The Effect Of Philippine Bull Leeches Extract On Melanoma Cells And Its Antioxidant Activity And Safety Evaluation

Objective:To study the effect and antioxidant activity and safety evaluation of the extracts of Poecilobdella manillensis on Melanoma cells,in order to provide a new idea for the study of The effect of the white anti-decay of Poecilobdella manillensis,and to lay a foundation for the development of new drugs and cosme-tics.Methods:1.In vitro,the effects of the extracts of Poecilobde-ll manillensis on the proliferation of mouse melanoma cells,the effect of tyrosinase activity in cells and the inhibitory effect on melanin formation were studied by the method of culturing mouse melanoma cells.2.The skin of nude mice was irradiated by ultraviolet light,and the skin pigmentation model was established,and the method of skin smear samples in nude mice was observed,the skin tissue was taken after administration,the paraffin section was fixed,and the activity of sod,MDA and TYR in plasma were detected.3.The removal of hydroxyl radical,superoxide anion free radic-als and DPPH free radicals by the extracts of Poecilobdella manill-ensis was detected by spectrophotometry,so as to evaluate the abil-ity of the extracts of the Poecilobdella manillensis to remove free radicals.4.The acute toxicity of intact and damaged skin was studied by using the method of dorsal hair-feeding in rats.Observe the ani-mal's weight,skin,changes in hair,eyes and mucous membranes, respiration,circulation,central nervous system,and limb movements,and the use of both sides of the rat back to the hair treatment method,the drug single and many times outside the normal or damaged hair removal area of the back of the rat,observe its skin irritation reaction to animals,the skin allergy test was carried out by using the method of hair treatment on both sides of guinea pig,the allergic reaction of local medication was observed,and the average value and sensitization rate of the reaction were calculated.Results:1.In vitro cell experiments,compared with different concentrations at the same time,the inhibition rate of Poecilobdella manillensis extact and arbutin on mouse melanoma cells and inhibi-tion rate of tyrosinase activity with the increase of drugs concentra-tion increase;the same concentration at different times for compari-son,the inhibition of Poecilobdella manillensis extract and arbutin on mouse melanoma cells and inhibition rate of tyrosinase activity increased with time;and Poecilobdella manillensis extract inhibits mouse melanoma cells and inhibits tyrosinase activity more strongly than arbutin.The inhibitory rate of melanin content in mouse mel-anoma cells in a dose dependent manner was determined by the extract of burdock burdock and arbutin,and the inhibitory effect ofPoecilobdella manillensis extract on melanin formation in mous e melanoma cells wascorrelated with the arbutin content.Suppressi on ability is not muchdifference.The inhibitory rate of Poecilobdell a manillensis extractand arbutin on HUVEC cells increased with ti me.2.In vivo experiments,compared with the blank group,compared with the model group,the number of melanocytes in the skin of high-dose nude mice was significantly reduced?P<0.05?,and the content of MDA in plasma was significantly reduced?P<0.05?.The SOD content in plasma increased?P<0.05?,and the activity of TYR in plasma decreased significantly?P<0.05?,indicating that the extract of burdock has a good therapeutic effect on skin pigm-entation.3.In the in vitro antioxidant experiment,the DPPH radical scavenging rate of Poecilobdella manillensis extract increased with the increase of drug concentration,and the IC₅₀ of half concentrat-ion was 0.45 mg/mL,which was higher than the IC₅₀ of vitamin C?0.07 mg/mL?.Visible,Poecilobdella manillensis extract has a stro-ng DPPH scavenging ability;Poecilobdella manillensis extract on hydroxylradical scavenging rate increases with the increase of drug concentration,half inhibition concentration IC₅₀ is 0.49mg/mL,higher than vitamin C The IC₅₀?0.07 mg/mL?showed that the extract of burdock has astrong hydroxyl radical scavenging activity;the scave-nging rate of superoxide anion by extracts of burdock increased with the increase of drug concentration,and the flavonoid extract was The half-inhibitory concentration IC₅₀ was 0.43 mg/mL,which was highethan the IC₅₀?0.08 mg/mL?of vitamin C.It can be seen that the Poecilobdella manillensis extract has strong superoxide anionradical scavenging ability.4.In the acute toxicity test,the skin was exposed to ephedrine extract for a short period of time.There was no abnormality in respiratory status,mental status,appearance,and behavior.There was no abnormal reaction in the skin of the medication site,and there was no significant change in body weight?P>0.05?;In the irritant experiment,no edema or erythema occurred in the skin of the back of each group of rats after single or multiple doses.The average response score was 0,showing no skin irritation,indicating that the Poecilobdella manillensis extract was not irritating to rat skin;in the skin allergy test,guinea pigs in the high,middle,and low doses of Poecilobdella manillensis had no allergic skin erythe-ma and edema,nor had asthma,standing instability,or shock,etc.The allergic reaction showed that the Poecilobdella manillensis extract was not allergic to transdermal administration.Conclusion:1.The effects of the extracts of the Poecilobdella manillensis on the proliferation,activity of tyrosinase and formation of melanin in mouse melanoma cells were inhibited.2.The Poeci-lobdella manillensis extracts have a good therapeut ic effect on the skin pigmentation of nude mice.3.The ability of DPPH free radi-cals,hydroxyl radicals and superoxide anion free radicals to be removed from the extracts of Poecilobdella manillensis showed that the extracts of the Poecilobd-ella manillensis had antioxidant effects.4.The Poecilobdella manillensis extracts are safe without toxici-ty,irritationand allergic property by transdermal delivery.