In Vitro And In Vivo Experiments Of A Novel Photosensitizer Ye Luguang-1 Mediated Photodynamic Therapy For Killing HP

Objective:1、To clarify the inhibitory effect of photosensitizer-mediated photodynamic therapy on Helicobacter pylori.2、Determining the best effect parameters of Ye Luguang No.1 mediated photodynamic therapy for killing HP;The lowest drug concentration for killing HP;the relationship between different concentrations of photosensitizer drug concentration against Helicobacter pylori activity;3、Constructing HP infectious gastritis pig model,the next step of Ye Luguang 1 mediated photodynamic therapy for HP infection in vivo Research is the foundation.4、To clarify the therapeutic effect of PDT on HP infectious gastritis.Methods:The prepared sterile photosensitizer-containing paper,the positive control clarithromycin tablet and the blank paper were grouped and placed on the surface of the blood plate coated with 100 ul HP,Using a P652 II type semiconductor laser therapeutic apparatus,adjust the appropriate spot diameter to irradiate the surface of the HP blood plate,the power is 600 m W,the power density is 7.64 m W/cm2,and the irradiation is20 min,Through the diameter of the zone of inhibition,the photosensitizer Ye Qingguang I（HCE6）-mediated photodynamic therapy inhibits Helicobacter pylori.2、Prepare the HCE6 photosensitizer solution,and dilute the concentration of 100 ul of HCE6 photosensitizer in 1-12 to D rows 1-12 of A row in 96-well plate to 200 ug/ml,100 ug/ml,50 ug/ml,25ug/ml,respectively,using a continuous 2-fold dilution method.1-12 wells of E line is 100 ul blank control group without HCE6,and 100 ul of the configured HP bacterial suspension is respectively added into the above hole;Using P652 II type semiconductor laser therapeutic apparatus,adjust the appropriate spot diameter,power 600 m W,power density is 7.64 m W/cm2,irradiate 96-well plate,irradiation time 1-3column for 0min,4-6 column for 10 min,7-9 column for 20 min,10-12 column for 30 min,after the operation is completed,the 96-well plate is transferred to a37°C micro-aerobic environment for 48 h.After 48 h,different concentrations of photosensitizer and different irradiation time solutions were diluted to the corresponding multiples,and 100 ul coated blood culture dishes were aspirated.After 72 h,the number of HP colonies on the surface of the blood culture dish was counted.Statistical analysis was carried out to determine the optimal effect parameters.3、The 10 normal piglets purchased were randomly divided into two groups: blank control group,dexamethasone + HP group 4,dexamethasone + ranitidine + HP group 4,to construct HP colonization infected pig model;Blood HP antibody,gastroscopic observation and clamping of mucosal tissue for rapid urease test,pathological examination of HE staining,and mucosal HP isolation and culture confirmed that HP had colonized the gastric mucosa of infected piglets.Photodynamic therapy was performed after the HCE6 photosensitizer was sprayed through the gastroscope.The morphology of gastric mucosa before and after treatment was visually observed under gastroscopy.The therapeutic effect of HCE6 on HP customization and infection-associated gastritis was confirmed by blood HP antibody detection,rapid urease test and HE staining pathological examination.Results:1,Group A（simple photosensitizer）diameter= 6.10 + 0.02 mm;Group B（simple light）diameter = 6.51 + 0.07 mm;Group C（photodynamic therapy）diameter = 21.07 ± 0.19 mm;Group D（Clarithromycin）diameter = 24.80 ± 0.21 mm;2,when PDT irradiation power P = 600 m W,HCE6 concentration = 200 ug / ml,irradiation t1 = 20 min,sterilization rate = 96.5%;t2 = 30 min,sterilization rate = 96.6%;when PDT irradiation power P = 600 m W,irradiation time t = 20 min,concentration = 100ug/ ml,sterilization rate = 90.5%,concentration At 200 ug / ml,the sterilization rate was 96.5%.3.In the experimental group（dexamethasone + ranitidine + HP group）,3 out of 4 piglets successfully constructed a model of HP colonization infection in normal piglets.4,HCE6-PDT treatment of HP infected piglets,improved gastritis,HP gastric mucosa colonization disappeared Conclusion: 1.The photodynamic therapy mediated by the novel photosensitizer Yelvguang I has a strong inhibitory effect on HP in vitro.The optimal effect parameters are p=600m W,t=20min,and the photosensitizer concentration is 200ug/ml.2.Helicobacter pylori SS1 strain can colonize and infect common piglets.HCE6-mediated photodynamic therapy is a treatment method for effectively killing Helicobacter pylori in piglets and treating HP-related gastritis.