| Objective: 1.In order to find a simple and ideal Uterine Leiomyoma(UL)animal model for research,according to the traditional Chinese and western medicine pathogenesis of UL,the UL models with different methods were prepared and compared with various aspects,such as organ coefficient,uterine size,uterine pathology,estrogen,progesterone and receptor concentration in the UL rats.2.The method of exerting estrogen and progesterone combined with multi-factor stimulation was used to establish the rat modelof qi stagnation and blood stasis disease combined with UL,which was comprehensively evaluated from the "cause","phenomenon","syndrome" and "treatment".Study on the pharmacodynamic effect of Sanleng-Ezhu Component Compatibility was observed through end-points which include organ coefficient of the thymus,spleen and uterus,uterine pathology and morphology,blood rheology,IL-1β,TNF-α,GNRH,FSH,LH,E2,P,ER,PR and so on.It was to explore the intervention mechanism of Sanleng-Ezhu component compatibility,and to provide the basis for clinical treatment.Methods: 1.The rats were randomly divided into control group,short-time model group,combining disease with syndrome group,long-time model group,qi stagnation and blood stasis group.Every Monday,Wednesday,Friday,the long-time model group was gavaged with diethylstilbestrol 0.167mg/kg,a week for 3 times,and each rat was injected intramuscularly with Progesterone Injection 1.0mg every Sunday,for 20 weeks.Short-time model group and Combining disease with syndrome group were gauged with diethylstilbestrol 1.35mg/kg,and Every Monday,Wednesday,Friday,each rat was injected intramuscularly with Progesterone Injection 1.0mg,for 5 weeks.The qi stagnation and Combining disease with syndrome group were subcutaneously injected with Adrenaline Hydrochlaride Injection 0.9mg/kg·d-1,then the following chronic unpredictable stilmulation was carried out randomly.((1)60d B noise for 3h,(2)Day reversal,(3)Swimming in the 5~10℃ water for 4 minutes,(4)Hanging tail 10 min,(5)Bake for 10 minutes at 50℃.A daily stimulation was done,and each stimulus was not less than 2 times within 2 weeks.The rats were randomly divided into control group,model group,positive control group,S-E high dose group,S-E middle dose group and S-E low dose group.In addition to the control group,the other groups were prepared model according to the combining disease with syndrome group,then 66.7%,33.3%,16.7% S-E liquid and positive drug were gavaged for continuous 5 weeks,the control group and model group were gavaged distilled water.Results: 1.The phenomenon of rats’ bleak color,ruffled fur,epilation,lassitude was appeared after preparing model.At the end of the experiment,the uterine appearance of the obvious nodules,cysts and swelling was shown in the other group especially in the long-time model group except for qi stagnation and blood stasis group.Pathological changes of muscle cells and inflammatory cell infiltration were observed,and the uterine smooth muscle cells’ degeneration and proliferation were obviously shown in the combining disease with syndrome group and long-time model group.Comparison of estrogen and progesterone,the model’s P in the serum and E2,P,ER,PR in the uterus and ovary were significantly increased,especially in the combining disease with syndrome group(P < 0.01).From the comparison of modeling rate,the rate of short-time model group,combining disease with syndrome group and long-time model group was successively 42%,58% and 67%.According to the overall analysis of the modeling methods,modeling time,modeling rate and other data,the combining disease with syndrome group had a greater advantage than other groups,an alternative UL model tool had been provided.2、The qi stagnation and blood stasis of UL animal model was evaluated from four aspects.Firstly,etiological analysis of qi stagnation and blood stasis syndrome was considered,the rats’ state of "annoyance","anger" and "lush" was formed from emotions,external environment and endocrine.Secondly,the biological characters such as diet reduction,black-purple nail and tail,epilation,abscess of chest and flank,arched backs,curled up,reduced activity,irritability,were conformed to the symptoms of qi stagnation and blood stasis.Thirdly,the pathomechanism study showed the uterine anatomy of nodules,cysts and swelling,and the uterine pathology of the uterine mucosa epithelial cells vacuolar degeneration,smooth muscle cell degeneration,disorder and proliferation,the increased hemorheology parameters of model rats.Finally,the syndrome was proved by the formula.The uterine morphology and the hemorheology parameters of UL rats had been markedly improved by treating of San Leng-EZhu.The results showed that San Leng-EZhu could improve the status of qi stagnation and blood stasis.3、Compared with the model group,the FSH and LH concentration in the pituitary gland of S-E high group and positive drug group were significantly decreased.The E2,P,ER,PR concentration in uterus and ovary of S-E high,middle,low group and positive drug group were significantly decreased,the difference was statistically significant(P < 0.05).The results showed that S-E could decrease the concentration of FSH and LH in the pituitary and adjust the concentration of E2 and P to decrease the stimulation of estrogen and progesterone on endometrium.The results showed that S-E could adjust the HPG by regulating the content of estrogen and progesterone and the expression of the receptor,and affecting the secretion of FSH and LH in the pituitary gland,and then inhibiting secretion of estrogen and progesterone.S-E could decrease the level of TNF-α in serum and IL-1β,TNF-α in uterus,improve the inflammatory reaction and self-immunocompetence in the UL rats,and promote recovery of ovarian and uterine function.So S-E could promote the negative feedback effect of estrogen and progesterone on hypothalamic-pituitary through the influence of neuroendocrine and immune system.Conclusion: 1.The animal model of uterine leiomyoma was successfully prepared by exerting estrogen and progesterone.2.The UL rat model combined with qi stagnation and blood stasis was successfully established by exerting estrogen,progesterone and multi-factor stimulation,which was evaluated from four aspects: "cause","phenomenon","syndrome" and "treatment".The results showed that the model was stable,the modeling rate was comparatively high,and the UL modeling time could be significantly shorten,which provided an alternative model tool for the study of uterine leiomyoma.3.The intervention mechanism of Sanleng-Ezhu component compatibility on the UL rats was demonstrated from the following aspects:firstly,Sanleng-Ezhu could improve the hemorheology indexes and relieve qi stagnation and blood stasis of model rats by activating blood circulation to dissipate blood stasis;Secondly,it could innhibit the occurrence and development of UL through the intervention of HPG axis and the estrogen and progesterone secretion.At last,Sanleng-Ezhu component compatibility could promote the function of the rats’ uterus and ovary by improving the UL rats’ immune ability. |
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