Dose-Effect Relationship Study Of Classical Prescription-Gegen Qinlian Decoction And Maxing Shigan Decoction

The definition of the dose-effect relationship of drug is the intensity changes of drug effect with the dose in certain dosage range, when the strength of effect reached the maximum, the dose increased but effect did not change. The formula dose is one of the key scientific issues and relates to clinical therapeutic effect. In the theoretical system of traditional Chinese medicine, the knowledge about the dose-effect relationship is sporadic and different to build system science theory of the clinical prescription dose. The theory of formula is important to guide clinical medication and improve the clinical level.According to the TCM holistic theory, Gegen Qinlian decoction (GQD) and Maxing Shigan decoction (MSD) were used as the study models. The dose-effect relationship of GOD and MSD on related animal models was investigated on the disease models include type2diabetes, ulcerative colitis, fever, cough and asthma.The experiment is divided into five parts. The corresponding dose-response characteristics of GQD and MSD were investigated in the models of different disease.1> Study on the dose-effect relationship of Gengen Qinlian Decoction in type2diabetes rats[Objective]To investigate the effects of Gengen Qinlian Decoction (GQD) in type2diabetes rats, body weight, organ index, blood glucose levels, insulin levels, insulin resistance (IR) index, glycosylated hemoglobin(GHb) levels, glycosylated serum protein (GSP) concentration, serum creatinine, serum urea nitrogen and other indicators of impact were investigated to obtain relative dose-effect relationship.[Methods]The rat model of type2diabetes was built. After the male SD rats (120-150g) were adaptively feed, they were feed by high fat food for4weeks to obtain the rat model of type2diabetes whereas the normal control rats were normal feed. Body weight of the rats from high fat treatment is more than120%body weight of the rats from normal control or physique quantum BMI (body weight/length of body square) of rats have statistics difference normally comparing the normal group. The rats were injected with30mg/kg streptozotocin (STZ) citric acid buffer solution by tail intravenous injection to induce the rat model of diabetes, the normal control group were injected the same level dosage the citrate buffer solution. After rats were treated for72hours and one week, empty stomach blood sugar value of the rats tail point were determined, Both of blood sugars level are more than11.1mmol/L so that the model of type2diabetes rats were built successfully. Experimental groups were divided as normal control group, type2diabetes mellitus model control group, positive control group (metformin hydrochloride200mg/kg) and GQD dose group (9of the dose change of the whole party were1.65、4.95、8.25、11.55、14.85、18.15、21.45、24.75、28.05g Pieces/kg)(n=15). Efficacy indicators were observed such as weight, organ indices (liver, kidney, spleen), blood glucose, insulin levels, glycated hemoglobin levels, and glycosylated serum protein concentration, serum creatinine, blood urea nitrogen etc.[Results]After oral administration for10weeks, serum glucose, insulin levels, and glycosylated serum protein concentration, glycosylated hemoglobin level, serum creatinine, blood urea nitrogen of type2diabetic rat were changed by GQD in the dose-effect relationship, and other indicators were demonstrated for dose-effect relationship. Based on level of blood glucose, the dose-effect relationship parameters of GQD on diabetic model rats were as follows:dose range:[D]0.8～[D]0.2=27.01～1.09g/kg;[D]0.8/[D]0.2=7; median dose:[D]0.5=3.81g/kg; dose threshold:0.2=1.09g/kg. Certain dose range with GQD was demonstrated that dose increases, drug efficacy enhancement.[Conclusion]Fine dose-effect relationship was confirmed by GQD treatment of type2diabetes. 2、The study of dose-effect relationship of Gegenqinlian decoction for the treatment of ulcerative colitis rats[Objective] To study the dose-effect relationship for the treatment of UC rats caused by dose change of Gegenqinlian decoction (GQD), and explore the dose parameters.[Methods]10SD rats was selected as control group,110SD rats were used to make ulcerative colitis (UC) model by the mixed liquid of5%TNBS an ethanol, then divided into11groups randomly, the MPO, MDA, NO, SOD content in plasma were detected, the SPSS and GraphPad software were used to fit dose-response curve, evaluate the dose-effect relationship, and calculate dose parameters.[Results] Take MPO content as the indicator, the fitting equation model similarity is high, and dose-response curves showed a reverse "S" shape, the calculated dose threshold parameters are as follows:[D]20～[D]80-(15.39～17.11)g/kg, median dose:[D]50=16.25g/kg, and the threshold dose:[D]20=15.39g/kg.[Conclusion] The dose-response curves for the GQD treatment rendered reverse "S" shape, suggesting that the effective range of treatment is the medium dose, roughly in (15.39～17.11)g/kg. 3、Study on the Dose-effect Relationship of Maxing Shigan Decoction (MSD) on the Treatment of Fever[Objective] To study the effect of Maxing Shigan decoction (MSD) with different dose on the treatment of the fever, and explore the dose-effect relationship to provide the scientific basis for the clinical application.