| Objective: SD female rats are used as research objects to establish an animal model of menopausal syndrome with kidney-yin deficiency syndrome,and to obtain the general behavior,tissue morphology,sex hormones and central neurotransmitters of experimental animals related to changes in hypothalamus-pituitary-gonadal axis function.The experimental data are compared with those of non-menopausal normal young adults to explore the biological mechanism of "hot flashes" in menopausal kidney-yin deficiency syndrome.In addition,the changes in the external environment temperature are used as the triggering factors of "hot flashes",analyzes the differences in the performance of "hot flashes" in different seasons,and explores the biological mechanism of changes in the external temperature and environment to promote "hot flashes",with a view to exploring the mechanism of menopausal "hot flashes" from both internal and external levels.Methods:100 sexually mature female Spreque-Dawley(SD)rats at 12 weeks of age,weighing(220 ± 20)g,SPF grade.100 12-week-old sexually mature female SD rats were randomly divided into 5 groups: menopausal syndrome disease model group,disease treatment group,menopausal syndrome kidney yin deficiency syndrome disease model group,disease syndrome treatment group and blank group.Each group has 20 animals,the same experiment was performed in the summer and winter seasons.The summer and winter experiment time was around in the 4 weeks before and after the summer solstice and winter solstice,the summer experiment is from May 24 to July 19,2018,the specific time of the winter experiment is November 24,2018 to January 19,2019.Observation indicators: 1.General conditions: rat weight,hair,diet,water intake,stool,activity,sleep,emotion,etc.;2.Tail temperature of rats;3.Serum sex hormone levels(E2,FSH,LH,GnRH);4.Neurotransmitter concentration: neurotransmitter level(5-HT)in the hypothalamus,norepinephrine(NE)in serum;5.Uterine pathological section,uterine coefficient(uterine coefficient = uterine wet mass / body weight).Results: 1.General situationExcept that the hair color and hair quality of the rats in the blank group did not change too much during the whole course of the experiment,the hair color of the other groups of rats showed yellowishness,poor gloss,and softness and lack of touch;Compared with the rats in the blank group,the other rats gained weight,showed slightly agitated behavior.The rats in the disease syndrome model group also showed red tongue tips,increased drinking water,and dry stool.The rats in the disease treatment group and the disease syndrome treatment group were still in good spirits and their behavioral agitation was improved.Death of rats: 3 rats died due to improper gastric administration,including 2 rats of disease syndrome treatment group in summer and 1 rat of disease syndrome treatment group in winter.2.Rat tail temperatureDuring the whole experiment,the tail temperature of rats showed a certain fluctuation trend with the change of outside air temperature: the outside temperature in summer(24.47 ± 1.19)℃,the tail temperature of rats in the blank group(29.17 ± 1.64)℃,and the outside temperature in winter(9.85 ± 3.60)℃,and the tail temperature of the rats in the blank group was(18.26 ± 1.19)℃.Ovariectomized rats had a higher tail temperature since the ovariectomy than rats in the blank group(average temperature of rat tail temperature in the first week after ovariectomy in summer: blank group(29.49 ± 0.49)℃,disease model group(31.27 ± 0.32)℃;mean temperature of rats tail temperature at 1 week after ovariectomy in winter: blank group(19.92 ± 0.24)℃,disease model group(21.66 ± 0.25)℃).The tail temperature of disease syndrome model rats was higher than the disease model group(in the third week after ovariectomy in summer: disease syndrome model(30.73±0.38)℃,disease model group(29.74±0.45)℃).Moreover,the trend of tail temperature rise in each group of ovariectomized rats would be close to the tail temperature fluctuation level of the blank group at 5-6 weeks after ovariectomy.After the treatment of estradiol valerate and zuogui pill,the tail temperature of rats in the disease treatment group and the disease syndrome treatment group showed a certain trend of decline,and gradually recovered to the tail temperature of rats in the blank group over time.In the winter experiment,the tail temperature of each group of ovariectomized rats,compared with the tail temperature of the blank group,its fluctuation