| Objective:To research the three effective components in traditional Chinese medicine Panax pseudoginseng. To extract the effective parts (notoginsenoside, panax notoginseng flavonoids, panax notoginseng acid) and the mixture of three effective parts and to detect the PAI-1, TF, TFPI-2, TGF-β1changes in the underage rats'endometrium under the continuous external estrogen intervention in the full period. To explore the changes of the endometrial cells factors of underage rats under the effect of eogenous etrogen from the perspectives of fibrinolysis, coagulation factor and extracellular matrix, to analyze the relationship between the stability of endometrium and uerine bleeding repair under the influence of Panax pseudoginseng to eogenous estrogen as well as the relationship among the three kinds of effective parts and to investigate the scientific connotation of the "dissolving stasis and reducing blooding" of Panax pseudoginseng.Method:1. Divide120underage female rats into the bland group (40) and the model group (80) at random. Give normal saline irrigation to the blank group and exogenous estrogen (estradiol valerate) suspension irrigation to the model group for3weeks respectively. Observe the changes of endometrial pathological morphology and detect the changes of PAI-1, TF, TFPI-2TGF-β1in endometrium by using Immunohistochemistry and In Situ Hybridization.2.After a full period (3weeks) of continuous exogenous estrogen intervention, divide the rats without sexual cycle into6groups at random, namely the model group, the continuous estrogen group (hereinafter referred to as the estrogen group), the notoginsenoside group, the panax notoginseng flavonoids group, the panax notoginseng acid group, the panax notoginseng mixed group of effective parts (hereinafter referred to as the panax notoginseng mixed group). Each group were given corresponding normal saline, estrogen, notoginsenoside, panax notoginseng flavonoids, panax notoginseng acid and the mixture of three effective parts respectively for stomach irrigation for2weeks. Put8rats to death each weekend. Detect the changes of PAI-1, TF, TFPI-2, TGF-β1in endometrium by using Immunohistochemistry and In Situ Hybridization.Results:1. Compared with the blank group, the endometrial glands and stroma of the model group have hyperplasia in the first three weeks. In the end of the third week, all expression of rats'endometrial glands TF TFmRNA, PAI-1, PAI-1mRNA, TGF-β1of the model group rises (P<0.05, P<0.01) compared with that of the blank group, whereas its expression of TFPI-2, TFPI-2mRNA drops compared with the blank group.2. In the end of the fifth week, the PAI-1, PAI-1mRNA expression of the notoginsenoside group, the panax notoginseng acid group drops compared with the model group (P<0.01), the PAI-1, PAI-1mRNA expression of the panax notoginseng mixed group drops compared with the notoginsenoside group (P<0.05)3. In the end of the fourth week, the TF, TFmRNA expression of the panax notoginseng mixed group drops (P<0.05) compared with the model group. In the fifth week, the TF, TFmRNA expression of the notoginsenoside group, the panax notoginseng flavonoids group, the panax notoginseng acid group and the panax notoginseng mixed group drops (P<0.01, P<0.05)compared with the model group and the estrogen group, the TF, TFmRNA expression of notoginsenoside group and the panax notoginseng mixed group drops (P<0.01, P<0.05) compared with the panax notoginseng flavonoids group.4. In the end of the fifth weekend, the TFPI-2expression of the estrogen group drops (P<0.05), the TFPI-2mRNA expression of the notoginsenoside group, the panax notoginseng flavonoids group, the panax notoginseng acid group and the panax notoginseng mixed group rises (P<0.01, P<0.05) compared with the model group. In the end of the fourth and the fifth week, the TFPI-2, TFPI-2mRNA expression of the panax notoginseng acid group and the panax notoginseng mixed group rises (P<0.05, P<0.05) compared with the notoginsenoside group, the panax notoginseng flavonoids group.5. In the fourth week, the TGF-β1, TGF-β1mRNA expression of the notoginsenoside group, the panax notoginseng flavonoids group, the panax notoginseng acid group and the panax notoginseng mixed group drops compared with the model group (P<0.05, P<0.01), the TGF-β1mRNA expression of the panax notoginseng mixed group rises (P<0.05) compared with the the panax notoginseng flavonoids group. In the fifth week, the TGF-β1and/or TGF-β1mRNA expression of the notoginsenoside group, the panax notoginseng flavonoids group, the panax notoginseng acid group and the panax notoginseng mixed group drops (P<0.05, P<0.01) compared with the model group.Conclusion:1.After a full period of continuous exogenous estrogen intervention to the underage female rats, the majority of their endometrium have hyperplasia, the model of estrogen intervention to rats' hystera is successfully built. Based on the reasonable dosage of exogenous estrogen intervention, the pathological form of rats' endometrium changes, suggesting that it is similar to anovulatory dysfunctional uterine bleeding caused by a single estrogen.2.The research shows that all the three active parts of panax notoginseng can reduce the drops of rats'endometrial tissue PAI-1expression under a full period of continuous exogenous estrogen intervention. The effect of mixture of the three active parts of panax notoginseng is remarkable to invigorate the circulation of blood and stop bleeding by starting fibrinolytic system, this probably is one of the hemostatic mechanism of panax notoginseng's active parts curing anovulatory dysfunctional uterine bleeding.3.The expression of notoginsenoside and the mixture of effective parts of panax notoginseng to rats'endometrial tissue TF under a full period of continuous exogenous estrogen intervention, its possible mechanism is inhibition of tissue factor, blocking the activation of the coagulation system, reducing the inflammatory response, and achieving the purpose of invigorate the circulation of blood and stop bleeding.4.All the three active parts of panax notoginseng can increase rats' endometrial tissue TFPI-2expression under a full period of continuous exogenous estrogen intervention, while the panax notoginseng acid and the mixture of effective parts of panax notoginseng effect remarkably, which mechanism may strengthen the endometrial extracellular matrix to maintain the stability of the endometrium, thus reduce endometrial disintegration bleeding, and is good for endometrial repair and remodeling.5.All the three active parts of panax notoginseng can reduce rats' endometrial tissue TGF-β1expression under a full period of continuous exogenous estrogen intervention, while the panax notoginseng flavonoids and the mixture of effective parts of panax notoginseng effect remarkably, suggesting panax notoginseng flavonoids and the mixture of effective parts of panax notoginseng's treatment of uterine bleeding may be related to inhibition of TGF-β1over-expression, through anti-inflammatory, while improving microcirculation, promoting the degradation of extracellular matrix, acting the role of eliminating necrotic tissues and promoting granulation, which may be another mechanism of anti-inflammatory activity of the active parts of panax notoginseng.6.This research discovers that the mixture of3active parts of panax notoginseng perform the best in hem ostaticmechanism of rats' endometrium under a full period of continuous exogenous estrogen intervention, panax notoginseng, as the Hemostatic Rejuvenation, its3active parts are coordinated and indivisible. And this state is also consistent with hemostaticmechanism of dysfunctional uterine bleeding.
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