Perfect Great Fill Soup Compatibility Different Doses Of Cinnamon On Oxidative Damage Rat Dermal Fibroblasts

BackgroundChronic refractory wound is a kind of disease that the process of wound healing is not foreseeable or regular.This kind of wound is characterized by long course of disease,great impact on the appearance,many complications and so on.This disease causes great harm on the quality of patient’s life and work.Domestic and foreign scholars have paid much attention to this disease and carried on much researches.With deep understanding,more and more attention is paid to the impact of oxidative stress on wound healing. Oxidative stress is a state that the imbalance of pro-oxidants and antioxidants induces that more free radicals are produced and/or the ability of antioxidants is decreasing.Excessive oxidative stress can cause tissue damage and result in the delay of wound healing or even long-term healing.Dermal fibroblasts are the major cells in the repaired period of wound healing.Because of oxidative stress in wound healing,lots of reactive oxygen species can make fibroblasts oxidative-damaged and affect their biological behaviors.So dermal fibroblasts can not play the normal function in wound healing.In recent years,there are more and more researches on the functions of fibroblasts in wound healing.In the meantime,the experimental studies on Traditional Chinese Medicine(TCM) and wound healing are developing greatly.The mechanism of TCM on chronic refractory wound can be clarified from the cellular level by the studies on the regulation of TCM on oxidative-damaged fibroblasts.In clinic,TCM have some unique advantages in curing chronic refractory wound.Chronic refractory wound belongs to "yin syndrome ulcer"in TCM.The principle of replenishing should be followed in treatment.The result of treatment will be better to add certain amounts of Chinese herbal medicines of warming-dredging on the basis of Chinese herbal medicines of replenishing. But excessive amounts of Chinese herbal medicines of warming-dredging will play the opposite role in treatment.Therefore,the dose of Chinese herbal medicines of warming-dredging has become an important issue.Shiqudabu(SQDB) is one of the recipe of replenishing and warming-dredging.SQDB is composed of much Chinese herbal medicines of replenishing and a little cinnamon.SQDB is used to cure chronic refractory wound in clinic. In the meantime, experimental studies shows that SQDB has the antioxidative effects.Objective Our objective is to investigate the regulation of Shiqudabu(SQDB) and different doses of cinnamon compatibility on the oxidative-damaged rat dermal fibroblasts and study from the cellular level the mechanism of SQDB and different dose of cinnamon on chronic refractory wound induced by oxidative stress.MethodsRat dermal fibroblasts was got by primary culture. The method of intragastric administration of equivalent dose decoction was employed to prepare5groups of different rat serums,the Normal Control group,the No Cinnamon group,the SQDB group,the Double Cinnamon group and the Four Times Cinnamon group.Use certain concentration of H2O2to damage rat dermal fibroblasts to make the model of oxidative-damaged rat dermal fibroblasts.The serums described above were used to interfere the oxidative-damaged rat dermal fibroblasts and MTT-assay was used to measure the effects of the serums on cell proliferation;PI staining and PI/Annexin V double-staining combined flow cytometry were employed to test the effects on cell cycle and apoptosis;DCFH-DA probe and JC-1probe combined flow cytometry were used to test the effects on the level of reactive oxygen species and mitochondrial membrane potential.Results1.Cell proliferation was significantly inhibited by100-1000umol/L H2O2which had stimulated rat dermal fibroblasts for30minutes. Inhibitory Rate(IR) increased gradually as the increase of the concentration of H2O2.IR was about50%when the concentration of H2O2was500umol/L which was considered as the most appropriate concentration for the following experiments.2.OD value decreased significantly in MTT-assay when500umol/L H2O2had stimulated rat dermal fibroblasts for30minutes.And the proportion of G0/G1period increased significantly while the proportion of S and G2/M period decreased significantly.The rate of apoptosis increased significantly. Mitochondrial membrane potential decreased significantly.And the level of reactive oxygen species increased significantly.3.After24hours of serums treatment,the OD values of the Double Cinnamon group and the Four Times Cinnamon group increased in the group of10%serum concentration. The OD value of the Double Cinnamon group increased significantly in the group of15%serum concentration. After48hours of serums treatment, the OD values of the Double Cinnamon group and the Four Times Cinnamon group increased significantly in the group of5%serum concentration. The OD value of the Double Cinnamon group increased significantly in the group of10%serum concentration.The OD values of the Double Cinnamon group in two timepoints were both the highest in the four groups. 4. After24hours of serums treatment,the proportion of G0/G1period of all four groups decreased significantly,and the proportion of S and G2/M period of all four groups increased significantly. After48hours of serums treatment, the proportion of G0/G1period of the SQDB group,the Double Cinnamon group and the Four Times Cinnamon group decreased,and the proportion of S and G2/M period of the three groups increased. The Proliferation Index of the Double Cinnamon group in two timepoints were both the highest in the four groups.5. After24and48hours of serums treatment,the rate of apoptosis of all four groups decreased significantly. The rate of apoptosis of the Double Cinnamon group in two timepoints were both the lowest in the four groups.6. After24hours of serums treatment, mitochondrial membrane potential of the Double Cinnamon group increased significantly. After48hours of serums treatment,all four groups didn’t have apparent effects on mitochondrial membrane potential.7. After24and48hours of serums treatment,the level of reactive oxygen species of all four groups decreased significantly.ConclusionSQDB and different doses of cinnamon compatibility can protect oxidative-damaged rat dermal fibroblasts by promoting cell proliferation,inhibiting apoptosis and cleaning reactive oxygen species.And the function is more significant to increase the dose of cinnamon on the basis of SQDB.