Study On Anti-Inflammatory Activity And Anti-Mastitis Effect Of Mouse Of "Ruyan Ning" Water Extract

Due to the difficulty and high cost of developing new chemical anti-inflammatory drugs and the slow updating speed of chemical anti-inflammatory drugs in recent years,many domestic and foreign scholars have surrendered their attention to traditional Chinese medicine anti-inflammatory drugs.“Ruyan Ning”formula is a new formula for the treatment of inflammatory diseases,which is based on the compatibility principle of traditional Chinese medicine and has been continuously improved by our research group after years of clinical experience.Previous studies have shown that the single Chinese herbal medicine in this prescription has certain anti-inflammatory effects,so it is necessary to study the anti-inflammatory effects of this prescription and fill in the gaps in previous studies.Objective:In this study,mice were used as the main test animals,and“Ruyan Ning”water extract was used as the main test drug.The water extract was given to mice at different dose concentrations by gavage to investigate its anti-inflammatory activity and anti-mastitis effect in mice,and to provide a theoretical basis for its clinical application and further research.Methods:The experiments were carried out immediately after“Ruyan Ning”water extrac twas administered intragastric at low（4 mg/g）,medium（8 mg/g）and high（16 mg/g）doses.（1）Ear swelling induced by xylene,peritoneal capillary permeability increased by acetic acid,foot swelling induced by carrageenan,granulation swelling induced by cotton balls,and serum cytokines TNF-αand IL-1βcontents of mice were detected.The experiment lasted for 10 days.At the end of the experiment,ear weight,abdominal lotion D_{590 nm},foot weight,cotton granuloma weight and serum contents of TNF-αand IL-1βwere measured.（2）Mice were intraperitoneally injected with 4×10⁸CFU/Staphylococcus aureus.2 h after the modeling,each drug treatment group was given the drug,once a day for 7 days,during which the death and survival status of mice were recorded every day.At the end of the experiment,blood,heart,liver,spleen,lung and kidney of each group were collected.Serum was used to detect TNF-αand IL-6,and histopathological sections of each tissue were made by HE staining.（3）The UC model was constructed by feeding 3%DSS solution for 7 days.After successful modeling,the drug treatment group was administered once a day for 10 days,during which stool characteristics and living conditions of mice were observed every day.At the end of the experiment,the colon tissues of each group were collected for pathological observation and the contents of IL-1β,IL-17,TNF-α,IFN-γ,PPARγ,GSH-Px,SOD,MPO and MDA in the colon tissues.（4）Mice were treated with LPS for mastitis modeling.2 h after the modeling,the mice were administered once a day for 7 days.At the end of the experiment,mammary gland tissues of mice in each group were collected,part of which were used for histopathological observation,and part of which were used to detect the contents of TNF-α,IL-1β,IL-6 and MPO in mammary gland.Results:（1）The results showed that"Ruyan Ning"water extract in each dose group had very significant anti-inflammatory effects on ear swelling caused by xylene,peritoneal capillary permeability increase caused by acetic acid,foot swelling caused by carrageenan and granulation swelling caused by cotton balls（P<0.01）.The ELISA results showed that compared with the BC group,the serum levels of RYN-M,RYN-H groups all had a highly significant decrease in serum IL-1βcontent（P<0.01）,and serum TNF-αcontent was significantly lower in the RYN-M group（P<0.05）and highly significantly lower in the RYN-H group（P<0.01）.（2）The results showed that RYN-M and RYN-H could significantly improve the survival rate of model mice（P<0.01）.HE staining of liver,lung and kidney tissues of mice showed that the structural integrity and inflammatory infiltration of model mice were improved in each dose group of“Ruyan Ning”water extract.ELISA results showed that the contents of TNF-αand IL-6 in serum of model mice were decreased in each dose group of“Ruyan Ning”water extract（P<0.01）.（3）The results showed that DAI of mice in Model group and“Ruyan Ning”water extract groups was significantly increased（P<0.01）,and colon length was significantly shortened（P<0.01）.In addition,each dose group of“Ruyan Ning”water extract can improve the damage degree and inflammatory infiltrating state of colon tissue in UC mice model induced by DSS.ELISA results showed that,compared with Model group,the detected cytokines in each dose group of“Ruyan Ning”water extract had a very significant recovery trend（P<0.01）.（4）The results showed that each dose group of“Ruyan Ning”water extract could improve the structure of breast tissue and relieve inflammatory infiltration.ELISA results showed that the contents of cytokines TNF-α,IL-1β,IL-6 and MPO in mammary gland tissue of model mice were significantly decreased in each dose group of“Ruyan Ning”water extract（P<0.01）.Conclusion:“Ruyan Ning”water extrac has good anti-inflammatory effect and has positive modulating effect on the mouse model of inflammation caused by Staphylococcus aureus infection,mouse UC model and LPS-induced mastitis model in mice,among which the high dose group（16 mg/g）of“Ruyan Ning”water extrac.