Study On The Effect Of Indigo Naturalis Oil On The Model Of Psoriatic Lesions

ObjectiveTo establish methods for the quantification of indirubin content and preparation of indigo naturalis oil(psoriasis topical preparations).To discusse modeling period of the model of psoriatic lesions induced by propranolol hydrochloride.To evaluate the curative effect and security of the indigo naturalis oil to cure psoriatic lesions.Methods1.Indigo naturalis oil was prepared according to the ratio of indigo powder and camellia oil(1∶5,1∶10),mixture of camellia oil and olive oil(1∶5∶5),olive oil(1∶5,1∶10),by high temperature heating,extracting,filtrating.High performance liquid chromatography were used to determine the content of indirubin in all kinds of indigo naturalis oil.Column:Kromasil C18 column;mobile phase:methanol-10%aqueous N,N-dimethylformamide.The flow rate was1 mL/min;detection wavelength:292 nm.2.Propranolol hydrochloride liniment were used in guinea pigs(32),randomly divided into normal group(16)and model group(16).Each guinea pigs in model group were smeared with propranolol hydrochloride liniment on both sides of the auricle skin,three times a day,for five consecutive weeks.After two weeks,three weeks,four weeks,five weeks,both sides of the guinea pigs skin specimens of four normal guinea pigs and four model group were taken every week respectively,formaldehyde fixed,HE stained,light microscope observated,to evaluate the quality of the model of psoriatic lesions in guinea pigs induced by propranolol hydrochloride.3.Thirty-six healthy guinea pigs were randomly divided into a normal group(8)and a model group(28).The model of psoriatic lesions in guinea pigs induced by propranolol hydrochloride were chosen.The molding time was three weeks.After three weeks,four normal guinea pigs and four model guinea pigs were taken,and the pathological examination of bilateral auricle was performed to evaluate the quality of the model.Every eight ears on one side of the guinea pig of model group was set into one group.The group was selected as the test group,which external used indigo naturalis oil;the olive oil and camellia oil group was the matrix control group;the group was selected as the positive drug group,which external used calcipotriol liniment;all of the above groups used drug for four weeks and two times a day,the group control study was conducted with the normal group and the model group.Curative effect and adverse reaction were observated after four weeks.All the groups,bilateral auricle skin samples were taken,formaldehyde fixation,HE staining,light microscope observation,to evaluate the effect of each drug on psoriatic skin lesions model.Results1.In indirubin content determination,the linear range was 1.536-5.376μg/mL(r~2=0.9999).The average recovery was 99.4%and RSD was 2.2%.The indirubin extracted rates from indigo-camellia oil(1∶5),indigo-olive oil(1∶5),indigo-camellia oil(1∶10),indigo-olive oil(1∶10),indigo-olives-camellia oil(1∶5∶5)were 0.050%,0.086%,0.099%,0.144%,0.106%respectively.2.The typical psoriatic lesions of erythema appeared in two weeks.Pathologic examination showed hyperkeratosis of epidermis,thickening of acanthocyte layer,epithelial process with pestle,and lymphocyte infiltration in the superficial layer of dermis.The pathological changes of psoriatic lesions were more obvious in three weeks than that in two weeks,Baker’s histological score was higher than that of model in two weeks.During four to five weeks,the appearance of skin was hardened and atrophy.3.After model therapy(in the fourth week of the intervention),the psoriatic lesions of the calcipotriol group and the indigo naturalis oil were significantly alleviated,no obvious adverse reactions were observed,the Baker score was lower than that of the natural transfer model group,and difference was statistically significant(P<0.05).The Baker score of the matrix oil group was slightly lower than that of the natural transfer model group,but difference was not statistically significant(P>0.05).Conclusion1.Indigo naturalis oil can be made by mixing indigo naturalis with the olive oil or camellia oil in proportion,by high temperature heating,extracting,filtrating.The method of HPLC to determinate the content of indirubin which is the active ingredient of indigo naturalis oil is simple and feasible and can be used for the quality control of indigo naturalis oil.2.It is appropriate to select the model time for the model of psoriatic lesions for two to three weeks.3.Indigo naturalis oil(indigo naturalis olive oil,Indigo naturalis camellia oil)lesion is safe and effective to cure psoriatic lesions.The effect was similar to that of carpoolol.It deserves further research and application.