| Objective: Hepatic fibrosis is a chronic progressive disease characterized by increased extracellular matrix(ECM) synthesis and deposition in the interstitial space with parenchymal cell injury. The disequilibrium between collagen synthesis and degradation plays a pivotal role in fibrogenesis, although the mechanism of fibrogenesis has not been fully explained yet. Recent studies have indicated that collagenolytic potence was decreased in fibrotic liver, accompanied by elevated TIMP-1 as well as TIMP-2 level, and the level of TIMP-1 seems to be the most important. At the same time, TGF P 1 can stimulate the production of TIMP-1. Both TIMP-1 and TGF P 1 mainly derived from hepatic stellate cells(HSCs).It was reported that the active ingredient extracted from salvia miltiorrhiza, salvia miltiorrhiza and the compounds in which salvia miltiorrhiza serves as chief ingredient may prevent and treat rat hepatic fibrosis induced by CC14 through a mechanism of inhibiting collagen synthesis and HSCs proliferation, decreasing the deposition of ECM as well. The effects of salvia miltiorrhiza monomer IH764-3 on the degradation of collagen are still largely unknown. Its effectson expression of TIMP-1 and TGF 1 in HSCs and hepatic fibrosis induced by bile duct ligation(BDL) are poorly documented as well. The aims of this study were to probe roles of salvia miltiorrhiza monomer IH764-3 in anti-fibrogenesis. The machinism of it involving in collagen degradation was studied as well by experiments in vivo and in vitro.Methods: The experimental hepatic fibrosis was induced by common bile duct ligation. In brief, the rats were anesthetized with Ketamine in dose of 100mg .kg-1. In sham operation group, the common bile duct was dissected free without any other treatment. The common bile duct was double ligated and cut off between the ligation in the model group and IH764-3 group.Fourty male Spregue Desley rats were divided into three groups at random, namely control group (sham operation group) 10 rats, model group (operation group) 15 rats and IH764-3 group (operation+IH764-3) 15 rats. IH764-3 was diluted to 8g.L-1 by saline. The rats of treatment group received 40mg kg-1 of IH764-3 introperitoneally, while other rats were injected introperitoneally by equivalent saline. The study continued for 14 days. At the seventh day after operation, the body weight of rats was measured and the dose of IH764-3 and saline was adjusted correspondingly. At the end point of experiment, animals were sacrificed. The liver specimens were stored at -70 until hydroxyproline assayor fixed in 4% phosphate-buffered paraformaldehyde and embedded in paraffin. Tissue sections were stained with hematoxylin-eosin, Masson trichome. Other tissue sections were stained immunohistochemstrically to detect TIMP-1 and TGF 1 and positive area was messured.CFSC line, kindly provided by Prof. Greenwel, was derived from rat cirrhotic liver induced by CC14. Its phenotype is activated HSCs. The cell line was thawed and cultured in RMPI-1640 medium with bovine serum. After cells proliferated to dense monolayer, they were seeded on coverslides at concentration 2X104/ml. When cells grew confluence, medium was charged to RPMI-1640 without bovine serum. Thereafter, cells were cultured for another 12 hrs in order to synchronize them at G phase. The coverslides were subjected to five groups, namely control, IH764-3 10 mg.L-1, 20 mg.L-1, 30 mg.L-1 and 40 mg.L-1 groups respectively, and IH764-3 was added correspondingly. After 6hrs coverslides were collected and washed three times with PBS, each for five minutes. Then they were fixed in 100% methanol at 4 for thirty minutes in order to stain immunohistochemistrically.Results: The secondary biliary hepatic fibrosis was induced successfully. After BDL 14 days, the livers in model group enlarged obviously and showed up yellow or dark yellow with rough surface and tough texture. There was obtuse edge. Microscopically, liver of model group presentedextensively bile ductular proliferation and ECM deposition res...
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