Study On The Active Substance Mechanism For The Anti-chronic Nephritis Effect Of Rostelluaria Procumbens(L.)

Chronic kidney,a multiple and serious disease in modern clinical,was characterized by its great incidence,low cure rate,high cost of treatment,longer treatment time and poor quality of life for patients.Up till the present moment,most drugs used to treat chronic nephritis have severe toxicity and the therapeutic effect was unsatisfactory.Just because of this,the work of develop a new effective and lower toxicity drugs is imminent.Herba Justiciae is the whole dried plant of Rostelluaria Procumbens(Lin.)Nees and included in the Chinese Pharmacopoeia(1977 edition,a).Traditional Chinese Medicine take advantage of function such as Qing re jie du,Li shi xiao ji,Huo xue zhi tong to cure cold and fever,cough,swelling and pain in throat and so on.In the treatment of chronic nephritis,Traditional Chinese Medicine opt in Herba Justiciae based on the original Chinese medicine compound and utilize its virtue of Li shi xiao ji to relieve edema of chronic nephritis.Even though it is widely used,but its mechanism is unclear and further research is still needed.In this thesis,Lipopolysaccharides(LPS)was used to induce mesangial cells HBZY-1 to construct the chronic nephritis model.By the used of system solvent separation method,Rostelluaria Procumbens(Lin.)Nees’s et hanol extract was divided into petroleum ether,ethyl acetate,n-Butanol and aqueous solution of four fraction,and then the four parts of pharmacological activity were filtered by the following way.The influence of drugs on mesangial cell proliferation and Cytokines(IL-beta,IL-6,IL-1 TNF alpha)secretion was evaluated by MTT and ELISA kit,respectively.The results showed that petroleum ether,ethyl acetate parts can inhibit the proliferation of mesangial cells and the release of cytokines to a certain degree.According to the results of part screen,the petroleum ether was selected to further separate,it was separated into four parts Labeled as A,B,C,D and used the same method with parts screening for active filter.The experimental data show that A and B can inhibit the proliferation of mesangial cells and the release of cytokines to a certain degree.In order to isolate A,B two parts and identify structures of the compounds,Silica gel column chromatography,thinlayerchromatography(TLC),high-performance liquid chromatograp hy(HPLC),Ultraviolet-Visualabsorptionspectrometry(UV),~1H-nuclear resonance(~1H-NMR)and so on were adopted.Eight compounds were isolated from A,B and identified as Justicidin A,neojusticin A,Justicidin B,neojusticin B,stigmasterol andβ-sitosterol.Activity screening method of compounds is the same as the method of part screening.The date suggest that only Justicidin A has a less obvious effect.It can inhibit the proliferation of mesangial cells and the release of cytokines to a certain degree.To further verify the pharmacology activity of each parts,each paragraph and each monomeric compounds,western blot was chosen to qualitative and quantitative analysis of proteins related to TLR4/NF-κB signal pathway,such as TLR4,MyD88,NF-κB and phosphorylase-NF-κB.Western blot data shows that the results were consistent with the results of ELISA.The level of protein was down regulated after the medication reacted on HBZY-1 and it is indicated that Rostelluaria Procumbens(Lin.)Nees in the treatment of chronic nephritis may be through the NF-κB pathway to adjust the body function and achieve the purpose of cure.