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김장흡,김진홍,류순원,차정호,류기성,김미란,권동진,임용택,이윤진,조현희 대한산부인과학회 2002 Obstetrics & Gynecology Science Vol.45 No.4
난포의 성장과 배란에 중추적 기능을 하는 뇌하수체 호르몬인 FSH는 G 단백 결합 수용체군에 속하는 FSHR과 결합하여 작용한다. 난소의 노화는 난포의 수와 기능의 감소로 인한 estrogen과 inhibin 생성 감소의 결과로 FSH 상승을 유발한다. FSH 증가에도 불구하고 난소의 반응이 감소되어 있고 estrogen과 inhibin 생성이 저하되는 것이 폐경의 주된 기전으로 추정되므로 난포내 FSHR mRNA 발현 변화의 규명은 폐경 기전을 밝히는데 반드시 필요하다. 저자는 노화 및 혈중 FSH 상승에 따른 FSHR mRNA의 발현 변화를 in situ hybridization 및 QC RT-PCR을 이용하여 연구하였다. 실험 대상은 정상 월경주기를 가진 40세 이하, 40-44세, 45-49세, 50-54세 여성 각각 10명과, 그리고 폐경 이후 10명, 총 50예의 신선 난소조직과 혈액을 채취하여 실시하였다. In situ hybridization은 cloning된 FSHR의 DNA를 이용하여 digoxigenin이 부착된 약 800 bp의 ssRNA probe를 제작하고 파라핀 포매된 절편에서 시행하였으며, 염색정도를 FSH의 영향을 받지 않는 일차 난포 (primary follicle)에서 0, 1+, 2+ 등급으로 비교하였다. QC RT-PCR은 primer (antisense: 5’-GGCCCTGCTCCTGGTCTCTTTG-3’, sense: 3’-AACAGCGGGAGTACCTTCGG-5’)를 제작하여 799 bp의 PCR 산물을 생성토록 하였고, 목적 유전자와 경쟁적으로 작용하는 149 bp가 결실된 DNA competitor를 site-directed mutagenesis를 이용하여 제작한 후 목적 유전자와 함께 PCR하여 RNA 발현양을 비교 정량하였다. 결과는 Pearson 상관분석과 independent t-test를 이용하여 통계처리 하였다. 난포 수의 변화는 연령 증가 (r=-0.934, P=0.01)와 FSH 상승 (r=-0.713, P<0.001)에 따라 역상관 관계로 감소하였다. In situ hybridization에서 FSHR mRNA 발현 정도는 40세 미만에서는 비슷하게 발현되었으나 40세 이후에는 연령 증가와 FSH 상승에 반비례하여 (r=-0.744, P<0.001 과 r=-0.771, P<0.001) 감소하는 소견을 보였으며, 폐경 후의 난소에서는 발현을 볼 수 없었다. QC RT-PCR로 측정한 FSHR mRNA 발현량은 연령군에 따라 40세 미만에서 840.00±516.29, 40-44세에서 240.00±154.91, 45-49세에서 40.00±21.90, 50-54세에서 6.06±4.13, 폐경 후 여성에서 0.48±0.00 fg이었으며, 연령 증가와 FSH 상승에 반비례하여 (r=-0.857, P<0.001과 r=-0.771, P<0.001) 발현량이 감소하였다. 본 연구 결과 40세 이후의 FSH 상승은 난소 호르몬 변화뿐만 아니라 FSHR mRNA의 감소와도 연관되어 있으며, 일차 난포에서 연령에 따른 FSHR mRNA 감소는 FSH와 상관없는 난포 노화 때문이라고 생각된다. 따라서 폐경의 진행은 40세 이후에 이미 시작되며 여러 요인의 복합적인 작용 결과이지만, 그 주된 기전은 FSHR 발현 저하와 이에 따른 성선자극호르몬에 대한 난소의 반응저하로 추정된다. 폐경의 보다 정확한 기전 및 영향 인자 규명을 위하여 본 연구를 기초로 향후 더 많은 연구가 필요하리라 사료된다. FSH is the central hormone for the regulation of ovarian function and acts by binding with specific receptor, FSHR, which is one of the G-protein coupled receptor family. The aging of ovary decreases the number and the activity of follicle, which results in the increase of FSH by reduction of inhibin and estrogen. The study on FSH level and FSHR mRNA expression in the ovary of perimenopausal women is the crucial step for the understanding of the menopausal mechanism. We studied FSHR mRNA expression of ovarian follicle by using in situ hybridization and QC RT-PCR. The fresh ovarian tissues and blood samples were obtained from premenopausal women in mid-follicular stage and postmenopausal women. The experimental samples were grouped as below 40 years old women, 40-44 years old ones, 45-49 years old ones, 50-54 years old ones, and postmenopausal women as negative control. To localize FSHR transcripts by in situ hybridization, we synthesized digoxigenin-labelled ssRNA probe (about 800 bp) and measured the degree of staining as 0, 1+, 2+ in the primary follicles which were independent to FSH effect. To do QC RT-PCR, we synthesized oligonucleotide primers (antisense: 5-GGCCCTGCTCCTGGTCTCTTTG-3, sense: 3-AACAGCGGGAGTACCTTCGG-5) to form the 799 bp sized PCR products. We also synthesized 149 bp deleted DNA competitor by site-directed mutagenesis and then calculated the relative amount of target FSHR mRNA by comparing with competitor after PCR. There were significant reverse relationships between follicular number and aging (r=-0.934, P=0.01), and FSH level (r=-0.713, P<0.001). The similar amount of FSHR mRNA was expressed in the group of below 40 years by in situ hybridization. In the groups of above 40 years, the FSHR mRNA expression decreased progressively according to aging (r=-0.744, P<0.001) and FSH level (r=-0.771, P<0.001). But we could not find FSHR mRNA expression in menopausal ovaries. The amount of follicular FSHR mRNA was measured as 840.00±516.29 for the below 40 years group, 240.00±154.91 for the 40-44 years group, 40.00±21.90 for the 45-49 years group, 6.06±4.13 for the 50-54 years group, and 0.48±0.00 fg in the postmenopausal ovary. The amount of FSHR mRNA decreased with ovarian aging (r=-0.857, P<0.001) and FSH level (r=-0.771, P<0.001). These results demonstrate that the gradual increase of FSH and the decrease of FSHR mRNA expression in older than 40 years women are related to the changes of sex hormones. However the gradual decrease of the FSHR mRNA expression in the primary follicle may be due to the follicular aging itself. Therefore the menopausal transition already starts at the beginning of 40 years and one of the major cause of the menopause may be the reduction of FSHR mRNA expression followed the decrease of ovarian response to gonadotropins. The further studies should be required to elucidate the underlying mechanism and the associated factors of menopause.
