The Effect Of Methyl Ferulic Acid On Proliferation Of Human Hepatic Stellate Cells And The Expression Of α-SMA And TypeⅠ Collagen

Hepatic fibrosis(hepatic fibrosis, HF) is a pathological change that causedby variety of chronic liver damages, inflammation and necrosis of the liverparenchyma persistently or recurrently, fiber connective tissue degradation isrelatively insufficient in the liver, making lots of the fiber connective tissuehyperplasia, leading to the phenomenon of the extracellular matrix componentsexcessive unusually sedimentary, thus forming to of liver fibrosis, further gettingto development of liver cirrhosis. The main reasons of liver fibrosis includingviral hepatitis, alcoholic and biliary obstruction and so on. If we can giveeffective treatment，can we relieve the degree of liver fibrosis, make it changeover. Hepatic stellate cell is the main cell of the formation of liver fibrosis, andis the main source of extracellular matrix element. Under normal circumstances,HSC is mostly storing vitamin A that is proliferation slightly, compounding asmall amount of collagen. However，Vitamin A disappeared in Hepatic stellatecells as well as turning into Muscle fiber sample cells after the liver damage,that is the activation of HSC. The activation of HSC can synthesize and secret ECM，and lead to the ECM excessively silting up in the liver，and expressingα-SMA.Transforming growth factor beta1can promote HSC into Muscle fibersample cells(Muscle fibroblasts), increasing the generation of extracellularmatrix and reducing the degradation of extracellular matrix, inhibiting liver cellregeneration and induction of apoptosis. TGF-β1is considered to be the mostfibrosis factor that played an important role in the development of theoccurrence of liver fibrosis.Chronic liver disease that induced by a variety of reason seriously threat tothe health of human in our country. Furthermore, to seek for effective anti-liverfibrosis drug is a hotspot in current research. Nowadays lots of research takehepatic stellate cells as a central part about anti-fibrosis, but there is no specifictreatment. Securidaca inappendiculata Hassk is the national characteristics ofChinese herbal medicine in guang xi province. It is people said that it is oftenused in the treatment of acute jaundice hepatitis C. Our research team collectedMethyl ferulic acid after extraction and separation and purification from bark ofsecuridaca inappendiculata Hassk, discussing the effect of MFA in the process ofhepatic stellate cell activation and proliferation and its molecular mechanisms ofits function. It is a main significance to develop the drug to resistance the liverfibrosis.objective： To discuss MFA impact on human hepatic stellate cellsproliferation, the effect of MFA on the expression of mRNA level of α-SMA andtype Ⅰ collagenas well as the protein expression of α-SMA, and its possible mechanism.Methods: Human hepatic stellate cells(HSC-LX2) as the research object,MTT assay was used to test MFA impact on proliferation of human hepaticstellate cells and detect the influence of TGF-β1on the proliferation of HSC-LX2when its concentration is5ng/ml;TGF-β1stimulated of human hepaticstellate cells as oxidative stress model of liver fibrosis. And based on applicationof different doses of methyl ferulic acid intervene groups of cells, HSC-LX2cells were stimulated with MFA in different doses, he expression of mRNA levelof α-SMA and type Ⅰc ollagenwere evaluated by RT-PCR. The proteinexpression of α-SMA were evaluated by Western blot.Results:①MTT results showed that MFA can suppress the proliferation ofHSC-LX2after48h(P<0.05). Compared with normal control group，TGF-beta1can obviously promote the proliferation of HSC-LX2when its concentrationis5ng/ml(P<0.05).②R T-PCR results showed that under the stimulation of TGF-beta1,MFA can reduce the expression of mRNA level of α-SMA and typeⅠ collagenafter48h(P<0.05), and with a low concentration of MFA (5μg/ml) ismost obvious, high and middle dose of MFA（10μg/ml、5μg/ml）can alsoinhibition expression of HSC-LX2mRNA level of α-SMA and type Ⅰcollagen(P<0.05).③Western blot asssay releaved that, low concentration ofMFA(5μg/ml) is able to bring down the protein expression of HSC-LX2α-SMAsignificant (P<0.01). Model group was stimulated TGF-beta1compared， medium and low dosage MFA also can inhibit the protein expression ofHSC-LX2α-SMA, but there is no obvious compared with high concentrationgroup.Conclusion：①MFA can restrain the proliferation of human hepatic stellatecells.②MFA pretreatment TGF-beta1provoked human hepatic stellate cells,that can reduce the mRNA expression of a-SMA and type Ⅰcollagen as well asprotein expression of a-SMA，thus infer that MFA can inhibit proliferation andactivation of HSC，inhibit the occurrence and development of liver fibrosis.