Efficacy Evaluation Of Qufeng Fuzheng Solution In The Treatment Of Peripheral Facial Paralysis In Rats Study On Toxicity Tests Of Single And Repeated Administration

Objective:Peripheral facial paralysis rat models were developed by crushing injury,aimed at evaluating the therapeutic effect of Qufeng Fuzheng solution on peripheral facial paralysis treatment.Meanwhile,the toxic reactions were investigated in terms of property,degree,dose-effect relationship,time-effectiveness relationship and reversibility through single-and repeat-dose toxicity studies for the toxic target organs or tissues determination and ultimate safety evaluation of Qufeng Fuzheng solution.Methods:1.In the efficacy evaluation part sixty rats were randomly divided into blank group,model group,Qufeng Fuzheng solution low-dose group(0.03 m L/kg),medium-dose group(0.06 m L/kg),high-dose group(0.12 m L/kg),and prednisone acetate group(4mg/kg),with 10 rats in each group.The experimental group was administrated intranasally and the prednisone acetate group was administrated by oral gavage for successive 21 days.Blink reflex,tentacle movement,and tentacle direction were evaluated by three-point scoring method,and the effect of Qufeng Fuzheng solution on the behavior of rats were observed.2.In the single administration toxicity tests 10 SD rats were distributed to each group with male and female equally divided.The intranasally administration group was administrated at 3 m L/kg/d,and the oral gavage administration group was administrated at 2 m L/kg/d ～ 5 m L/kg/d.Observation was performed for 4 h after administration and once a day during the recovery period.Body weight was measured before administration and on the 1st,2nd,3rd,7th,14 th day of recovery period,and food intake was measured before administration and on the 1st,2nd,13 th day of recovery period.Blood was collected before administrationat and at 2 min,10 min,30 min,1 h,2 h and 4 h after administration in the intranasally administration group.And blood was collected before administrationat and at 30 min,1 h,2 h,4 h,6 h,8 h and 12 h after administration in the oral gavage group.The contents of benzoylaconitine,and benzoylhyponitine in plasma of rats were determined by LC-MS/MS method.Gross anatomy was performed on the 15 th day of recovery period.3.In the repeated administration toxicity tests 30 SD rats were distributed to each group with male and female equally divided.The test was divided into low dose group(0.1 m L/kg/d),medium dose group(0.33 m L/kg/d),high dose group(1 m L/kg/d),nasal administration group(1 m L/kg/d)and blank control group.The drug was administered once a day,and withdrawal for 28 days after continuous administration.The general state of the rats was observed daily,and body weight and food intake were measured regularly.Ophthalmology,hematology,blood biochemistry,urinology and histopathology were performed at the end of the administration period and recovery period.In order to investigate the relationship between drug dose and drug exposure,blood was collected before administration and at 2 min,10 min,30 min,1 h,2 h,4 h in intranasally administration group,and before administration and at 30 min,1 h,2 h,4 h,6 h,8 h,12 h in oral gavage administration group.The active components of Qufeng Fuzheng solution in plasma was determined by LC-MS/MS method.Results:1.After 21 days of treatment,blink reflex,tentacle movement and direction function of facial paralysis rats gradually recovered.Total score of treatment groups was higher than that of model group,and efficacy score of Qufeng Fuzheng solution low-dose group(0.03 m L/kg)was lower than that of prednisone acetate group(P < 0.05).2.The weight gain of 4 m L/kg oral gavage administration group was slightly slower than that of other groups.The intake of oral gavage administration group decreased with the increase of administration volume.The results of gross anatomy showed that there was no obvious abnormality in the external features of the rat body,and no obvious abnormality in the color,size,position and texture of the viscera.The LD50 was 3.323 m L/kg(95% confidence limit: 2.236 ～ 4.034 m L/kg),which was equivalent to the LD50 of raw medicinal materials 4.15 g/kg(95% confidence limit: 2.795-5.043 g/kg).The results of toxic dynamics showed that the exposure levels of the three components in plasma were lower in intranasally group than those in the oral gavage administration,and the plasma concentration was lower than the lower limit of quantification 1 hour after administration.3.The rats in oral gavage administration groups and intranasally administration group were repeatedly given Qufeng Fuzheng solution for 28 consecutive days.All rats were in normal state,and no toxic reactions related to Qufeng Fuzheng solution were found in body weight,food intake,urinology,hematology,hematochemistry,ophthalmology and histopathology.After one hour of intranasally administration,the blood concentration of components were all lower than the lower limit of quantitation of0.5 ng/m L.In vivo exposure was positively correlated with dose at 0.1 m L/kg,0.33 m L/kg and 1 m L/kg oral gavage doses.However,only 1 m L/kg of oral gavage administration could detect all three compounds from 0.5 h to 4 h after administration.No toxic reaction was observed after repeated administration of Qufeng Fuzheng solution 1m L/kg/d at 28 days.Conclusion and significance:Qufeng Fuzheng solution showed an excellent therapeutic effect on peripheral facial paralysis and could effectively promote the recovery of symptoms after facial paralysis.At the same time,Qufeng Fuzheng solution has good safety in single-and repeat-dose at clinical dosage.This study provides a theoretical basis for the development of Qufeng Fuzheng solution in the treatment of peripheral facial paralysis.