[Methods]:The SD rats were grouped into11groups randomly:normal group, modeling group, and MSD dose-1group to dose-9group, set the fever model by inject lippolysaccharide (LPS) through the tail vein of rats. The temperatures of rats a time every0.5hour were recorded, and TRI6.0was calculated. After6.0hours treatment, after modeling, the concentration of PGE2in the plasma of the rats was detected. Metabolomics was used to observe the effects of MSD by the overall state of endogenous substances[Results]:The optimum LPS dose of fever model was20μg/kg. Compared with modeling group, dose-1group to dose-9group reduced temperature significantly (P <0.05,or P<0.01); the thermal response index (TRI) were dramatically decreased except dose-3group and dose-6group (P<0.05, or P<0.01); the level of PGE2in the plasma were lower than the modeling group(P<0.05, or P<0.01), and were not different from the control group (P>0.05).Three clinical relevant doses have effect on treating the fever of modeling rats. Low-dose group (LDG) could were reduced the temperature more than middle-dose group (MDG) and high-dose group (HDG), but the difference between three dosages was not significant. The TRI6.0increased with the dosage, but the antipyretic effect was decreased; and the PGE2level in the plasma of MDG was lower than LDG and MDG.From the results of metabonomics, there is an obvious dose-effect characteristic by the overall effect of endogenous substance state in fever model rats on the treatment of MSD. The dose-effect relationship parameters of MSD on fever model rats were as follows:dose range:[D]0.8-[D]0.2=4.0935g/kg-4.1543g/kg;[D]0.8/[D]0.2=1.015; median dose:[D]0.5=4.124g/kg; dose threshold:0.2=4.0935g/kg;[Conclusion]:MSD have antipyretic effect on rats which are set by LPS, but the mechanism of MSD treatment may not mainly by decreasing the synthesis of PGE2. The clinical dosages also have antipyretic effect on the rat models, and the dose-effect relationship can provide a reference to the clinical application. 4, The dose-effect relationship investigation of Maxing Shigan Decoction (MSD) on cough model of rats[Objective] To investigate the dose-effect relationship characteristics of MSD on the cough model of rats.[Methods] The guinea pigs were chosen as trail animal and modeled to cough by the citric acid solution with0.4mol/L. the solution was atomizated by ultrasonic atomizer with atomization rate of15L/min for1min. After that, the cough frequency was recorded for5min, the guinea pig with cough frequency beyond10times was selected as a model animal. In this way,120guinea pigs were selected as model animal and were randomly divided into12groups, named respectively by model group, positive group and dose1to dose9group. What is more, control group was settled. As a result, there are a total of12groups,10guinea pigs in1group.Control group were gavaged with NS, positive were gavaged with codeine phosphate (8.5mg/kg), dose1to dose9groups were gavaged with MSD for continuous administration for15days according to10ml/kg. After oral administration, cough frequency were recorded every10min in one hour, latent period with a stopwatch, variations of the amplitude of synchronous were recorded by biological signal acquisition system. At the same time, gas cough sound were with the sound amplifier, bronchial and lung tissue, blood centrifuged serum tested CAT, H2O2, MDA, SOD and other biochemical indexes.[Results] MSD has an obvious therapeutic effect in the clinical low and medium dose. Moreover, result showed that the therapeutic effect increased from1.44g/kg to4.286g/kg but high dose is invalid.[Conclusion] MSD has the effect on the treatment of cough. In a certain range, dose-effect curve is more twists and turns, the therapeutic effect increased with the gavaged dose. 5、Study of the Dose-effect Relationship of Maxing Shigan Decoction (MSD) on the treatment of Antiasthmatic[Objective] To study the antiasthmatic efficacy of MSD which exists as different doses of the whole prescription and different ratio of monarch drug and investigate the dose-effect relationship, provide an objective scientific basis for its reasonable amount of Clinical Practice.[Methods] Guinea pig experimental animal model was built by spraying volumetric mixture of histamine phosphate and acetylcholine. Both the whole prescription group and different drug ratio group are divided into12groups including the blank group, the model group, the positive control group and9MSD groups with different concentrations. Among every group, except the blank group and the model group lavaging physiological saline, we measure the asthma latency and asthma duration one hour after the last time lavaging with different concentrations of MSD. Results from relative groups were analyzed, and the chart of dose effect relationship was made. ELISA (enzyme linked immunosorbent assay) was applied for measuring the concentrations of HIS、LTB4、IL-4、IFN-y from the serum of anaesthetic guinea pigs, statistical analysis was performed by the software SPSS. The histopathologic examination of bronchial and lung tissue was performed. The amplitude change rate of asthma waves and calm waves were calculated and the criteria of asthma was set. Changes in tension of tracheal smooth muscle by the treatment with different concentrations of MSD were detected by isolated organ tester.[Results] Compared with the blank group, rapid onset asthma model group and late onset asthma model, it appear extremely significant difference according to the result of the three guinea pig asthma models (P<0.01), the repeated sensitizing late-onset asthma model group has no significant difference with the blank group (P>0.05)We took the time when the tic wave (changing rate≥1000%) appeared for the first time as standard so that to determining the incubation period of asthma and the time when the tic wave(changing rate≥100%) appeared for the last time as standard to assess the asthma duration.The medium dose group (3.24g/kg) and the fourth ratio dose group had good anti-asthmatic effects on the asthmatic model of guinea pig, which had extremely significant difference between the model group (P<0.01),and no significant difference between the positive control group (P>0.05)From the results of metabonomics, there is an obvious dose-effect characteristic by the overall effect of endogenous substance state in asthma model rats with the treatment of MSD. The parameter of dose-effect relationship of MSD on asthma model rats were as follows:dose range:[D]0.8-[D]0.2=1.3311g/kg～1.3415g/kg;[D]0.8/[D]0.2=1.008; median dose:[D]0.5=1.336g/kg; dose threshold [D]0.2=1.3311g/kg.[Conclusion] MSD has positive effects on the asthmatic guinea pig models which are sensitized by histamine phosphate and acetylcholine. The medium dose or small dose may be the optimal dose for clinical treatment of asthma. The dose-effect relationship of MSD may be helpful to clinical medication.