긴장성 요실금 여성에서 vaginal cone을 이용한 골반저 근육운동에 대한 평가
김진홍(Jin Hong Kim),류순원(Sun Won Yoo),길기철(Ki Cheol Kil),천연희(Yeon Hee Cheon),최형주(Hyoung Ju Choi),이지은(Ji Eun Lee),조현희(Hyun Hee Jo),나종구(Jong Gu Rha),김수평(Soo Pyung Kim) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.3
N/A Defective or inadequate pelvic floor function is important etiologic factor for urinary stress incontinence and uterine prolapse. Pelvic floor muscle exercise is the mainstream of the nonoperative treatment for female stress urinary incontinence. Especially Vaginal cones have been known as a simple and practical means of improving both pelvic floor muscle strength and genuine stress incontinence using biofeedback mechanism. Forty adult women suffering from mild and moderate stress incontincnce were instructed to exercise their pelvic muscles using vaginal cones. They retained cones of increasing weight in the vagina by contracting pelvic floor muscles for 15 minutes twice a day for 4 weeks. The comparison of pelvic floor muscle function before and after the exercise were assessed by the frequency of incontinence, vaginal digital palpation, vaginal pressure measurement and speculm lift test. The results were as follows ; 1. The frequency of incontinence was significantly reduced after the exercise using vaginal cone (P<0.001). 2. Vaginal digital palpation score (strength of vaginal levator muscle) was significantly increased after the exercise using vaginal cone (P<0.001). 3. The vaginal pressure by perineometer was significantly increased after the exercise using vaginal cones (P<0.001). 4. The vaginal lumen during contraction was significantly narrower after the exercise using vaginal cones (P<0.001). 5. The maximal cone weight which patients could hold over 1 minute was significantly increased after the exercise using vaginal cones (P<0.001).
폐경 전, 후 여성에 있어서 골밀도 예측을 위한 골대사 표지물질의 의의
이진우,김진홍,김미란,정기욱,류순원,권동진,임용택 대한폐경학회 1999 대한폐경학회지 Vol.5 No.1
"Recently, the biochemical markers for bone metabolism have been developed and are expected to reflect the minor change of bone turnover. Which bone tumover markers are predictive of bone mineral density (BMD) form the clinically important sites of lumbar spine and femoral neck according to the YSM remain unclear, and were the aim of this study. We compated bone formation markers: alkaline phosphatase (Alp), Osteocalcin and bone resorption markers: pyridinoline (Pyr), deoxypyridinoline (D-pyr) to see if they teflected the BMD according to the years since menopause (YSM), The subjects were divided into four groups based on their YSM. According to the groups, the mean levels of serum FSH in women 0-5YSM and 5-10YSM were significantly higher than the level in women -5-0YSM, and women 10-15YSM, also, was significantly higher than those of other groups were. And the mean level of serum E2 in women -5-OYSM was significantly higher than other groups. Pyr significantly increased compared in 0- 5, 5-10 and 10-15YSM compared with women -5-OYSM and D-pyr had an increased pattern without significance. There was no significance in Alp. and Osteocalcin according Groups. The BMD of femur neck were significantly increased in women 5-10YSM and 10-15YSM compared with women -5-OYSM and 0-5YSM. And the BMD of L2-4 and femur (Ward and Trochanter) were significantly decreased in women 0-5, 5-10 and 10-15YSM compared with women 5-OYSM. To evaluate the discrimination power of the three markers according to the groups. The Three markers and BMD were calculated according to the groups. Pyr had a strong negative correlation with BMD of L2-4 in women -5-0YSM, Osteocalcin had a negative correlation with BMD of L2-4 in women 5-10 and 10-15YSM. Pyr. in postmenopausal women significantly increased after menopause. BMD decreased later in femur neck than L2-4 and femur ward and trochanter. Marker for bone metabolism had a poup specificity, Pyr was good performance in women -5-OYSM and Osteocalcin was good performance in women 5-10 and 10-15YSM with BMD of L2-4 . "