The Influence And Prevention In Biejiajian Pills And Rosiglitazone Drugs' Serum To TGF β1Stimulating Rat Hepatic Stellate Cell

Cirrhosis (cirrhosis of liver) is a chronic, progressive, diffuse liverdisease, caused by the various causes liver cell diffuse necrosis, regeneration,induce fibrous connective tissue hyperplasia, leading to a structure destroy andfalse flocculus lobules reconstruction, hence to cirrhosis of the liver.Cause liver cirrhosis for many reasons, different regions of the maincause is not identical also, in Europe and the alcoholic cirrhosis is givenpriority to, our country to hepatitis cirrhosis see most. Worldwide, livercirrhosis has become the second only to cardio-cerebrovascular diseases andmalignancies major disease, great harm to human beings.Liver fibrosis (hepatic fibrosis, HF) is a chronic liver disease commondevelopment basis pathological changes, also is the chronic hepatitis andcirrhosis of the important link of the further development. Liver fiberorganization of excessive deposition and the extracellular matrix (ECM,extracellular matrix composite degradation is greater than the causes of theformation of liver fibrosis, on the one hand, the hepatic stellate cells (hepaticstellate cells, HSC) activation and to muscle into fiber appearance cell and intothe transformation of fiber cells, resulting in a large ECM synthesis, on theother hand the degradation of ECM reduction in the synthesis and degradationof promoting the formation of liver fibrosis balance.So is liver cirrhosis of the liver fibrosis early and inevitable stage, ifcause persist, liver fibrosis will gradually increase, liver lobules and bloodvessels and gradually were rebuilt, make the normal structure of liver damage,central vein area and collect abbacy occurrence interval, and false flocculus toform, development as the irreversible cirrhosis of the liver.TGF-β1is the most important cause of found fibrosis cell factors, is setup HSC liver fibrosis and Bridges. When all kinds of pathogenic factors continuing role in the liver, through the complex mechanism activate HSC,thus cause liver fibrosis and cirrhosis of the liver. The artificial activationprocess can be divided into two stages: start-up phase and continuousactivation stage. Start early stage representative gene activation and phenotypeof change, make of cell factors and HSC comes from the cell factorsstimulation including lipid peroxide and ECM change produce the response;Continuous activation stage mainly by the damage of the local generation cellfactors stimulate, maintain the phenotype of activation and fibrosis formation.TGF-β1can promote HSC proliferation, and promote the secretion Ⅰ,expression and HSC Ⅲ, Ⅳ collagen type, fiber connection protein, cellsticking element, and promoting the via autocrine TGF-β1own expression,another can inhibit collagen enzyme and substrate decomposition vegetableetc MMP and fibrinolytic enzyme activated the expression of the thing, andpromote HSC and endothelial cells express fibrinolytic enzyme activated thething inhibitory factor and TIMP. Therefore, TGF-β1in liver fibrosisoccurrence, development process is important in beginning with continuousactivation function and action, as interference TGF-β1target anti fibrosistreatment established the theory basis.Participate in TGF-β signal transmission of related proteins in cellscalled Smad signal protein family, for TGF-β downstream signaling moleculescollectively. Among them Smad2, Smad3, Smad4, Smad7participation TGF-βsignal transduction pathways. Smad7is TGF β/Smad-signalling pathways ofthe main inhibitory control protein, TGF-β1to signal transduction pathwaysof feedback and cross regulation plays a key role, therefore, the expression ofthe Smad7regulation for the prevention and control of liver fibrosis offers anew way.Western medicine rosiglitazone for PPAR γ synthesis ligands, for PPAR γagonists, PPAR γ activation studies confirm that can inhibit the ECM mRNAexpressions, make ECM synthesis reduce, PPAR γ can inhibit TGF-β1activation HSC access, promote HSC apoptosis, reversal fibrosis. In Chinesemedicine treatment of liver fibrosis also obtained a certain result, traditional Chinese medicine has many link, multi-level and the comprehensivepharmacology function targets, and it has good prevention of liver fibrosis andto