The Multiple Evaluation Of The Effect Of Shengji Ointment After Irradiation Sterilization

Objective: Investigate the effect of Shengji ointment after irradiation sterilization,in order to evaluate the feasibility of the irradiation sterilization process.Methods: 1.After modeling,rabbits were randomly divided into unirradiated and irradiated,treatment with the unirradiated or irradiated Shengji ointment.Wound healing was observed at 3d,6d,9d,12 d and 15 d.Take the blood and separate the drug loaded serum at same time.After 15 d,the wound healing skin was excised,make the pathological section and observe.In addition,the eligible subjects were randomly divided into two groups,dressing change,in the treatment of 0d and 9d,wound infection was analyzed by wound secretion.2.To establish the fingerprint of free amino acids in serum,and to analyze the changes of free amino acids in serum of different time points.3.The experiment of cell proliferation and apoptosis.Respectively with OD and apoptosis inhibition rate as the evaluation index,to analyze the effect of Shengji ointment after irradiation on the proliferation and apoptosis of human dermal fibroblasts.4.The above experimental data were analyzed by SPSS and Excel.Bivariate correlation analysis,multiple linear regression analysis and grey relational analysis were used to analyze the data.Established spectrum effect relationship between the free amino acid in serum and wound healing,in order to analyze the effect of irradiation on the therapeutic effect.Results: 1.The wound healing of the two groups were similar.Pathological examination showed that there were a lot of fibroblast growth in the two groups,and a large number of eosinophilic foci were found in the unirradiated group,and the number of capillaries was much more than that of irradiation group.There was no difference in the wound infection between the subjects（P=0.881,>0.05）.2.Establishment of serum free amino acid fingerprint by amino acid analyzer.Detection of 17 kinds of amino acid content,include: Tau,Thr,Ser,Glu,Gly,Ala,Val,Met,Ile,Leu,Tyr,Phe,Orn,Lys,His,Arg and Pro.Compared with blank serum,the maximum decrease of 15 free amino acids was more than 40% of two groups except for Tau and Glu,The decline range of the unradiation group ranged from 42.58% to 96.12%,while the irradiation group was from 41.95% to 90.64%,and the top 4 changes were Gly,Leu,Orn and Pro,and there was no difference between the two groups.From the analysis of the change of the time,the change process of the two groups was slightly different,but there was no difference in the overall trend.3.The two groups had obvious effects on promoting cell proliferation and inhibiting apoptosis.However,there are obvious differences in reaction time and intensity.4.To establish the spectrum effect relationship,comprehensive evaluation and analysis.Bivariate correlation analysis showed that Gly and Lys are closely related to cell proliferation（Pearson index was 0.725 and 0.695,P<0.05）;Orn was closely related to cell apoptosis inhibition rate（Pearson index was-0.653,P<0.05）.Multiple linear regression analysis: the cell proliferation rate is set to Y₁,the multiple regression equation:Y₁=-1.012-0.012X₃+0.004X₄+0.001X₆+0.016X₇+0.002X₁₃+0.000X₁₄-0.017 X₁₅-0.001X₁₆+0.000X₁₇（R=1.000）;apoptosis inhibition rate is set to Y₂,the multiple regression quation:Y₂=186.972+0.275X₃-0.032X₄-0.157X₆-0.429X₇-0.276X₁₃+0.275 X₁₄+0.262X₁₅-0.134X₁₆+0.113X₁₇（R=1.000）.Regression analysis showed that only 9 kinds of amino acids were related to the proliferation and apoptosis of fibroblasts,include: Ser,Glu,Ala,Val,Orn,Lys,His,Arg and Pro.Gray correlation analysis showed that the correlation between free amino acids and proliferation was sorted: Met>Lys>0.7>Orn>Ile>Phe>Pro>Val>His>Tyr>Glu>Leu>Ala>Tau>Ser>Thr>Gly>Arg>0.6;the correlation between free amino acids and the inhibition of apoptosis was sorted:Tau>Ser>Ile>Phe>His>Val>Tyr>Leu>Thr>Glu>Arg>Ala>Met>Lys>Pro>Orn =0.700>Gly>0.6.Three comprehensive statistical analysis results show that: Lys was positively correlated with proliferation of human dermal fibroblasts;Orn was significantly negatively correlated with apoptosis inhibition;Glu,Ala,Val and Arg were negatively correlated with apoptosis inhibition,Gly was unrelated to apoptosis inhibition.The changes of free amino acid content in the two groups were compared,there was no evidence of difference between irradiated and unirradiated Shengji ointment in the aspects of dermal fibroblast proliferation and apoptosis.Conclusion: It is feasible to sterilize Shengji ointment by irradiation.