| Part Ⅰ Theoretical Research Analysis of medication data for diabetic retinopathy treated by Chinese MedicineObjects:Traditional Chinese medicine has its unique advantages in the treatment of diabetic retinopathy(DR),and the therapeutic effect is remarkable,but there is a lack of systematic induction and analysis in medication.This study is based on the literatures of TCM treatment of DR in the past 10 years,and the prescriptions in the literatures are taken as the analysis object.The V3.0 software of traditional Chinese medicine inheritance calculation platform was used to analyze the drug property,taste,and medication rule of the prescription,and the association rule calculation formula was used to calculate the correlation between the two main components of "Astragalus" and "Angelica" in DR treatment.Methods:The literatures included in the 10-year database from 2010 to 2019 were selected.Key words related to diabetic retinopathy were selected for retrieval,and initial literature was obtained after preliminary screening.Using word and excel software to collect and organize the obtained data,and through the traditional Chinese medicine inheritance computing platform V3.0 to conduct frequency analysis on the properties,flavors,and meridians,the characteristics of high-frequency drugs for the treatment of DR can be preliminarily inferred.And preliminarily judged whether the two components in Danggui Buxue Decoction conform to the prescription characteristics related to DR treatment in the past 10 years.The correlation between Astragalus and Angelica was statistically calculated.Through the final data analysis,analyze whether the single-flavor drug components of Danggui Buxue Decoction meet the drug characteristics for DR treatment,calculate the improvement degree of the two single-flavor drugs astragalus and angelica,and determine whether the two prescriptions have clinical effects on DR treatment.research value.Results:Frequency of drug use: A total of 96 prescriptions were collected from 73 selected Chinese medicines,and a total of 148 Chinese medicines were obtained,with a total use frequency of 998 times.The top 10 drugs in use frequency were: rehmannia glutinosa,salvia miltiorrhiza,Astragalus membranaceus,Notoginseng,Chinese yam,wolfberry fruit,cortex moutan,cornus officinalis,Angelica sinensis and cooked Rehmannia glutinosa.The medicinal properties are mainly cold,followed by warm and flat,the taste of the medicine focuses on sweet,bitter and bitter,and the five meridians of liver,kidney,heart,lung and spleen are mainly.Calculation and analysis of correlation between Astragalus membranaceus and Angelica sinensis in Angelica Tonifying blood Soup: A total of 96 prescriptions were collected and 148 TCM ingredients were obtained,among which astragalus membranaceus appeared 37 times and Angelica sinensis 28 times,both of which appeared 14 times simultaneously in the same prescription.The confidence degree refers to the percentage of the occurrence times of the preceding and the latter items in the occurrence times of the preceding item: 14÷37,the improvement degree means the ratio of confidence and the percentage of the occurrence of the latter term in the total number of prescriptions is :(14÷37)/(28÷96)≈1.297.Conclusions:Through the final analysis of relevant literature,it was found that the single component of Danggui Buxue Decoction was in line with the drug characteristics of DR treatment,and the improvement degree of the formula characteristics of the two single drugs was greater than 1,and the formula of the two drugs was correlated with DR treatment and of high research value.Part Ⅱ Experimental study onExperiment 1 the effects of hyperglycemia on rat retinal microvascular endothelial cells at different stages in vitro were studied and the establishment time of hyperglycemia model was determinedObjects:To study the effects of hyperglycemia factors on different stages of rat retinal microvascular endothelial cells cultured in vitro and determine the establishment time of hyperglycemia model.Methods: The culture medium was divided into blank control group and high glucose and hypoxia group.The culture medium was added into each experimental group and cultured in a 37℃ CARBON dioxide incubator for 1d,2D and 4D,respectively,and the culture was terminated.MTT method was used to detect OD value,so as to detect cell proliferation and analyze and determine the time of successful establishment of the model.Results:On 1d,there was a statistically significant difference in OD values between the blank control group and the high glucose hypoxia group(P = 0.00),and OD values of the blank control group were greater than those of the high glucose hypoxia group,indicating that the high glucose hypoxia factor mainly inhibited cell proliferation at this time.At 2d,there was no statistically significant difference between the two groups(P = 0.12),indicating that the inhibitory effect of high glucose and hypoxia factors on the whole