Study On The Mechanism Of The Treatment Of Burn Wound By Accumulated Snow And Grass

Research Background:With the increasing in the aging population,the incidence of chronic wounds such as diabetic ulcers,radioactive ulcers,and pressure ulcers has increased significantly.Therefore,wound healing has become one of the focuses of modern medical research.The asiaticoside cream ointment is extracted from the whole plant of Centella asiatica and has the function of promoting wound healing.It is used clinically to treat surgical wounds,burns,trauma,and keloids.The exact mechanism of asiaticoside in promoting wound healing has not yet been clarified.In particular,the molecular mechanism that promotes the healing of burn wounds has become a hot topic for scholarsObjective:In order to further study the molecular mechanism of asiaticoside in promoting wound healing,the effect of asiaticoside on the expression of CyclinB1,CyclinC,CyclinD1 and proliferating cell nuclear antigen(PCNA)during the healing process of burn wounds was studied to explore its effect on wound healing.The mechanism of healing,and by studying the effect of asiaticoside on inflammatory factors and NF-?B signaling pathway in burn wound tissue,explore the mechanism of action of inhibition of infl ammation.Methods:Part one:72 animal models with deep ?°scald were established.The wounds of rats were divided into two groups according to different treatment methods..These were the asiaticoside drug group and the control group,and the wound was observed and recorded.In the healing condition,the wound specimens on the first,third,fifth,seventh,ninth and fourteenth days after injury were stained with HE and immunohistochemistry to observe the cell proliferation and the expression of CyclinBl,CyclinC and PCNA.Paired data t-test was used for the two groups,and P<0.05 was considered statistically significant.Part two:Wistar rat deep scald animal model of wound tissue cells were taken for cell culture,and the resulting cells were taken and digested.The cells were seeded in a 96-well plate at a density of 2×104-3×104 cells/mL for 12 hours.Randomly divided into blank group(without drugs),the final concentration of asiaticoside 10-7,10-6 and 10-5mol/mL three dose culture groups,a total of 4 groups,each group of 8 holes.The effect of asiaticoside on scald epithelial tissue cell proliferation was detected by MTT assay.Detection of the number of macrophages in wound epithelial tissue by immunohistochemistry;The TNF-a in the supernatant was detected by ELISA;The expression of a,IL-1 and IL-10.The expression of NFkB P65 in wound tissue was detected by Western blot method.The activation of NF-KB was detected by laser confocal microscopy(translocation of major subunit p65 into the nucleus)Results:Part one:? Gross observation:Deep ?° scald wounds in Wistar rats.The asiaticoside treatment group completely healed within(17.40±3.15)days.In the control group,except one wound healed 25 days after wound infection,the other wounds were approximately.(22.17±4.02)d healed,there was a statistically significant difference between the two groups(P<0.05)·? HE staining:At the 3th day,the asiaticoside-treated group showed obvious hyperplasia of epidermal cells,and the control group showed significant proliferation on the 7th day.?The results of immunohistochemical staining of wound specimens on the 1st,3rd,7th,11th,14th,and 17th day after injury showed that the expression of CyclinBl and PCNA were significantly different between the two groups(P<0.05),while the expression of CyclinC was not different.Statistical significance(P>0.05).Part two:?The cell growth was detected by MTT assay:compared with the blank group,the three dose groups of Asiaticoside had no significant effect on cell proliferation(P>0.05),indicating that the concentration of asiaticoside on epithelial cells No cytotoxic effect,with a certain degree of safety.?The number of invasive macrophages in the third,seventh,eleventh and fourteenth day of the treatment group was less than that in the control group.?ELISA to detect the expression of TNF-?,IL-1 and IL-10 in the supernatant:compared with the blank group,each dose of asiaticoside inhibited the excessive expression of TNF-a and IL-1 in different degrees(P<0.05),and the concentration of TNF-a and IL-1 in the asiatic acid group showed a decreasing trend with increasing concentration;the expression of IL-10 in asiaticoside dose group was significantly higher than the blank group.The increase in dose is on the rise.?The expression of NFkBP65 protein in the tissue of wound 7 days after injury was detected by WB method.?The nuclear translocation of NF-KB was observed under laser confocal microscope:the percentage of positive cells in the blank group was(80.20±8.11)%,and the red-stained p65 subunit was mainly concentrated in the nucleus,indicating that the p65 subunit was transported into the nucleus;The percentage of cells translocated into the nucleus of p65 in each dose group was significantly reduced in a dose-dependent manner,and the difference between the four groups was statistically significant(F=24.172,P<0.001).Compared with the blank group,asiaticoside There was significant difference between the 10-6moL/mL group(t=3.535,P=0.001)and Asiaticoside 10-5 moL/mL group(t=6.675,P<0.001)Conclusion:?Asiaticoside can promote the healing of burn wounds by increasing the expression of CyclinBl and PCNA.Asiaticoside can promote the healing of burn wounds by increasing the expression of CyclinB1 and PCNA.To understand the mechanism of asiaticoside in promoting burn wound healing can be combined with other drugs and the effects of several drugs can be superimposed.The different mechanisms of promoting wound healing can be fully utilized to accelerate the wound healing as a whole and shorten the healing time.?Asiaticoside can inhibit overexpression of inflammatory response by inhibiting the nuclear translocation of NF-KB major subunit p65 protein.In order to understand the mechanism of asiaticoside inhibiting inflammatory reaction,combined with the time characteristics of burn wound inflammatory reaction,the use of asiaticoside during the inflammatory period of burn wound can reduce the inflammatory reaction of wound,promote wound healing and shorten the healing time.?asiaticoside has practical application in the treatment of burn wounds.It can be used in the early stage of burn wound.Innovative:The molecular mechanism of asiaticoside in promoting burn wound healing and exerting anti-inflammatory effects was studied.The time when the burn wound began to be used was predicted.