| This research was a part of the item supported by fund for key research project of national natural science foundation of China, Studies on compatibility principles of Mahuangtang(MHT).Objective:To study the diaphoretic function of MHT and its mechanisms in animals,and to illuminate compatible theories of MHT.Methods: The sample position was confirmed by comparing axillary with plantar sweat gland.Duct diameter of axillary sweat gland was taken as an index to estimate sweating quantity by tectology. The diaphoretic function of different compatibility of MHT was observed at 30 minute after administration,The sweating volume,incidence of vacuole and duct diameter of axillary sweat gland were taken indexes to estimate sweating quantity by tectology.Effects of MHT on anal temperature ,auricle microcirculation,adrenergic receptor,M receptor in mice were observed too.Left sciatic nerve was severed by operation in rats.The diaphoretic function on left thenar sweat gland was observed at 30 minute after administration of MHT by tectology .Results: 1.The methods of diaphoresis experiment:(1)The sample taken from axillary skin had three merits as follow:one,the results was more ideal.Incidence of vacuole of axillary sweat gland was higher than that of plantar sweat gland.Two,the results accorded with clinical practice.Axillary skin chould be taken in 30 minutes,it accorded with the characteristic of the Chinese herbs dispelling biao-zheng,and the diaphoretic function of MHT was rapid,but plantar skin chould be taken after two hours.Three,the slice could be made easily, and the quality of the slices was better.(2)MHT three dose (2g/kg, 4g/kg, 9g/kg)groups (counted by single Mahuang)exhibited their diaphoretic function in mice.The sweating volume was 0.240±0.079g,0.357±0.077g,0.556±0.082g,and the duct diameter of axillary sweatgland was 0.783±0.155um,2.235±0.273um,3.298±0.318um,respectively.They all presented dose-effect relationship.The correlation analysis showed that there was a significant positive correlation between the duct diameter(x) and the sweating volume(y)(r=0.927).R2=0.859 was elicited after the regression analysis,the regression equation was tested,F=231.428,p<0.001,it indicated that there was a statistical significance.The regression equation was y=-0.135+5.878x.(3)MHT three dose(4g/k^ 8g/kg^ 16g/kg) groups exhibited their diaphoretic function on rats,the duct diameter of axillary sweat gland was 1.22±0.09um,2.77±0.09um, 5.31±0.13um, respectively, and it presented a dose-effect relationship.2.Incidence of vacuole (% ) of MHT group,Mahuang+Guizhi+Xingren group, Mahuang +Guizhi+Gancao group,Mahuang+Xingren+Gancao group,Mahuang +Guizhi group,Mahuang+ Xingren group,Mahuang+Gancao group,Mahuang group, Guizhi group,Xingren group,Gancao group,Guizhi+Xingren+Gancao group,Xingren + Gancao group,and control group was 20.17±1.13,22.49±1.12,20.07±0.84, 1.92±0.70,22.79±1.09,8.80±0.58,1.94±0.56,9.47±1.15J.30±1.18,0.34±0.73,0.33±0.7 0,1.63±0.50,0.35±0.74,0.35±0.73,respectively.Incidence of vacuole of Mahuang group was highest in four single Chinese medice groups ,and Mahuang adding to Guizhi compatible group was the highest among fourteen groups. The diaphoretic function of compatible group including Mahuang was higher than that of compatible group no including Mahuang,respectively.The diaphoretic function became higher after adding to Guizhi,that had no change significantly after adding to Xingren, and that became weaker after adding to Gancao.3.(l)The suitable dosage observing diaphoresis in mice was MHT 9g/kg , at 30 minute after administration.(2)The sweating volume of MHT group,Mahuang+ Guizhi+Xingren group,Mahuang +Guizhi+Gancao group,Mahuang+Xingren+Gancao group,Mahuang+Guizhi group,Mahuang+ Xingren group,Mahuang+Gancao group, Mahuang group,Guizhi group,Xingren group,Gancao group,Guizhi+Xingren+ gahcao group , Xingren+Gancao group,and control group was 0.544±0.070g, 0.629±0.091g,0.496±0.084g,0.276±0.080g,0.685±0.074g,0.397±0.075g,0.318±0.048 g,0.440±0.090g,0.373±0.062g,0.124±0.044g,0.106±0.059g,0.248±0.053g,0.127±0.0 47g,0.099±0.059g,respectively.And their corresponding duct diameter of axillary sweat gland was 3.298±0.122um,3.535±0.142um,3.175±0.181um,0.653±0.057um, 3.778±0.183^m,3.053±0.127nm,0.685±0.106p.m,2.965±0.153nm50.763±0.104nm,0.5 15±0.104um,0.523±0.098um,0.635±0.060um,0.505±0.103^m,0.508±0.111 um,respec tively.The sweating volume of Mahuang group was strongest in four single Chinese medice groups ,and Mahuang adding to Guizhi compatible group was thestrongest among fourteen groups. The diaphoretic function of compatible group including Mahuang was stronger than that of compatible group no including Mahuang,respectively.The diaphoretic function became stronger after adding to Guizhi,that had no change significantly after adding to Xingren, and that became weaker after adding to Gancao.The correlation analysis showed that there was a significant positive correlation between the duct diameter(x) and the sweating volume(y). Pearson correlation coefficient was 0.87, p<0.001 (2-tailed),Spearman correlation coefficient was 0.96, p<0.001(2-tailed). R2=0.757 was elicited after the regression analysis,the regression equation was tested,F=429.819 ,p<0.001,it indicated that there was a statistical significance.The regression equation was y=0.117+0.127x.4.The anal temperature between MHT 9g/kg dosage group and control group hadn't difference significantly at 5 minutes,20 minutes and 25 minutes.But the anal temperature of MHT group was higher than that of control group at 10 minutes and 15minutes,lower at 30 minutes.Inside the MHT group,there were difference significantly between at 10 minutes and at 15 minutes(p<0.05), between at 15 minutes and at 10 minutes(p<0.001),between at 20 minutes and at 15 