| 1.Experimental research:(1)Objective:By using the representative prescription of Warm Kidney and Remove Blood Stasis-HupoSan, to explore the molecular mechanism of the effect of Warming kidney and Removing Blood Stasis on EMs model rats, that is, the effect of MAPK signaling pathway and nuclear transcription factor kappa B on the endometrial tissue. To clarify the mechanism of the effect of the key steps in the treatment of endometrial adhesion, invasion and planting with the method of Warming Kidney and Removing Blood Stasis, and further to reveal the material basis and biological connotation of traditional Chinese medicine in the treatment of EMs.(2)Methods:The EMs rat model was constructed by transplanting the mature female SD rats, which were not mated. The rats were randomly divided into normal control group, model group,HuPoSan high, medium, low dose group, danazol group, and 15 in each group. After 1 weeks of modeling, the rats were given drugs for 3 weeks. Normal control group and endometriosis disease model group were given the same volume of normal saline. HuPoSan high, medium and low dose group:given the normal dosage of 20 times,10 times and 5 times respectively HuPoSan effective component water solution, Danazol group was given 10 times the normal amount of danazol effective component water solution. The growth of ectopic endometrium was observed at 4 weeks after modeling. The successful rats were respectively taken from the eutopic and the ectopic endometrial tissue, and normal tissue was taken from the normal group. By using the transmission electron microscope observated the ultrastructure of endometrial tissue, and the RT-PCR method was used to determine the expression of ERK, p38MAPK, NF-nuclear factor kappa mRNA B in endometrial tissues.(3)Results:①Transmission electron microscopy showed that the eutopic and ectopic endome-trial cell of endometriosis disease model group were active proliferation, such as mitochondria, rough endoplasmic reticulum, multi nuclear phenomena. The ectopic endometrial cells of Hu PoSan and the danazole group showed different degrees of cell injury and apoptosis, such as cell vacuolar degeneration, microvilli shortage and lack of nuclei deformity, core rim crest, etc..②The expression of ERK1/2, p38MAPK, NF-κB mRNA in disease model group was higher than that in normal control group, and the expression in ectopic endometrium was higher than that in eutopic endometrium. ③Expression of ERK1/2 mRNA:The eutopic endometrium expression of danazol dose group, the HuPoSan medium and the low dose groups were lower than those in the model group. The eutopic and eutopic endometrium expression of Danazol group and low/medium/high dose group were no statistical significan-ce between each two.The ectopic endometrium expression of danazol dose group, the Hu PoSan medium dose group were higher than those in the model group.④Expression of p38 MAPK mRNA:The eutopic endometrium expression of danazol dose group, the HuPoSan high dose group were lower than those in the model group. The eutopic endometrium expression of danazol dose group, the HuPoSan medium dose group were higher than the HuPoSan high/low dose group. The ectopic endometrium expression of danazol dose group, the HuPoSan high dose group were lower than those in the model group. The ectopic endome-trium expression of danazol dose group were lower than those in the HuPoSan low/medium/high dose groups. The ectopic endometrium expression of the HuPoSan high dose group were lower than those in medium dose group.⑤Expression of NF-κB mRNA:The eutopic endometrium expression of the HuPoSan high/low dose group was lower than those in the model group. The eutopic endometrium expression of the HuPoSan low dose group was lower than those in medium dose group. The eutopic endometrium expression of the HuPoSan low dose group was lower than those in the danazol group.There was no significant difference in the expression of ectopic endometrium between the disease model group and the danazol group, the HuPoSan low/medium/high dose groups. The ectopic endometrium expression of the danazol group was lower than those in the HuPoSan low/medium dose groups. The ectopic endometrium expression of the HuPoSan high/low groups was lower than those in the medium dose group.(4)Conclusion:The activation of HuPoSan Warming Kidney and Removing Blood Stasis can inhibit ERK1/2 and p38MAPK pathway in eutopic and ectopic endometrium. Because MAPK pathway is involved in nuclear transcription factor expression, so the HuPoSan may reduce the release of NF kappa B by inhibiting the activity of MAPK pathway. Then it reduce cytokines, chemotactic factor, adhesion molecules, growth factors, apoptosis related gene expression, regulating the immunity and inflammation, apoptosis and cell proliferation. differentiation, and to slow down the development of EMS.The subject about exploring the molecular mechanism of the Warming Kidney and Removing Blood Stasis method on inhibiting ectopic endometrial adhesion, invasion and angiogenesis, which will increase the new content for Chinese medicine in the treatment of EMS, and provide a new way for the study of its pathogenesis, and then provide more theoretical basis for its clinical treatment.2.Summary of Professor Wang Guohua’s experience:Based on the experimental research part that reveals the material basis and biology connotation of HuPoSan in the treatment of Ems model rat, further to summed up the clinical treatment of Ems experience. Understanding of the etiology and pathogenesis of EMS, except now recognized as "blood stasis of uterus, Chong and Ren", We should also pay attention to the cold and kidney. The treatment should be considered both, besides the promoting blood circulation and removing blood stasis, also pay attention to the treatment of warming kidney. So Yang is filled, the uterus, Chong and Ren can keep warm, then Qi and Blood can circulation, Yin Xie powder,cold junction to cancel. With a case, to analysis Professor Wang Guohua’s clinical treatment of Ems and prescription medication characteristics. |
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