tendency and maintenance time(except the duration of the disease model group)were smaller than the summer group experiment.From summer to winter,with the decrease of the external temperature,the overall tail temperature of the rats also decreased,and the fluctuation of tail temperature of the rats in each group of ovariectomy also decreased.Throughout the experiment,the high tail temperature of the rats in each group of ovariectomy would eventually return to the tail temperature of the normal blank group over time.3.Changes in sex hormone levelsIn the whole experiments,when the levels of Estradiol in serum of ovariectomized rats compared with the blank group(in summer the blank group 380.01±22.99 pg/ml;disease model group 42.79±8.40 pg/ml),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(42.79±8.40 pg/ml)and disease treatment group(94.20±9.74 pg/ml);there was a statistical significance in difference between the disease syndrome model group(42.76±8.30 pg/ml)and disease syndrome treatment group(100.15±12.39 pg/ml).There was no statistically significant difference between the summer and winter in the each group of Estradiol in serum.In the whole experiments,when the levels of Follicle stimulating hormone in serum of ovariectomized rats compared with the blank group(in summer the blank group 10.66±1.22 mIU/mL;disease model group 19.14±1.47 mIU/mL),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(19.14±1.47 mIU/mL)and disease treatment group(15.23±1.40 mIU/mL);there was a statistical significance in difference between the disease syndrome model group(21.40±1.87 mIU/mL)and disease syndrome treatment group(14.84±1.50 mIU/mL).There was no statistically significant difference between the summer and winter in the each group of Follicle stimulating hormone in serum.In the whole experiments,when the levels of Luteinizing hormone in serum of ovariectomized rats compared with the blank group(in summer the blank group 11.02±1.24 IU/L;disease model group 18.26±1.16 IU/L),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(18.26±1.16 IU/L)and disease treatment group(14.35±1.08 IU/L);there was a statistical significance in difference between the disease syndrome model group(19.82±1.12 IU/L)and disease syndrome treatment group(14.74±1.03 IU/L).There was no statistically significant difference between the summer and winter in the each group of Luteinizing hormone in serum.In the whole experiments,when the levels of Gonadotropin-releasing hormone in serum of ovariectomized rats compared with the blank group(in summer the blank group 219.34±15.98 pg/ml;disease model group 341.48±22.41 pg/ml),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(341.48±22.41 pg/ml)and disease treatment group(302.26±22.41 pg/ml);there was a statistical significance in difference between the disease syndrome model group(352.25±19.88 pg/ml)and disease syndrome treatment group(286.61±17.03 pg/ml).There was no statistically significant difference between the summer and winter in the each group of Gonadotropin-releasing hormone in serum.4.Changes in neurotransmitter concentrationIn the whole experiments,when the levels of 5-hydroxytryptamine in Hypothalamic homogenate of ovariectomized rats compared with the blank group(in summer the blank group 437.25±28.07 ng/ml;disease model group 295.50±25.71 ng/ml),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(295.50±25.71 ng/ml)and disease treatment group(376.38±28.52 ng/ml);there was a statistical significance in difference between the disease syndrome model group(312.13±24.95 ng/ml)and disease syndrome treatment group(377.52±25.07 ng/ml).There was no statistically significant difference between the summer and winter in the each group of 5-hydroxytryptamine in Hypothalamic homogenate.In the whole experiments,when the levels of norepinephrine in serum of ovariectomized rats compared with the blank group(in summer the blank group 230.11±24.24 pg/ml;disease model group 300.35 ± 29.46 pg/ml),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(300.35±29.46 pg/ml)and disease treatment group(260.38±21.26 pg/ml);there was a statistical significance in difference between the disease syndrome model group(289.57±27.70 pg/ml)and disease syndrome treatment group(256.14±23.17 pg/ml).There was no statistically significant difference between the summer and winter in the each group of norepinephrine in serum.5.Difference of uterine coefficient and morphology of uterusIn the whole experiments,when the uterine coefficient