Study On The Activity Of Ligia Exotica Roux And Its Symbiotic Microorganisms In The Treatment Of Soft Tissue Injury

Objective:The activities of marine traditional Chinese medicine Ligia exotica Roux and its symbiotic microorganisms in the treatment of muscle injuries were studied,and the diversity of symbiotic microorganisms of Ligia exotica Roux was studied.By analyzing the effects of crude extracts and different parts of Ligia exotica Roux on soft tissue injury model rats,the community structure and diversity of symbiotic microorganisms of Ligia exotica Roux and the effects of crude extracts and microbial metabolites of Ligia exotica Roux on inflammatory cells were analyzed.To provide a research basis for the discovery of compounds for the treatment of tendon injuries,so as to promote the application of marine traditional Chinese medicine in the treatment of tendon injuries.Methods:1.Study on the effect of crude extract of Ligia exotica Roux and its different parts in the treatment of tendon injury.According to the requirements of pharmacological experiment,264 male SD rats of SPF grade were selected and divided into 11 groups with 24 rats in each group:blank group,model group,positive drug group,high and low dose of total extract of Ligia exotica Roux,high and low dose of ethyl acetate,high and low dose of n-butanol,high and low dose of water residue.The soft tissue injury model rats were made by gravity beating.24 hours after the success of the model,the drug was given,and the corresponding drugs were smeared respectively in the treatment group,and normal saline was smeared in the blank control group and the model group once a day for 5consecutive days.After 1,3 and 5 days of administration,8 rats in each group were random Ly selected to hemorheology and PGE₂in the injured tissue.2.Ligia exotica Roux can cultivate symbiotic microbial diversity.Ligia exotica Roux were collected from different sea areas and different seasons,and six different isolation media were selected to isolate symbiotic bacteria.The bacteria were isolated and purified by pure culture,and the species were identified by 16S r RNA and the strains were analyzed by Veeny.3.Study on the activity of alcohol extract of Ligia exotica Roux and its microbial fermentation products to cellular inflammationThe cytotoxicity was detected by MTT method,and the concentration suitable for cell growth was determined.The inflammatory model of macrophages in RAW264.7 mice was induced by LPS,and the effects of alcohol extracts and metabolites of Ligia exotica Roux on cell activity and NO secretion were detected.Results:1.Results of alcohol extract of Ligia exotica Roux and its different parts in the treatment of soft tissue injury:（1）From the detection results of PGE₂and IL-1β,High dose of Ligia exotica Roux and high dose of n-butanol could reduce the content of PGE₂and IL-1βin injury tissue.（2）High dose of Ligia exotica Roux extract group,ethyl acetate high dose group and n-butanol high dose group could reduce the content of MDA in injured tissue and increase the content of SOD in injured tissue.（3）High dose of Ligia exotica Roux extract,high dose of ethyl acetate and high dose of n-butanol could improve the whole blood viscosity of soft tissue injury model rats,and the therapeutic effects of ethyl acetate and n-butanol increased in a dose-dependent manner on the indexes of fibrinogen and plasma viscosity after 5 days.2.Results of culturable symbiotic microbial diversity of Ligia exotica Roux:（1）19,6 and 8 culturable bacteria were isolated from Ligia exotica Roux in Qinzhou,Fangchenggang and Beihai sea area of Guangxi.Vibrionaceae was the dominant bacteriaceae of Qinzhou,Fangchenggang in this experiment.Rhodobacteraceae was the dominant bacteriaceae community in Beihai.（2）The species number and diversity richness of bacteria isolated from Ligia exotica Roux in Qinzhou sea area.And one common bacteria can be cultured in three different sea areas.Compared with the sea areas of Qinzhou and Fangchenggang,there were two culturable bacterias;whereas Staphylococcus was shared between the bacterial of Qinzhou and Beihai.（3）The number of bacteria isolated from R₂A medium was the largest,followed by M9 medium.3.Study on the activity of alcohol extract of Ligia exotica Roux and its microbial fermentation products to cellular inflammation:（1）In addition to the inhibitory effect of GXIMD07824 metabolites on cell proliferation,the metabolites of Ligia exotica Roux drug groups and other bacteria fermentation groups at the concentration of 100μg/m L had no obvious toxicity to cells,but some could promote cell proliferation.The metabolites of GXIMD07953 were obtained at the concentration of 100μg/m L,200μg/m increased cell proliferation in a concentration dependent manner.（2）In the drug of Ligia exotica Roux group and GXIMD07836,GXIMD07918 and GXIMD07913 metabolites,the content of no was decreased,while GXIMD07922,GXIMD07904 and GXIMD07972metabolites decreased the content of NO released by inflammatory cells in a dose-dependent manner.Conclusions:1.Ethyl acetate and n-butanol of Ligia exotica Roux can improve and repair the damaged tissues of soft tissue injury model rats from different angles,and achieve the effect of treatment and improvement.Provide preliminary research basis for the material basis of the follow-up directional excavation of Ligia exotica Roux for the treatment of soft tissue injury.2.In summer,Qinzhou sea area was the best sampling place and time of Ligia exotica Roux to explore microbial species resources,and further explore the pharmacodynamic activity of microorganisms,so as to increase the utilization of marine traditional Chinese medicine Ligia exotica Roux and explore the medicinal value of microorganisms.3.There were active components in the fermentation products of six strains and the drug of Ligia exotica Roux groups,which can promote the growth and proliferation of the cells.After LPS was used to induce cells,it was found that some strains had the effect of reducing NO content.