大鼠血清HA、PⅢNP、LN的含量;用免疫组织化学法研究各组大鼠肝脏组织中caspase-3、bcl-2表达的部位及强度;并测定肝组织中PDGF-BB、bFGF的表达情况。结果:在肝脏病理积分上各治疗组与模型组比较均有显著性差异(P＜0.05或P＜0.01),且在炎症积分上中药低剂量组效果最为明显,与阳性对照组比较也有显著性差异(P＜0.05);在血清酶学指标上,与空白组比较,其余各组大鼠血清中ALT、AST的活性与TBA的含量均明显升高,有显著性差异(P＜0.01或P＜0.05)。与模型组比较,各治疗组血清中ALT、AST的活性与TBA的含量均明显减少有显著性差异(P＜0.01或P＜0.05),各治疗组间无显著性差异(P＞0.05);在血清ECM指标上,与空白组比较,其余各组大鼠血清中LN、HA、PⅢNP的含量均明显升高,有显著性差异(P＞0.01或P＜0.05),与模型组比较,各治疗组血清中LN、HA、PⅢNP的含量均明显减少,有显著性差异(P＜0.01或P＜0.05)。治疗药物之间,除血清LN中药低剂量组显著低于阳性对照组(P＜0.05)外,其余各治疗组之间比较无显著性差异(P＞0.05);在抑制bFGF表达上,与模型组比较,各治疗组肝组织中bFGF均明显减少,有显著性差异(P＜0.01),各治疗组间无显著性差异(P＞0.05);在抑制PDGF-BB表达上,各治疗组与模型组比较均有显著性差异(P＜0.01),各治疗组间无显著性差异(P＞0.05);在调节caspase-3方面,与模型组比较,各治疗组肝组织中caspase-3均明显减少,有显著性差异(P＜0.01或P＜0.05)。,治疗药物之间,中药低剂量组显著低于阳性对照组(P＜0.05),肝心宁低剂量组、肝心宁高剂量之间比较无显著性差异(P＞0.05);在调节bcl-2方面,与空白组比较,其余各组各组大鼠肝组织中bcl-2均明显升高,有显著性差异(P＜0.01),与模型组比较,各治疗组肝组织中bcl-2未见明显减少(P＞0.05)。结论:“肝心宁”能有效预防实验大鼠肝纤维化形成,改善肝纤维化大鼠肝组织损伤程度,可以有效降低肝纤维化模型大鼠血清ALT、AST、TBA等酶学指标和LN、HA、PⅢNP等肝纤维化指标的含量。其机制可能与“肝心宁”能够抑制PDGF-BB、bFGF介导的HSC增殖有关;此外通过调整caspase-3,bcl-2减轻肝细胞凋亡也可能“肝心宁”防治肝纤维化机制之一。中药防治肝纤维化往往是通过多途径、多层次、多靶点来实现的,所以以上结论的可靠性还需要在以后的科研临床工作中进一步验证。更多还原

【Abstract】 Objective:To observe the preventive and therapeutic effect of "Ganxinning" capsule on experimental HF rats and the possible therapeutic mechanism,to provide the theoretic evidence for the clinic apply of"Ganxinning".Methods:57 SD rats were divided into 5 groups randomly,the normal group,the model group,the positive control group,the high dose group,and the low dose group.HF rat model was duplicated with Improved Composite Factor Method.Rats of the normal group and the model group were administered normal sodium with dose 10ml/kg,positive control group were administered Colchicine with the dose 0.125mg/kg.The high dose group and the low dose group were administered "Ganxinning" with the dose 26g/kg、6.5g/kg respectively.The liver pathology change of each group by HE coloration were compared and determination of liver tissue PDGF-BB expression 6 weeks later.The ALT,AST,and TBA in blood serum of experimental rats were detected by automatic biochemical analyzer.The HA,PⅢNP and LN were detdected by radio-immunoassay.The expression of caspase-3,bcl-2,PDGF-BB and bFGF were detected by immunohistochemical analyzer,Results:Compared with the model group,each treatment group has significant difference in the liver pathology integral(P＜0.05 or P＜0.01),and the low dose group is the most significant difference in the inflammation integral.Compared with the Colchicin group,each treatment group has significant difference too(P＜0.05).Compared with the control group,the ALT,AST and TBA of each group increased significantly(P＜0.05 or P＜0.01).And compared with the model group,these indexes of each treatment group decreased significantly(P＜0.01 or P＜0.05).these indexes between each treatment group showed no significant difference(P＞0.05).Compared with the control group,the LN,HA and PⅢNP of each group increased significantly(P＜0.05 or P＜0.01).And compared with the model group, these indexes of each treatment group decreased significantly(P＜0.01 or P＜0.05).The LN of lower dose TCM group was lower significantly than the Clochicin group(P＜0.05).these indexes between the other treatment groups showed no significant difference.Compared with the model group,the expression of bFGF of each treatment group decreased significant(P＜0.01).the expression of bFGF between each treatment group showed no significant difference(P＞0.05).Compared with the model group,the expression of PDGF-BB of each treatment group decreased significant(P＜0.01).the expression of PDGF-BB between each treatment group showed no significant difference(P＞0.05).Compared with the model group,the caspase-3 in olive of each treatment group decreased significant(P＜0.01 or P＜0.05). The caspase-3 of lower dose TCM group was lower significantly than the Clochicin group(P＜0.05).The caspase-3 of the higher Ganxinning dose and lower Ganxinning dose showed no significant difference(P＞0.05).Compared with the control group,the bcl-2 of each treatment group increased significant(P＜0.01).Compared with the model group,thebcl-2 between each treatment group showed no significant difference(P＞0.05).Conclusion:"Ganxinning"capsule can prevent HF of the experimental rats effectively and decrease the enzymology index of ALT,AST,TBA and the HF index of LN,HA,PⅢNP.The possible mechanism is that Ganxinning can depress the HSC multiplication which were transfected by PDGF and bFGF. The other possible mechanism is that it can adjust the caspase-3、bcl-2 to prevent the apoptosis of liver cell.The TCM was used on HF mostly through multi-approach,multi-hiberarchy and multi-target.All of these experiment data need to be verified by clinical study.更多还原