| A large number of studies have shown that the modern connotation of TCM syndrome involves the pathophysiological changes of multiple systems.liver depression syndrome involves number of systems abnormalities including nervous,endocrine,immune,digestive,blood system.With the background of liver in traditional Chinese medicine and moderrn reproductive system,and clinical liver depression syndrome often come with reproductive disease,it is suggested that liver depression syndrome may involve the abnormality of reproductive system.Because of the previous researchs of rat model with liver depression syndrome,In this time the changes of reproductive system were observed dynamically on this model.Based on the traditional Chinese medicin’s correlation of formula and syndrome principle,Compared with the effects of Chinese medicine Shugan formula(Chaihu-Shugan Powder),Bushen Fang(Wuzi-Yanzong Pills)and Fluoxetine hydrochloride on the model of liver depression syndrome and the prevention effects on gonadal function of that model;on this basis,from the molecular level to further explore the drugs protection mechanism on rat testicular injury in liver depression syndrome.Researceh 1 Gonadal functions and changes in rats with liver depression syndromeMethods:Sixty healthy male Wistar rats weighing 200 ± 20g were randomly divided into normal control group(n= 30)and model group(n=30).The model group was restructured liver depression syndrome according to the method of shaking for 3 hours and 4 weeks.The normal control group was not treated.Ten rats were randomly selected from the normal group and the model group at the end of the second week,the third week and the fourth week of the experiment.The rats were sacrificed by intraperitoneal anesthesia with 10%chloral hydrate(0.30ml/100g)And take the testis,hypothalamus and other organizations.(1)Indicator of syndrome:5-HT of hypothalamus,serum Norepinephrine(NE),serum cortisol(CORT);(2)gonadal function indicators:1)serum hormone:gonadotropin releasing hormone(GnRH),luteinizing hormone(FSH),follicle stimulating hormone(LH),prolactin(PRL),testosterone(T),estradiol(E2);2)gonadal organ:testicular wet weight and organ index,testicular HE observation.The general appearance of the rats was observed and recorded every week include behavior status,activity level,emotional response,diet status,and body weight.Results:(1)Changes of TCM syndromes in rats with liver depression syndrome:Compared with the normal group,the behavioral status,the degree of activity and the emotional response of the rats in the model group increased in2nd-4th week(P<0.0l).The levels of 5-HT in the hypothalamus were significantly decreased at the end of2nd week and significantly increased at 3rd and 4th weeks(p<0.01).The levels of 5-HT were significantly lower than those in the control group(P<0.01),NE and Cort were significantly increased in2nd-4th week(p<0.01 or p<0.05).(2)Changes of gonadal function in rats with liver depression:Compared with the normal group,the model group’s GnRH was increased inthe 2nd and 3rd week(P<0.01),decreased at 4th week.The levels of FSH and LH were significantly decreased at 2nd-4th week(p<0.01).The levels of PRL and E2 were significantly increased at 2nd-4th week(P<0.01).The testicular weight and pathological damage score were significantly decreased at 2nd weeks and 4th weeks(p<0.01 or p<0.05).Conclusion:liver depression model rats which useing the method of bound stress 4 weeks restructured has showed similar to the appearance of TCM liver depression and similar changes in laboratory indicators,accompanied gonadal axis function and testicular injury.With the extension of model time,the model rats showed that GnRH increased at first and then decreased,andT,LH and FSH/PRL and E2 decreased or increased apartly,and the pathological damage of testis gradually increased.Suggesting that the hypothalamus may be the main parts of the adjustment of the gonadal axis in the development of liver depression syndrome.Reserash 2 The comparison of Shugan formula and Bushen formula in the mechanisms of HPG axis of liver-depressed male ratsMethods:69 healthy male Wistar rats weighing 200 ± 20g were randomly divided into normal control group(n=12)and model group(n=57).Normal control group was not treated,the model group rats accordance with the above methods to copy liver depression model.The rats in the model group were randomly divided into treated groups,Chaihu Shugan High dose group(n=12)and Chaihu Shugan low dose group(n=12).Western medicine group(n=9),Bushen group(n=12).Among them,Chaihu Shugan Powder was divided into single dose of 1.24g/kg and 0.31g/kg(equivalent to 2 times and 0.5 times of adult dosage of 70kg human body weight).The western medicine group was treated with 1.8mg/kg(equivalent to 70kg ofhuman body weight)fluoxetine hydrochloride,Bushen group treated with 1.08g/kg(equivalent to 70kg human body weight)Wuzi-Yanzong Pills.One time per day,continuous gavage for two weeks.laboratory Indicators:(1)appearance behavior indicators and open field test;syndrome-related laboratory indicators:hypothalamus 5-HT,serum NE,E,Ach,CORT.