Alleviating effect of Aucklandia lappa Decne on premenstrual syndrome

다국어 초록 (Multilingual Abstract)

Premenstrual syndrome (PMS) occurs in more than 80% of reproductive- age women and manifests as physical and physiological symptoms that develop during the luteal phase of menstrual cycle. The cause of PMS is not clear, but hypotheses include hormonal imbalances and neurotransmitter abnormalities. Prolactin hypersecretion is associated with decreasing LH, FSH, and progesterone levels and increasing estrogen.
We hypothesized that inhibiting prolactin hypersecretion would alleviate PMS symptoms. Regulation of prolactin expression in Aucklandia lappa Decne (AL) extract was measured in vitro using GH3 cells. To identify the prolactin inhibitor, AL roots were extracted and fractionated using 50% EtOH. Seventeen sub-fractions were separated from the ethyl acetate fraction of AL, which is effective in prolactin inhibition, and the final active components of syringin and (+)-Pinoresinol 8-O-β-D-glucopyranoside (PG) were separated from the ALE-15 sub-fraction. The mechanism by which AL, syringin, and PG inhibit prolactin was identified using dopamine receptors D2, ER-α, and PGR. In addition, the inhibition of inflammation by AL and syringin was confirmed by measuring NO products and infectious disease-related biomarkers in RAW 264.7 cells. The effects of AL on prolactin and hormone secretion, neurotransmitters, and uterine inflammation were also measured using a hyperprolactinemic rat model.
AL was confirmed to aid in hormone and neurotransmitter recovery by inhibiting inflammation and prolactin secretion in rats with hyperprolactinemia. AL significantly suppressed the expressions of inflammatory factors interleukin-6, tumor necrosis factor-α, and cyclooxygenase-2 in the uterine, and inhibited the secretion of prostaglandin E2 in the serum. Regulation of the estrogen receptor alpha expression of AL was confirmed in pituitary tissue, suggesting a function in hormone regulation.
In conclusion, AL is expected to alleviate uterine pain and mental symptoms caused by PMS based on its action restoring hormonal imbalance in hyperprolactinemia mice and alleviating inflammation of uterine tissue. We also confirmed that AL, syringin, and PG inhibit prolactin via mechanisms that regulate dopamine receptors, ER-α, and PGR. Therefore, the results of this study suggest that AL has the potential as a treatment for alleviating PMS symptoms in the human body.

목차 (Table of Contents)

• 1. Introduction 1
• 1.1 Premenstrual syndrome 1
• 1.2 Correlations between prolactin increase and other hormones in PMS 5
• 1.3 Inflammation in PMS 6
• 1.4 Hyperprolactinemia rat models 7
• 1.5 Aucklandia lappa Decne 7
• 2. Materials and methods 8
• 2.1 Chemicals and reagents 8
• 2.2 Preparation of AL extracts 8
• 2.3 Isolation and spectroscopic data 9
• 2.4 HPLC analysis 9
• 2.5 Cell lines and culture 10
• 2.6 Cell cytotoxicity assay 10
• 2.7 Cell proliferation assay 10
• 2.8 Nitric oxide (NO) assay 11
• 2.9 Prolactin inhibition assay 11
• 2.10 qRT-PCR analysis 12
• 2.11 Western blot analysis 15
• 2.12 Hyperprolactinemia rat models 15
• 2.13 Statistical analysis 16
• 3. Results 17
• 3.1 Evaluation of PMS treatment efficacy of AL in vitro 17
• 3.1.1 Inhibition effect of cell proliferation in AL extract and prefemin 17
• 3.1.2 Determination of E2 concentration 19
• 3.1.3 Prolactin inhibitory effect of AL extract in GH3 cells 21
• 3.1.4 Separation of active substances and of structure of AL 23
• 3.1.5 HPLC analysis 31
• 3.1.6 Prolactin inhibition by syringin and PG, an active component of AL 33
• 3.1.7 Mechanism of inhibiting prolactin secretion in GH3 cells of AL, syringin, and PG 35
• 3.1.8 Inhibitory mechanism of AL, syringin, and PG on PRL secretin in GH3 cells 38
• 3.1.9 Inhibitory effect of AL and syringin on NO production in RAW 264.7 cells 40
• 3.1.10 Effects of AL and syringin on pro-inflammatory cytokines and inflammatory mediators in mRNA level of RAW 264.7 cells 42
• 3.2 PMS alleviating effect of A. lappa in hyperprolactinemic rat models 44
• 3.2.1 Body weight by AL in hyperprolactinemic rat model 44
• 3.2.2 Organ weight change by AL in hyperprolactinemic rat model 46
• 3.2.3 Efficacy of AL on secretion of hormones in the hyperprolactinemia rat model 48
• 3.2.4 Recovery of neurotransmitter on AL in the serum of hyperprolactinemic rat 50
• 3.2.5 Effects of AL on inflammatory mediator secretion and uterine inflammation in hyperprolactinemia rats model 52
• 3.2.6 ER-α expression in the pituitary gland of hyperprolactinemic rats 54
• 4. Discussion 56
• 5. References 61
• 국문초록 70