of ovariectomized rats compared with the blank group(in summer the blank group 1.71±0.12 mg/g;disease model group 0.31±0.05 mg/g),there was a statistical significance in difference;there was a statistical significance in difference between the menopausal syndrome disease model group(0.31±0.05 mg/g)and disease treatment group(0.39±0.07 mg/g);there was a statistical significance in difference between the disease syndrome model group(0.35±0.04 mg/g)and disease syndrome treatment group(0.42±0.07 mg/g).There was no statistically significant difference between the summer and winter in the each group of norepinephrine in serum.In the experiment,estrogen and progesterone levels in the body of rats decreased after ovarian excision,resulting in smaller uterine atrophy and thicker endometrium.Partial tissue edema or hyperemia;There were more glands and capillaries in utero in the disease treatment group and disease syndrome treatment group than in the disease model group and disease syndrome model group.Conclusion:1.Evaluation of animal model of menopausal syndrome with kidney-yin deficiency syndromeFemale mature SD rats were treated with ovariectomy.Vaginal smear was performed for 5 consecutive days from 5 days after surgery,no emotional response,successful building the animal model of menopause syndrome.Then gave the rats lavage treatment of 2 weeks Fuzi-dry ginger liquid which was used to administer the Yin-deficiency medicine by heat Chinese medicine.After 2 weeks,the levels of estradiol in the serum of rats reduced,and the levels of follicle stimulating hormone,luteinizing hormone and gonadotropin releasing hormone elevated;uterine volume atrophy,uterine pathological section pale red,local tissue edema,reduced capillaries;rat hair color yellowish,dark gloss,less compliant,red tongue tip color,increased drinking water,dry stool,increased anal temperature And restless.All of the above meet the requirements of the animal model of menopausal syndrome with kidney-yin deficiency syndrome,so female mature rats were given Fuzi-dry ginger liquid for 2 weeks after ovariectomy to prepare rats with menopausal syndrome with kidney-yin deficiency syndrome model method was effective.2.After successful modeling of the SD female rat model of menopausal syndrome with kidney-yin deficiency syndrome,the tail temperature rose and returned to the level consistent of the normal rat tail temperature after a period of time;speculates that the symptoms of hot flashes in menopausal syndrome may occur after ovariectomy,SD female rats have lower serum estradiol levels,which in turn affects the hypothalamic gonadotropin-releasing hormone neurons to release gonadotropin-releasing hormone,leading to the changes in the concentration of other neurotransmitters in the hypothalamus,narrowing of the thermoneutral zone,and changes in the temperature setting point.Therefore,the mechanism of hot flashes in menopausal syndrome may be related to the changes in the function of hypothalamus-pituitary-gonad axis and vasomotor dysfunction in the menopausal body.3.From summer to winter,the external ambient temperature decreases,and the trend and duration of the increase in tail temperature in the model group of menopausal syndrome with kidney-yin deficiency syndrome were reduced,however,no statistical significance was found in the difference between summer and winter in each group.It is speculated that the change in environmental temperature may be the menopausal syndrome with kidney-yin deficiency Prove the predisposing factors of the fluctuation of tail temperature in menopausal syndrome with kidney-yin deficiency syndrome model rats.4.The tail temperature of rats in the model group of menopausal syndrome with kidney-yin deficiency syndrome was higher than the simple disease model group.After the intervention of Zuogui pills,the fluctuation trend of tail temperature was relieved.In addition,the concentration of estradiol in the serum of disease syndrome treatment group rats increased,the level of the follicle stimulating hormone,luteinizing hormone,gonadotropin-releasing hormone had decreased.In this experiment,the use of Zuogui Pills to nourish the kidney and yin and fill the essence to intervene syndrome intervention can relieve the hot flashes by adjusting the changes of serum hormone levels and neurotransmitter concentrations in rats with menopausal syndrome and kidney yin deficiency syndrome,to some extent. |
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