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Research on Regulation Effect of Dissipating Phlegm and Removing Stasis Method on MMPs-TIMPs Expression Balance of Polycystic Ovary Rats

Author: LiYan
Tutor: ZhangQingWen
School: Chengdu University of Traditional Chinese Medicine
Course: Chinese Gynecology
Keywords: polycystic ovary syndrome DHEA matrix metalloproteinases tissue inhibitors of metalloproteinases sanbeidijiajian
CLC: R271.9
Type: PhD thesis
Year: 2013
Downloads: 11
Quote: 0
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Abstract


Objective:The dehydroepiandrosterone (DHEA) was used to induce the model of SD rats with polycystic ovary syndrome (PCOS), in order to research the matrix metalloproteinases of polycystic ovary rats and the imbalance of their tissue inhibitory factors (MMPs-TIMPs), then the Chinese medicine Sanbeidijiajian (Compound medicine of Rhizoma Sparganii and Fritillaria etc.) with dissipating phlegm and removing stasis efficacy was used to regulate the MMPs-TIMPs expression balance of polycystic ovary rats, so as to provide a scientific basis for the treatment of PCOS.Methods:1. The DHEA solution was administrated to the rats daily by subcutaneous injection (6mg/0.2ml/100g) for20consecutive days in order to induce the models of PCOS, and then the reproductive hormone T, LH, FSH values, blood glucose and insulin levels, ovarian histology indicators of both blank group and model group were determined by ELISA assay, the DEHA were verified to build PCOS models, next, the2-ΔΔCT values and the MMPs/TIMPs ratios of the ovarian MMP-2mRNA、MMP-9mRNA、 TIMP-1mRNA、TIMP-2mRNA were determined by QPCR assay, so as to study the expression balance of the of MMPs-TIMPs under PCOS condition. All data were collected and analyzed by using SPSS18.0statistical software. Measurement data were expressed as form of mean±standard deviation (x±s), and the data of both groups were compared independent samples Student’s t-test.2. After the application of DHEA for the formation of PCOS models, the rats were randomly divided into7groups including blank group, saline group, Diane group, metformin group, Chinese medicine Sanbeidijiajian high, middle, low dose groups and were orally administrated their corresponding medicines, and they were performed vaginal exfoliative cytology smear test daily, when50%of rats in one of model groups had their first normal ovulation cycles, all the animals were sacrificed. Then the reproductive hormone T, LH, FSH values, blood glucose and insulin levels, ovarian histology indicators of every group were determined by ELISA assay, and the2-ΔΔ CT values and the MMPs/TIMPs ratios of the ovarian MMP-2mRNA、 MMP-9mRNA、TIMP-1mRNA、TIMP-2mRNA were determined by QPCR assay, so as to compare different effects produced by the medicines in every group and explore the treatment mechanisms of these medicines to PCOS based on their regulations on the expression balance of MMPs-TIMPs, especially Chinese medicine Sanbeidijiajian. All data were collected and analyzed by using SPSS18.0statistical software. Measurement data were expressed as form of mean±standard deviation (x±s), the single-factor variance analysis was used for multi-group comparison, and the LSD-t test was used for the data comparisons between any two of all these groups.Results:DHEA-induced rats had significant high ovarian indexes, and ovarian morphologic changes such as decreased mature follicles, increased atretic follicles, increased follicular cystic dilatations, increased serum T levels and LH values, and higher fasting blood glucose, fasting insulin, insulin resistance indexes than those of blank group, as well as high MMPs/TIMPs ratio, which were basically in line with the pathological features of PCOS. The Chinese medicine Sanbeidijiajian with dissipating phlegm and removing stasis efficacy can effectively reduce the ovarian indexes of PCOS rats and improve their ovarian morphological changes, as well as decrease their serum T levels, fasting blood glucose, fasting insulin, insulin resistance indexes, decrease their abnormal MMP9/TIMP1ratio and MMP2/TIMP2ratio, thereby regulating the expression balance of MMPs-TIMPs to produce a comprehensive effect better than that of positive drug result Diane-35or metformin. This medicine has an excellent therapeutic effect for PCOS.Conclusion:DHEA can be used to induce the SD PCOS rats model; the PCOS rats have MMPs-TIMPs expression imbalance; the Chinese medicine Sanbeidijiajian with dissipating phlegm and removing stasis efficacy can effectively regulate the expression balance of MMPs-TIMPs in PCOS rats.

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