| ObjectiveDalbergia pinnata is a resin-rich woody vine stem of the legume Dalbergia pinnata(Lour.)Prain,which is warm in nature,bitter and pungent in flavour,and has the effects of dispelling wind and relieving pain,activating blood circulation and astringent sores.Its main treatment is rheumatic paralysis and pain,bruises and injuries,menstrual disorders,and ulcers of the lower limbs.The group’s preliminary research found that the residents of Hainan,Yunnan and other southwestern regions of China often use it to make wine for the treatment of bruises,or crushed and applied to the affected areas for the treatment of burns,wounds,lower limb ulcers,etc..However,although it has a long history of folk medicinal use and precise efficacy,there are no quality standards,simple and crude folk usage,unknown active parts,and unclear mechanism of action,all of which limit the further development and application of Dalbergia pinnata.Based on the concept of "Chinese medicine is the treasure of ancient Chinese science and the key to unlocking the treasury of Chinese civilization" put forward by General Secretary Xi Jinping,in order to fully explore the folk medicinal resources and expand the scope of application of Dalbergia pinnata for the benefit of patients.The group firstly developed the preliminary quality standard of Dalbergia pinnata in the early stage.On this basis,the subject investigated the effects of extracts from different parts of Dalbergia pinnata on the repair of burns,screened out the effective parts,and explored its mode and way of action on the repair of wounds to provide a theoretical basis for the development of Dalbergia pinnata topical drugs.In addition,this project also analyzed the composition of each extracted part of Dalbergia pinnata to provide a theoretical basis for the development of subsequent preparations of Dalbergia pinnata.Methods1.Study on the preparation process and chemical composition of Dalbergia pinnata extractsThe extraction rate was used as an indicator to optimize the best extraction process of the alcoholic extracts of Dalbergia pinnata by investigating the ethanol concentration,extraction time,liquid-to-solid ratio,extraction temperature and number of extractions.Different polar solvent extracts were later applied to prepare different polar parts,and the total flavonoid content of each part was determined by UV spectrophotometry.The chemical composition of each part was analyzed by UPLC-Q/TOF-MS and gas chromatography-mass spectrometry(GC-MS).2.Study on the efficacy and action mechanism of Dalbergia pinnata extracts on burnsSD rats were randomly divided into 10 groups: normal group(NG),model group(MG),positive drug group(PDG),vaseline group(VG),Dalbergia pinnata powder group(DPPG),petroleum ether extract group(PEG),ethyl acetate extract group(EAG),nbutanol alcohol extract group(BAG),water extract group(WG),and fibrous residue group(FRG).A rat burn model was constructed and administered daily after burn injury,and dissected and the skin was taken at 7,14 and 21 days after burn injury.Firstly,the effects of Dalbergia pinnata on the pathological condition of burned skin were investigated by visual observation,calculation of wound contraction rate and H&E staining.The effects on inflammatory factors were examined by ELISA for IL-6 and TNF-α.The changes of TGF-β1 during wound healing were examined by ELISA and immunohistochemical staining.The immunofluorescence staining of α-SMA,CD-31 and immunohistochemical staining of VEGF were used to examine the effect of Dalbergia pinnata on neovascularization.Masson staining was used to examine the effect of Dalbergia pinnata on the formation and arrangement of collagen fibers.Finally,Sirius red staining and immunohistochemical staining of Collagen I and Collagen III were used to examine the effect of Dalbergia pinnata on Collagen I and Collagen III production.3.In vitro study of the active site of Dalbergia pinnata for the treatment of burnsThe effects of petroleum ether extract(PEE)and ethyl acetate extract(EAE)on the proliferation and migration of mouse fibroblasts(NIH-3T3)were investigated.The effect on NIH-3T3 cell proliferation was investigated by CCK-8 method and Ed U staining.The effect on NIH-3T3 cell migration was examined by scratch assay and Transwell migration assay.Results1.Study on the preparation process and chemical composition of Dalbergia pinnata extractsThe best extraction process of the alcoholic extract of Dalbergia pinnata: the ethanol concentration was 80%,the extraction temperature was 30°C,the extraction time was 20 min,the liquid-to-material ratio was 15:1(m L/g),repeating three times.The extraction rate of the alcoholic extract under this condition was(65.32 ±0.82)%,and the total flavonoid content was(24.79 ±0.45)mg/g.6 compounds were identified from the petroleum ether extract,44 compounds from the ethyl acetate extract,16 compounds from the n-butanol alcohol extract,and 12 compounds from the water extract.2.Study on the efficacy and action mechanism of Dalbergia pinnata extracts on burnsWound repair was better in the EAE and PEE treatment groups compared to the model group,and these two extracts were able to inhibit excessive inflammatory factor expression,promote neovascularization,and promote collagen fiber formation and orderly arrangement.In addition,the ratio of Collagen I/Collagen III was lower in the EAE and PEE treatment groups,suggesting the possibility of reduced scarring.Finally,EAE and PEE may promote wound repair by upregulating the expression of early TGF-β1.3.In vitro study of the active site of Dalbergia pinnata for the treatment of burnsCCK-8 method and EdU staining results showed that 2,4 and 8 μg/mL of PEE solution and 0.5,1 and 2 μg/m L of EAE solution were able to promote the proliferation of NIH-3T3.The results of scratch and Transwell migration assays showed that 2 μg/m L of PEE solution and 0.5 μg/m L of EAE solution could promote NIH-3T3 cell migration.In conclusion,PEE and EAE could act directly on fibroblasts to promote their proliferation and migration.ConclusionThe above-mentioned studies suggested that EAE and PEE could promote wound repair and might reduce scarring,and their mechanism of accelerated wound repair might be related to the regulation of TGF-β1 secretion.The main component of PEE is elemicin,and EAE contains formononetin,liquiritigenin,butin,and isoliquiritigenin.Their significant effects may be related to the antibacterial,anti-inflammatory,and antioxidant activities of these components related. |
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