minutes (p<0.001). There weren't difference significantly between at 25 minutes and at 20 minutes(p>0.05), between at 30 minutes and at 25 minutes(p>0.05).In other words,the anal temperature of MHT group had an ascending trend at 5 minutes,began to rise at 10 minutes,arrived in highest point at 15 minutes,which had significant difference with other five time points(p<0.001),began to descend at 20 minutes,and at 30 minutes was lower than that of control group.5.Contrasted the left with the right,the diaphoretic function on left thenar sweat gland was disappeared,but the right was normal.6.MHT could expand auriclar artery and vein vessel, quicken the blood flow speed,and increase skin blood capacity. Mahuang could shrink auriclar artery and vein vessel ,Guizhi could expand auriclar artery and vein vessel, their effect on blood vessel were contrary.but both quickened the blood flow speed.The auricle microcirculation of different compatibility had no change significantly after adding to Xingren.Different compatibile groups had no change significantly in arterial and veinal blood vessel diameter after adding to Gancao, but they could quicken the blood flow speed.7. MHT manifested the diaphoretic function. NA at 0.39mg/kg dosage could restrain the diaphoretic function of MHT,andNA at 1.56mg/kg dosage could block the diaphoretic function .AD at O.lmg/kg dosage could restrain the diaphoreticfunction of MHT,and AD at 0.4mg/kg dosage could block the diaphoretic function .Although the duct of axillary sweat gland was expanded distinctly,the secretive phenomenon couldn't be seen.Phentolamine could speed the diaphoresis of MHT,and with the augment of the dosage of phentolamine,the secretion became more obvious.But propranolol restrained the diaphoretic function of MHT,and with the increasing of the propranolol dosage ,the diaphoretic function of MHT became weaker. Atenolol hadn't any impact on the diaphoretic function.8.MHT manifested the diaphoretic function.Atropine restrained the diaphoretic function of MHT,but couldn't block entirely. With the increasing of the atropine dosage ,the diaphoretic function of MHT became weaker.The diaphoretic function of MHT+atropine group was stronger than that of atropine+MHT group.Conclusions: l.In terms of observing the diaphoretic function of MHT on sweat gland,axilla is priority to plantar.Duct diameter of axillary sweat gland was regarded as a quantitative index to estimate sweating function.The results were credible, simple,intuitionistic,explict, and hardly influenced by environmental factors.This experiment could provide scientific, exact and available experimental method to traditional Chinese medicine about sweating method.2.Mahuang,Guizhi,Xingren,and Gancao played a role of principle drug .assistant drug,adjuvant drug and dispather drug in the docoction separately.lt embodied the compatible theories of the prescription thoroughly.3. There are a temperature ascend period at 5 minutes to 15 minutes in the process of the effects of MHT on anal temperature in normal mice.It showed the heat store in the body was increased,then the anal temperature descended as following diaphoresis.The effects of MHT on the set point in normal mice needed to be researched thoroughly.4.The diaphoretic function of MHT was.effective by nerve in rats.If sciatic nerve was severed,the diaphoretic function would disappear.5.The physiological basis about MHT's diaphoresis function lies in increasing skin blood capacity possibly.In the decoction,Guizhi assists Mahuang in diaphoresis. On the one hand ,Guizhi expands auriclar artery and vein vessel to decrease the shrinking effect of Mahuang, on the other hand, both quicken the blood flow speed, increase skin blood capacity to accommodate enough liquids that diaphoresis needs.6.The diaphoretic function of MHT was relative to adrenergic receptor.Activating a receptor, sympathetic nerve action enhanced, small artery in skin shrinked,the skin blood capacity reduced,and it could restrain the diaphoretic function of MHT,and blocking a receptor,the function was reverse.lt was testifiedthat (3 receptor was being in sweat gland.Activating P receptor ,small artery in skin expanded.But the duct of axillary sweat gland was expand activating a and P receptor simultaneously,the secretive phenomenon couldn't be seen.lt was obvious that in terms of diaphoresis,the restrained function of a receptor was more important than the expanded function of p receptor,even it was deciding.Blocking p receptor, small artery in skin shrinked.lt could restrain the diaphoretic function of MHT.The diaphoretic function might be effective mainly by activating other P receptor subtypes,not by activating Pi receptor.7.Atropine restrained the diaphoretic function of MHT,but couldn't block entirely.lt testified that the diaphoretic function of MHT was relative to activating M receptor,but wasn't an only approach.lt ascribed that adrenergic receptor and M receptor both belonged to G Protein-Coupled Rcecptor,the structures of these receptor were very resemble,they both were seven a -helices transmembrane segment receptor,their binding position lied in binding cranny formed by striding- membrance a-helices.Because of the heterogenous modulation,adrenalin not only could activate adrenergic receptor,but also could activate M receptor.Atropine was a M receptor competitive antagonist,even though at 0.4mg/kg dosage in mice,it couldn't block the diaphoretic function of MHT.Because sweat gland might be controlled by cholinergic and adrenergic nerve fibre simultaneously.When M receptor was blocked, MHT could exert the diaphoretic function by adrenergic nerve fibre.It was this reason that the difference order of administration of MHT and atropine resulted in the difference in diaphoresis. |
