Study The Strong Quality Evaluation Index Of Radix Millettia Championi And Somg Of Its Efficacy

Objective: To investigate the contents of Maackiain and Formononetin from Radix Milledttia Speciosa and the anti-inflammatory,improve immunity and protective effect of ALI of the total flavonoids of Radix Millettia Championi(FRMC),and explore in the effect of the difference between FRMC and the total flavonoids of Radix Millettia Speciosa(FRMS),For Radix Millettia Championi in anti-inflammatory,improve immunity and ALI can be used as a Radix Millettia Speciosa alternative or supplements and medicinal value of the development and application of experimental basis and theoretical basis.Methods: 1.To determine the content of Maackiain and Formononetin in Radix Millettia Championiwere Radix Milledttia Speciosa,Maackiain and Formononetinused were used as the standers,and the mobile phase consisted of nitrilewith Welchrom-C18(4.6 mm × 250 mm,5?m)column.The detection wavelengths were 248 nm and 309 nm.The contents of Maackiain and Formononetinused in Radix Millettia Championi and Radix MilledttiaSpeciosa were determined and compared.2.To investigate the anti-inflammatory and improve immunity of FRMC,several inflammatory models,including the swelling of ear induced by xylene in mice,the swelling of paw induced by carrageenan in mice,the increasing of capillary permeability in mice' s abdominal cavity by acetic acid,the cotton-pellet granuloma of mice,and cyclophosphamide-induced immunosuppression model in mice were used to explore the anti-inflammatory and immune effects of FRMC and observe the similarities and differences of anti-inflammatory and improve immunity of FRMC and FRMS.3.To investigate the protective effects of FRMC and FRMS on ALI,the number of white blood cells(WBC)and the total protein(Pro)in the bron choalveolar lavage fluid(BALF),the pulmonary edema coefficient,Superoxide Dismutase(SOD)and Malondialdehyde(MDA)in lung tissue were measured.The levels of tumor necrosis factor-alpha(TNF-?),interleukin 6(IL-6)and interleukin-1 beta(IL-1?)in the lung tissue were detected by enzyme-linked immunosorbent assay(ELISA).The morphological changes of lung tissue were observed by hematoxylin-eosin staining(HE).4.To investigate the mechanism of protection of FRMC and FRMS on ALI,the expression of Nuclear factor ?B p65(NF-?B p65)protein in lung tissue of mice was detected by immunohistochemistry,and the expression of IL-6,TNF-?,NF-?B p65 and Toll-like receptors 4(TLR4)mRNA in lung tissue of mice were detected by real-time fluorescent quantitative Polymerase ChainReaction(PCR).Results:1.The distribution trend of Maackiain and Formononet: root of Radix Millettia Championi are higher than Millettia Speciosa(P<0.05).2.FRMC had significant inhibitory action on ear edema induced by xylene in mice and paw edema caused by carrageen in mice(P<0.05 or P<0.01).FRMC also significantly decreased the peroneal of capillaries permeability and the proliferation of granuloma caused by implantation cotton pellet in mice(P<0.05 or P<0.01).the middle and high dose of FRMC and FRMS groups could improve the thymus index,clearance of charcoal particles(K),phagocytic indices(?)and delay type hypersensitivity(DTH)of immunosuppressed mice(P<0.05 or P<0.01).Among those indexs,low and middle dose of FRMC had a stronger effect than FRMS in xylene-induced ear swelling and clearance of charcoal particles(P<0.05),while FRMC had weaker effect than FRMS in inhibition of the cotton-pellet granuloma(P<0.05).3.Compared with the model control group,FRMC and FRMS could reduce pulmonary edema coefficien,Pro and capillary permeability(P<0.05 or P<0.01),decrease the number of WBC in BALF at different levels(P <0.05 or P<0.01).FRMC and FRMS also could improve the activity of SOD and reduce the amount of MDA in lung tissue at different levels(P <0.05 or P <0.01).The content of TNF-?,IL-6 and IL-1? in lung tissue were also decreased in different levels by FRMC and FRMS.Among them,the high dose group of FRMC was weaker than FRMS in improving the activity of SOD(P< 0.01),and the middleand high dose group of FRMC was weaker than FRMS in reducing the content of MDA(P < 0.05).In the remaining indicators,the effects of FRMC and FRMS were similar.Pathological examination showed that,FRMC and FRMS groups could protect ALI mice by improving the inflammatory cells,erythrocytes and other exudates,pulmonary interstitial hyperplasia,pulmonary edema.4.Compared with the model control group,FRMC and FRMS could reduce the expression of NF-?B p65 protein in lung tissue(P<0.01).FRMC and FRMS also could inhibit the expression of TLR4,NF-?B P65,IL-6 and TNF-?mRNA in different level(P <0.05 or P <0.01).Among them,the low dose of FRMC was stronger than FRMS in reducing NF-?B p65 protein,NF-?B p65 and TLR4 mRNA,and the high dose of FRMC were stronger than FRMS in reducing IL-6 m RNA.In the remaining indicators,The effect of FRMC and FRMS were similar(P> 0.05).Conclusions: 1.The HPLC method can be used for the determination of the content of Maackiain and Formononetin.2.FRMC has a good anti-inflammatory and improving immune function.FRMC is stronger than FRMS in the inhibition of the ear swelling,and weaker than FRMS in the inhibitory effect in the cotton-pellet granuloma and and improve immune function..3.FRMC and FRMS have protective effects on LPS-induced ALI,and The protective mechanisms of FRMC and FRMS are through the interference of TLR4/NF-?B pathway,then expression and generation ofinhibiting the inflammatory factor.