| Purpose:The effect of different time of Nepeta schisandrae on the content of pulegone in Nepeta decoction was determined by HPLC;the animal models of colds were replicated by the methods of heating,acute inflammation,induced pain,and coughing to observe Nepeta The effect of different time after the different on the antipyretic,anti-inflammatory,analgesic,antitussive and immune effects of Nepeta decoction.To investigate the best decoction time of Nepeta in Nepeta decoction,provide experimental basis for the clinical application of Nepeta special decoction method.Method:The effect of different boiling time of Nepeta on the menthol content in Nepeta decoction was determined by HPLC.The chromatographic conditions were:Inertsil ODS-3column(250×4.6 mm,5μm),mobile phase:methanol-water(80:20)Column temperature30°C,flow rate 1mL/min,detection wavelength 252nm.15%yeast suspension heating method,egg white induced acute inflammation method,hot plate induced pain method,and25%ammonia water-induced cough method were used to replicate cold-related animal models.Body temperature,toe swelling rate,pain threshold,and cold-related model animal were observed.Cough latency,number of coughs,spleen coefficient,thymus index,changes in lymphocyte subsets(CD4,CD8)in the spleen,and levels of immune factors(IL-1,IL-6)in the serum.Result:HPLC method showed a good linear relationship(r~2=0.9995)between peak volume and injection volume of 1~15μL(0.143~2.145μg)of the control solution of pulegone,and the decoction of No.1(after Nepeta decoction)In the next 2.5 min group)the content of pulegone in the test solution was 0.738 mg/g;No.2 decoction(in the next 5 min group after the Nepeta decoction)The content of pulegone in the test solution was 0.896 mg/g;Decoction(after 7.5 min group of Nepeta decoction)The content of pulegone in the test solution was1.040 mg/g;Decoction No.4(10 min after Nepeta decoction)was used for the determination of pulegone in the test solution.0.827mg/g;Decoction No.5(12.5min after the Nepeta decoction)The content of pulegone in the test solution is 0.713mg/g;Decoction No.6(15min after the Nepeta decoction)The pulegone content in the solution was 0.677 mg/g.The body temperature test showed that compared with the control group,the body temperature was significantly increased at 1 h,2.5 h,3 h,4 h,5 h,6 h,7 h,and 8 h after injection of 15%yeast suspension in the model group(P<0.05 or P<0.001).Compared with the model group,positive control group 1(aspirin enteric-coated tablets),positive control group 2(influenza Shufeng capsules),Nepeta 5min,7.5min,and 10min groups were injected 15%after yeast suspension 1h At 2.5 h,3 h,4 h,5 h,6 h,7 h,and 8 h,the body temperature decreased significantly(P<0.05 or P<0.01 or P<0.001);the 2.5 min group of Nepeta rose after injecting15%yeast suspension At 1 h,4 h,5 h,6 h,7 h,and 8 h,body temperature decreased significantly(P<0.05 or P<0.01 or P<0.001).At 12.5 min and 15 min after Schizonepeta,the body temperature was significantly lower at 4 h,5 h,6 h,7 h and 8 h after injection of 15%yeast suspension(P<0.01 or P<0.001).Compared with the control group,the toe swelling rate of the model group was significantly higher than that of the blank control group at 0.5h,1h,2h,3h,and 4h after the injection of egg white(P<0.001);compared with the model group,the positive control Group 1(aspirin enteric-coated tablets),positive control group 2(influenza cold-wind capsule),and 2.5 min,5 min,7.5 min,and 10 min groups of Nepeta,after the injection of egg white 0.5h,1h,2h,3h,toes The swelling rate was significantly decreased(P<0.05 or P<0.01 or P<0.001);the Swelling rate of the toes was significantly decreased at0.5 h,2 h,3 h,and 4 h after the injection of egg white in the 12.5 min group of Nepeta(P<0.01 or P<0.001);After 15 min of Nepeta group,0.5h and 2h after the injection of egg white,the swelling rate of the toe was significantly decreased(P<0.001).Pain threshold test,compared with the model group,the positive control group 1(aspirin enteric-coated tablets),positive control group 2(influenza cold wind capsule),Nepeta later 2.5min,5min,7.5min group pain threshold increased significantly(P<0.001).Cough latency and number of coughs were compared with those of the model group.The positive control group 1(Pentamicin citrate tablets),the positive control group 2(influenza Shufeng Capsule),and Nepeta later 2.5min,5 min,7.5 min,The latency of cough was significantly higher in the 10 min group(P<0.001);the positive control group 1(citronoxetine citrate),the positive control group 2(influenza Shufeng Capsule),and 2.5 min,5 min,and 7.5 min after the Nepeta The number of coughs in the 10min,12.5min and 15min groups was significantly lower(P<0.01 or P<0.001).Spleen and thymus index test,compared with the model group,the spleen coefficient of the positive control group 1(citronovolate citrate tablets),positive control group 2(influenza Shufeng capsule),7.5min,10min group after nepeta rose significantly High(P<0.01 or P<0.001);positive control group 1(Pentoxifene Citrate Tablets),positive control group 2(influenza Shufeng Capsule),and thymus index after 7.5 min,10 min,and 15 min groups of Nepeta Significantly increased(P<0.05 or P<0.001).Flow cytometry test,compared with the model group,positive control group 1(citronoxetine citrate),positive control group 2(influenza cold wind capsule),Nepeta later 2.5min,5min,7.5min,10min The level of CD4 in the 12.5 min group was significantly higher(P<0.05 or P<0.001);the positive control group 1(citronoxetine citrate),the positive control group 2(cold Shufeng capsule),and the next 5 min.The levels of CD8 in the 7.5 min and 10 min groups increased significantly(P<0.001).Enzyme-linked immunosorbent assay(ELISA)test,compared with the model group,positive control group 1(citronovolate citrate tablets),positive control group 2(influenza cold wind capsule),Nepeta later 2.5min,5min,7.5 The levels of IL-1 were significantly increased in the min,10min,and 12.5min groups(P<0.05 or P<0.01 or P<0.001);Positive control group 1(Crotecorate citrate tablets)and Positive control group 2(cold After Feng Capsule,IL-6 levels in the 2.5 min,5 min,7.5 min,10 min,and 12.5 min groups increased significantly(P<0.05or P<0.01 or P<0.001).Conclusion:During the decocting process,the highest pulegone content in Nepeta decoction was 7.5 min after Nepeta paniculata,which significantly improved body temperature,toe swelling,pain threshold,cough latency,cough frequency and immune function in cold model animals.effect.The use of special decoction method(ie,"after the next")in the Nepeta decoction can achieve enhanced pharmacological effects,reduce toxic side effects,and improve clinical efficacy. |
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