cell was weakened at this time.Compared with the normal group,it was found that the inhibitory effect on the whole cell tended to be zero at this time,neither promoting nor inhibiting cell proliferation.On day 4,the difference between the two groups was statistically significant(P = 0.00),and the OD value of the high glucose hypoxia group was significantly higher than that of the blank control group,suggesting that the high glucose hypoxia factor played a promoting role in cell enhancement at this time.Conclusions:Hyperglycemic hypoxia can inhibit the proliferation of rat retinal microvascular endothelial cells in vitro,but not in 2d,and promote the proliferation in 4D.The hyperglycemic hypoxia model was established in 4D.Experiment 2 the effect of Danggui Buxue Decoction on retinal microvascular endothelial cells of rats under hyperglycemia was studiedObjects:To study the effects of Danggui Buxue Decoction on the migration,proliferation and angiogenesis of retinal microvascular endothelial cells in rats under hyperglycemia.Methods:Select 3 generations in logarithmic phase of rat retinal microvascular endothelial cells,were randomly divided into blank control group,high oxygen hypoxia group,the sugar sugar + low-dose drug-containing serum group,high dose in sugar hypoxia +medicine serum group,high sugar hypoxia + high dose drug-containing serum 5 group,each group shall have three parallel hole,join the experiment culture of each group,Endothelial cell migration experiment,endothelial cell proliferation experiment and endothelial cell angiogenesis experiment were carried out.After being placed in a 37℃CARBON dioxide incubator for 4 days,the culture was terminated and tested.The experiment was repeated for 3 times.Results:On the 4th day,the number of cell migration,proliferation and angiogenesis increased in order from the blank control group,the high,medium and low concentration drug-containing serum groups,and the high glucose hypoxia group,suggesting that the high glucose hypoxia factor has an adverse effect on the retinal microvessels of rats.Endothelial cells play a positive role in promoting.After the application of Danggui Buxue decoction containing serum,the positive promoting effect of high glucose and hypoxia factors on cells is improved,and with the increase of drug concentration,the improvement effect is gradually enhanced.At low concentrations,this drug has limited effect,and the difference with the high glucose and hypoxia group is not statistically significant,but as the concentration increases,the effect becomes more and more significant,and the difference between different concentration groups is statistically significant.When compared with the blank control group,the best curative effect was achieved,and there was no statistically significant difference between the two.Conclusions:The experiments of cell migration,endothelial cell proliferation and endothelial cell angiogenesis showed that high glucose hypoxia mainly promoted retinal microvascular endothelial cells in 4d,and danggui Buxue Decoction containing drug serum could also resist this promoting effect,which was positively correlated with the concentration.Experiment 3 Study on the protective mechanism of Danggui Buxue Decoction on rat retinal microvascular endothelial cells under hyperglycemiaObjects:To study the mechanism of Danggui Buxue Decoction on rat retinal microvascular endothelial cells under hyperglycemia through il-1 β→COX-2/PI3K/AKT→VEGF pathway.Methods:Select 3 generations in logarithmic phase of rat retinal microvascular endothelial cells,were randomly divided into blank control group,high oxygen hypoxia group,the sugar sugar + low-dose drug-containing serum group,high dose in sugar hypoxia +medicine serum group,high sugar hypoxia + high dose drug-containing serum 5 group,each group shall have three parallel hole,join the experiment culture of each group,After being placed in a 37℃ carbon dioxide incubator for 4 days,the culture was terminated,and the protein and m RNA contents of PI3 K,Akt,IL-1β,VEGF and COX-2 in each group were detected by Western blot and fluorescence quantitative PCR,respectively.Results: Western blot and fluorescence quantitative PCR showed that the protein and m RNA of retinal microvascular endothelial cells were mainly promoted by high glucose hypoxia at 4d,while Angelica Buxue Decoction could resist this promoting effect and was positively correlated with the concentration.Conclusions: Dangguibuxuetang drug serum may reduce the gene expression levels of IL-1β,COX-2,PI3 K,AKT,VEGF and IL-1β,COX-2,p-PI3 K,IL-1β,COX-2,p-PI3 K,The expression levels of p-Akt and VEGF protein play a role in inhibiting the migration,proliferation and angiogenesis of retinal microvascular endothelial cells,which proves that Danggui Buxue Decoction can inhibit retinal neovascularization through the IL-1β→COX-2/PI3K/AKT→VEGF pathway To achieve the effect of preventing and treating PDR. |
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