摘要
目的:探讨中药复方补肾柔肝方对二甲基亚硝胺(dimethylnitrosamine,DMN)诱导大鼠肝纤维化后肝脏组织结缔组织生长因子(connective tissue growth factor,CTGF)mRNA的表达影响,以了解其对肝纤维化的治疗作用和分子机制.方法:♂Wistar大鼠40只,随机分为正常对照组(n=10)、模型组(n=15)及治疗组(n=15).模型组和治疗组以10mg/kg的剂量腹腔注射DMN,每天1次,每周连续3d,共4wk.模型组在造模结束后给予生理盐水ig,而治疗组在造模结束后给补肾柔肝方ig进行治疗干预,用药4wk.第8周末处死全部大鼠,用放射免疫试剂盒方法检测血清胶原成分HA、LN及Ⅳ-C的含量;用HE和天狼猩红染色法观察肝组织的炎症及纤维增生情况.并采用RT-PCR半定量方法,探讨大鼠肝组织CTGFmRNA的表达水平.结果:治疗组大鼠一般状态明显好于模型组正常组大鼠无死亡,模型组死亡率为40%,治疗组死亡率20%.模型组血清胶原成分HA、LN及Ⅳ-C的含量较正常组显著增高,治疗组较模型组显著下降(HA:319.75±63.23pg/Lvs434.44±98.81pg/L;LN:44.83±4.09pg/Lvs70.67±6.32pg/L;Ⅳ-C:52.79±5.71pg/Lvs79.39±10.52pg/L,均P<0.01).DMN可以成功诱导大鼠肝纤维化,模型组肝组织CTGFmRNA表达明显增强,中药复方治疗组与模型组相比明显减弱(CTGF/β-actin:0.76±0.10vs1.08±0.17,P<0.01),而正常对照组表达较少.结论:CTGF mRNA的表达可能与肝纤维化的发生密切相关,中药复方补肾柔肝方具有较好的抗肝纤维化作用,可以显著抑制大鼠肝组织CTGF mRNA的表达.
AIM: To investigate effects of kidney-tonifying liver-emoliating formula (KTLEF) on expression of connective tissue growth factor (CTGF) mRNA in dimethylnitrosamine-induced hepatic fibrosis rats and thereby to elucidate its therapeutic effects and its underlying molecular mechanism. METHODS: Forty male Wistar rats were ran- domly assigned to normal control group (n = 10), model group (n = 15) and KTLEF-treated group (n = 15). Except the normal control group, all the rats received intraperitoneal DMN injec- tion once a day for 3 successive days for 4 wk. Then only the model group was given KTLEF for anther 4 wk. Rats were all executed at week 8. The serum liver fibrosis markers, such as HA, LN and IV-C, were measured using ELISA and RIA. The Hepatic inflammatory necrosis and col- lagen deposition were determined by HE stain- ing and Sirius red staining, and CTGF mRNA expression was detected using RT-PCR. RESULTS: The rat model of liver fibrosis in- duced by DMN was successfully constructed. Serum HA, LN and IV-C levels were significantly declined in BSRGF-treated group compared with those in the model-group (HA: 319.75 ± 63.23 pg/ L vs 434.44 ± 98.81 pg/L; LN: 44.83 ± 4.09 pg/L vs 70.67±6.32 pg/L; IV-C: 52.79 ± 5.71 pg/L vs 79.39 ± 10.52 pg/L, all P 〈 0.01). The expression level of CTGF mRNA was lower in the KTLEF-treated group than that in the fibrosis model group (CTGF/β-actin: 0.76 ± 0.10 vs 1.08 ± 0.17, P 〈 0.01), and the least in the normal control group. CONCLUSION: The expression of CTGF mRNA is increased in the hepatic fibrosis rats, and is supposed to be one possible mechanism of hepatic fibrosis. KTLEF can significantly inhibit CTGF mRNA expression and then effectively counteract hepatic fibrosis.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第20期2224-2228,共5页
World Chinese Journal of Digestology
关键词
补肾柔肝方
肝纤维化
二甲基亚硝胺
结缔组织生长因子
动物
实验
Kidney-tonifying liver-emoliating formula
Hepatic fibrosis
Dimethylnitrosamine
Connective tissue growth factor
Animals
Laboratory