Mechanism Of Kidney Deficiency And Liver Invasion And Pharmacological Study Of Drugs For Nourishing Kidney And Softening Liver—Study On The Pharmacodynamics Mechanism Of Zhenshao Chungan In The Treatment Of Menopausal Syndrome In Rats With Kidney Deficien

1.Theory ResearchDuring period of menopause,menstrual disorders occurred in women,accompanied by symptoms associated with menopause such as hot flush,sweating,and irritability.The syndrome was called "premenopausal and postmenopausal syndrome".In modern medicine,it was called "menopausal syndrome(MPS)".Kidney deficiency and liver invasion type is one of the common syndromes of MPS.In TCM,during the period of menopause,the essence of the kidneys is gradually weakening,the Tian gui will be absolutely ruined,the Chong and Ren channels will be empty,the essence and blood will be insufficient,the essence does not nourish liver,yin would not restrict yang,and yang is hyperactivity.Liver-qi disorders induces the liver-qi transforming into fire,which in turn consumes qi and yin,aggravating yin-blood deficiency.Therefore,Kidney deficiency is the root of the disease,and liver invasion is an important factor in the pathogenesis.As suggested in modern medicine,the pathogenesis of this disease is complex,which is guided by HPGA dysfunction,and also closely related to neurotransmitters,vasomotor factors,and free radical scavenging.2.Experimental ResearchObjectives: Rats with castration(ovarian ablation)and hot Chinese medicine(aconite and dry ginger)were used as research objects.Based on the behavioral characterization,the model was used to simulate the MPS kidney deficiency and liver invasion type and the therapeutic effect of nourishing kidney and softening liver method.By detecting endocrine levels and uterine morphology,the pathogenesis of MPS kidney deficiency and liver invasion type and the pharmacodynamic mechanism of nourishing kidney and softening liver were initially explored.Modern medicine was applied to reveal the scientific connotation of TCM treatment,promote the TCM treatment,and provide experimental basis for the development of new drugs.Methods: 1.3-month-old female SD rats were randomly divided into Control,OVX and OVX+FG groups.The skin temperature and water intake were monitored,EPM and resident intruder experiment were processed.The degree of agreement between the model and the MPS kidney deficiency and liver invasion type was evaluated by behavioral characterization.2.Zhenshao Chungan was given for the model rats.Based on the given dose,the model rats were divided into high,middle and low groups.Rats treated by Progynova and Kunbao Wan were used as positive controls.An equal amount of solvent(0.5% CMC-Na)was given to rats in the Model and Sham groups.After treatment for 28 d,body weight,skin temperature and core temperature were monitored.EPM and resident intruder experiment were processed.ELISA assay was used to detect E2,P,NO,ET-1,?-EP and ALLO levels in serum,hypothalamus Gn RH,pituitary FSH and LH levels.The pathological changes of uterus were observed by HE staining technique.Immunofluorescence and real-time PCR were used to detect the expression of estrogen receptor protein and related genes in uterus.Results: 1.Compared with the Control group,the skin temperature of OVX and OVX+FG group increased significantly(P<0.05).The OE% decreased significantly(P<0.05),and the mixed aggression behavior score increased significantly(P<0.05).Compared with the OVX group,the skin temperature of OVX+FG group raised even more(P<0.01),and the amount of water intake increased(P<0.01),the OE% and OT% were further reduced(P<0.01).2.Behavioral characterization: Compared with the Sham group,the weight of the Model group was significantly increased(P<0.01),OE% was significantly decreased(P<0.05),while OT% was decreased with no statistical difference.Compared with the Model group,the weight gain rate of the Progynova and High ZS group was significantly slower(P<0.05),the OE% and OT% of the High ZS group were significantly increased(P<0.05).There were no statistical differences in skin temperature,core temperature,and mixed aggressive behavior scores.3.Endocrine indexes: Compared with the Sham group,the serum levels of E2,ALLO,?-EP and NO in the Model group were significantly lower(P<0.05).Compared with the Model group,Progynova,Middle ZS and Low ZS group could significantly increase the E2 content(P<0.05).In the Kunbao Wan group,the ALLO content was significantly increased(P<0.01).High ZS and Middle ZS group could significantly increase the ?-EP content(P<0.05).NO content was significantly increased in the Kunbao Wan,High ZS and Low ZS group(P<0.05).There was no significant difference in serum P and ET-1 levels among groups.Compared with the Sham group,the hypothalamus Gn RH,pituitary FSH and LH contents in the Model group were significantly increased(P<0.05).Compared with the Model group,the content of hypothalamus Gn RH in the Kunbao Wan,Progynova,High ZS and Low ZS group was significantly decreased(P<0.01).The content of FSH in pituitary was decreased in the Progynova group(P<0.05).The LH content in pituitary was significantly decreased in all drug-administered groups(P<0.05).4.Uterine histological examination: Compared with the Sham group,the uterus of the Model group was significantly atrophied with decreased uterus index and thinner intima(P<0.01).Compared with the Model group,uterine index significantly increased in the Progynova and High ZS group(P<0.05),while intimal thickness was not changed.5.Uterine estrogen receptor protein and gene expression: Immunofluorescence results showed that compared with the Sham group,the levels of ER? and ER? in the Model group were significantly decreased(P<0.01).Compared with the Model group,Progynova,High ZS and Middle ZS group could significantly increase the ER? content(P<0.01),while the ER? content was significantly increased in all ZS groups(P<0.05).The real-time PCR results show that compared with the Sham group,the expression of ER?m RNA in the Model group was significantly decreased(P<0.01),but there was no significant difference in ER?m RNA.Compared with the Model group,the expression of ER?m RNA in the Progynova and High ZS group increased significantly(P<0.05).ER?m RNA increased in the High ZS and Middle ZS group,but with no statistical difference.Conclusions: 1.Rats with castration(ovarian ablation)and hot Chinese medicine(aconite and dry ginger)is a reliable model for studying the pathogenesis of this disease and the pharmacology of nourishing kidney and softening liver,which can simulate the symptoms of MPS such as hot flush and irritability.2.The disease may be caused by fluctuations in estrogen levels,HPGA dysfunction,visceral fat accumulation,changes in neurotransmitters and vasomotor factors level during menopause.3.After treatment with Zhenshao Chungan,the weight gain rate of the model rats slowed down,the uterus index increased,the OE% of OT% was elevated,and the anxiety improved,which proved that the treatment of nourishing kidney and softening liver was effective.4.After treatment with Zhenshao Chungan,the levels of serum E2,?-EP and NO was increased,and the contents of central Gn RH,LH and FSH was decreased.Based on above results,it was speculated that nourishing kidney and softening liver method might regulate the disordered HPGA,and balance the ?-EP and NO levels to improve the symptoms of MPS.5.After treatment with Zhenshao Chungan,both E2 content in the serum and the ERs expression in the uterus increased.Thereby,the nourishing kidney and softening liver method may play a therapeutic role by simultaneously increasing the ligand and receptor content,delaying the regression of the uterus.In addition,the treatment did not cause endometrial thickening,evading the side effects of hormone therapy.