(2)HPG axis related indicators:serum GnRH,FSH,LH,T,E2,PRL;testicular wet weight and organ index,testicular HE pathological observation.(3)SOD,MDA,CytC and 8-OHdG of testicular mitochondrial;2)testicular mitochondrial PKA,ERK,STAR and Cypllal.Results:(1)Changes in syndromes:1)Behavioral changes:Compared with the normal group,the scores of the rats in the model group were significantly increased in2nd-4th weeks,and the body weight was significantly decreased(p<0.01),at the 2nd week The number of crawl and crawling mileage and the residence time of the central grid in the open field test increased significantly(p<0.01).and the 4th week The number of crawl and crawling mileage decreased and the residence time of the central grid in the open field test increased significantly(p<0.01).Compared with the model group,the scores of the high dose group and western medicine group were significantly decreased(p<0.01),the number of crawl and crawling mileage increased significantly,and the residence time in the central grid was significantly decreased(p<0.01),And the weight of High dose group was significantly increased,There was no significant difference between the western medicine group and the Bushen group.2)Syndrome-related laboratory indicators:Compared with the normal group,the levels of 5-HT,serum NE and CORT of the model group were significantly increased(p<0.01),and Ach was significantly decreased(p<0.01).Compared with the model group,the levels of 5-HT,serum NE and CORT of the High dose group were significantly decreased(p<0.01),The low dose group Ach was significantly increased(p<0.01),NE was significantly decreased(P<0.01).The levels of NE,E and CORT were significantly decreased in Bushen group and western medicine group.(2)HPG axis correlation indicators:Compared with the normal group,the levels of GnRH,LH and T in the blood of model group were significantly decreased(p<0.01),PRL and E2nd were significantly increased,the weight of testis was significantly decreased,pathological damage score were significantly increased(p<0.01).Compared with the model group,the levels of GnRH,LH and T in the High dose group were significantly increased(p<0.01 or p<0.05),PRL and E2were significantly decreased,the weight of testis was significantly increased,and the score of pathological damage was significantly decreased(P<0.0l or p<0.05).In low dose group the levels of PRL and E2were significantly lower than those in the control group(P<0.01).In Bushen group GnRH Significantly increased,E2 significantly decreased(p<0.01 or p<0.05).(3)Compared with the normal group,the mitochondrial MDA,CytC and 8-OHdG were significantly higher than those of the normal group(p<0.01 or p<0.05).Compared with the model group,the high dose group mitochondrial MDA,CytC and 8-OHdG in the testis were significantly decreased and the SOD level was significantly increased(p<0.01 or p<0.05).the low dose group mitochondrial MDA of the testis decreased significantly(P<0.01 or p<0.05).The mitochondrial SOD of testis was significantly increased in western medicine group(p<0.05).2)Compared with the normal group,PKA,ERK,STAR and Cypllal were significantly decreased in the model group(p<0.01 or p<0.05).Compared with the model group,PKA,ERK,STAR and Cypllal were significantly increased in the high dose group(p<0.01 or p<0.05).The low dose group PKA and STAR increased significantly(p<0.01 or P<0.05).The western medicine group PKA of the testis was significantly increased(P<0.01).Conclusion:the high or low dose Chaihu Shugan Powder and western medicine(fluoxetine)have different effects on the behavior of rats with liver depression syndrome,and the effect of high dose Chaihu Shugan Powder is most obvious,Bushen formula(Wuzi Yanzong pill)can not achieve this effect.Chaihu Shugan Powder has a good effect on the HPG axis of the model rats and relieves the pathological damage of the testes in the model rats.The testicular protective effect may because of it can reduce mitochondrial oxidative stress injury and protect of testosterone production signal transduction pathway.Bushen Formula and western medicine group only have a certain improvementon part of the indicators.The results of this study provide a certain biological evidence of the correlation of Chaihu-Shugan Powderto liver depression syndrome,and give a help to understand the pathophysiological mechanism of the HPG axis system changes in liver depression syndrome,Proved that it is reasonable for choose different prescriptions for different Syndromes as all have known in Traditional Chinese medicine. |
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