the human body low toxicity reaction etc. Characteristics. Turtle shell Friedpills before a series of research that have inhibit collagen synthesis and HSCactivation proliferation, improve the action of liver function. But Chineseherbal compound contains many impurities, such as direct join in vitroreaction system, its physical and chemical factors nonspecific affect theexperiment. So in the study, the serum pharmacology method of normal ratsextraction drugs in serum experiments.Objective:Explore different concentration turtle shell Fried pill to drug serumTGF-β1exciting rat hepatic stellate cells (HSC) on the ECM synthesis andthe influence of the activation HSC proliferation, TGF-β1/smad signaltransduction pathways, and the results and action mechanism and westernmedicine compared rosiglitazone.Methods:1A serum pharmacology rat drug serum: configuration of TGF-β1concentration of droperidol5μg,/L DMEM medium. Were preparedcontaining2.5%and7.5%,12.5%A Pill Drug serum concentration of TGF-β1/L DMEM medium droperidol5μg, respectively, as traditional Chinesemedicine with low, medium, high prevention group intervention medium;preparation containing7.5%Rogge rosiglitazone drug serum, TGF-β1concentration of droperidol5μg, medium/L DMEM medium as a westerndrug prevention group intervention; preparation containing7.5%normal ratserum medium as a control group intervention medium; preparation containing7.5%normal rat serum, TGF-β1concentration of droperidol5μg,/L DMEMmedium as a model group intervention medium.2drugs in vitro culture and HSC serum index detection: the in vitroculture technology application HSC cell lines, in37℃cell culture in the box,stay in DMEM medium stick in the cell wall, in serum-free with DMEMsynchronization training24h, then change to the intervention with medium intervention cells24h, of the indicator detection:(1) spectrophotometry(optical density, OD): MTT method (2) ColIV, LN: radiation immune method(3) TIMP-1mRNA, MMP-9mRNA, Smad7mRNA: the transcriptionpolymerase league reaction (reverse transcription-polymerase chain reaction,RT-PCR)Results:1Results show that the MTT: TGF-β1can significantly improve HSCsproliferation (P <0.05), different doses turtle shell Fried pills and rosiglitazonedrugs are obviously down-regulated serum TGF-β1exciting HSCsOD value(P <0.05), high dose group and control group OD value difference wasstatistically significant (P﹥0.05).2Put from the result shows: the TGF-β1can increase ColIV, LNsynthesis (P <0.05), and the model group, each drug intervention groupcompared ColIV, LN content decreased (P <0.05), but compared with thecontrol group still have difference (P <0.05).3RT-PCR results show that: TGF-β1improved obviously TIMP-1mRNA expression (P <0.05), reduce MMP-9mRNA, Smad7mRNAexpression (P <0.05). Compared with model group, each drug interventiongroup all can make TIMP-1mRNA express to decrease (P <0.05), thatMMP-9mRNA, Smad7mRNA express increased (P <0.05), but comparedwith the control group still have difference (P <0.05).4turtle shell Fried pill inhibition of cell proliferation, collagen synthesis,TIMP-1mRNA, MMP-9mRNA, Smad7mRNA expression is the role of thedose-response (P <0.05).5turtle shell Fried pill high dose group of inhibiting cell proliferation,collagen synthesis, TIMP-1mRNA, MMP-9mRNA, Smad7mRNAexpression is better than the role of rosiglitazone (P <0.05), the dosage groupwith rosiglitazone is no statistical difference (P <0.05), low dose group oflower than the role in the rosiglitazone group (P <0.05).Conclusions:1TGF-β1can significantly improve HSCs proliferation activation and ColIV, LN synthesis secretion, and its function may be smad7mRNAexpression through improving the lower MMP-9mRNA, smad7mRNAexpression, promote the ECM synthesis and activate TGF-β1/smad signaltransduction pathways to realize.2rosiglitazone and different doses turtle shell Fried pills inhibits TGFserum all drugs-β1the role, but both are cannot make HSCs state back toTGF-β1stimulate level before3turtle shell Fried pill role within a certain range is dose dependent.Turtle shell Fried pill high dose group of effect is better than the rosiglitazone,turtle shell Fried of the low-dose pill in the role of the poor in the rosiglitazonegroup, turtle shell Fried pill dose group of in the rosiglitazone group andsurfactant therapy.