Spinal Sigma-1 Receptor-mediated Dephosphorylation of Astrocytic Aromatase Plays a Key Role in Formalin-induced Inflammatory Nociception

Choi, Hoon-Seong,Lee, Mi-Ji,Choi, Sheu-Ran,Smeester, Branden A.,Beitz, Alvin J.,Lee, Jang-Hern Elsevier 2018 NEUROSCIENCE Vol.372 No.-

<P><B>Abstract</B></P> <P>Aromatase is a key enzyme responsible for the biosynthesis of estrogen from testosterone. Although recent evidence indicates that spinal cord aromatase participates in nociceptive processing, the mechanisms underlying its regulation and its involvement in nociception remain unclear. The present study focuses on the potential role of astrocyte aromatase in formalin-induced acute pain and begins to uncover one mechanism by which spinal aromatase activation is controlled. Following intraplantar formalin injection, nociceptive responses were quantified and immunohistochemistry/co-immunoprecipitation assays were used to investigate the changes in spinal Fos expression and the phospho-serine levels of spinal aromatase. Intrathecal (i.t.) injection of letrozole (an aromatase inhibitor) mitigated both the late phase formalin-induced nociceptive responses and formalin-induced spinal Fos expression. Furthermore, formalin-injected mice showed significantly reduced phospho-serine levels of aromatase, which is associated with the rapid activation of this enzyme. However, sigma-1 receptor inhibition with i.t. BD1047 blocked the dephosphorylation of aromatase and potentiated the pharmacological effect of letrozole on formalin-induced nociceptive responses. In addition, i.t. administration of a sub-effective dose of BD1047 potentiated the pharmacological effect of cyclosporin A (a calcineurin inhibitor) on both the formalin-induced reduction in phospho-serine levels of aromatase and nociceptive behavior. These results suggest that dephosphorylation is an important regulatory mechanism involved in the rapid activation of aromatase and that spinal sigma-1 receptors mediate this dephosphorylation of aromatase through an intrinsic calcineurin pathway.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Dephosphorylated aromatase in astrocyte is involved in the process of formalin-induced acute nociception. </LI> <LI> Activation of astrocyte sigma-1 receptors is associated with the dephosphorylation of aromatase. </LI> <LI> Calcineurin signaling pathway is involved in the sigma-1 receptor-mediated dephosphorylation of aromatase. </LI> </UL> </P>

자궁내막증에서 aromatase의 발현과 임상적 중등도와의 연관성

이일한 ( Il Han Lee ),김동호 ( Dong Ho Kim ),노지현 ( Ji Hyun Noh ),고재환 ( Jae Whoan Koh ),김용봉 ( Yong Bong Kim ) 대한산부인과학회 2010 Obstetrics & Gynecology Science Vol.53 No.4

Objective: Aromatase is the key enzyme for the conversion of C19 steroids into estrogen in certain human tissues. We studied to evaluate the aromatase expression in eutopic endometirum and endometriotic lesion and its relationship to clinical and laboratory parameters. Methods: The study included 78 cases of endometriotic lesion and 14 cases of eutopic endometrium and 30 cases of normal uterine endometrium obtained through laparoscopic surgery and curettage. The frozen tissue specimens were examined by immunohistochemistry using aromatase. Clinical symptoms, laboratory findings, and operative findings were analyzed and compared in according to aromatase expression. Results: We observed positive immunohistochemical expression for aromatase in endometriotic lesion from 46/78 patients (59.0%). Aromatase expression was elevated in comparison to eutopic endometrium (5/14 patients, P=0.032) and the difference was more pronounced when eutopic endometriums from patients with endometriosis were compared with those of healthy controls (2/30 patients, P<0.001). Aromatase-positive patients had more moderate-to-severe chronic pelvic pain, higher CA-125 level significantly. Also in operative findings, severe grade endometriosis, bilateral endometriomas, and associated leiomyoma and adenomyosis were more frequent in aromatase positive patients. High values of white blood cell count, erythrocyte sedimentation rate, CA 19-9 were more frequent in aromatase positive patients notwithstanding insignificant differences. Conclusion: Unopposed local biosynthesis of estrogens by increased expression of aromatase in eutopic endometrium and endometrial tissue could be involved in the development or maintenance of endometriosis and other uterine estrogen-triggered diseases. Our findings suggest increased expression of aromatase may be related with severity, activity, and chronic pelvic pain in patients with endometriosis.

DDT Reduced Testosterone and Aromatase Activity Via ER Receptor in Leyding Cell

Lee, Kyung Jin,Wui, Seong Uk,Heo, Jin,Kim, Sun Hee,Jeong, Ji Yeon,Lee, Jong-Bin 한국환경독성학회 2003 환경독성보건학회지 Vol.18 No.2

본 연구는 환경호르몬(endocrine disruptors)으로 분류되었으며, 제초제로 널리 사용되었던 Dichlorodiphenyltrichloroethane(DDT)가 설치류의 생식세포 중 Leydig 세포의 testosterone(T) 생성억제 및 그 관련 작용메카니즘을 규명코자 수행되었다. 먼저 흰쥐의 웅성 생식세포주인 R2C 세포에 T의 양 및 aromatase 활성도를 radio immunoassay(RIA)방법을 이용하여 측정하였다. 그 결과 R2C 세포에 황체형성호르몬(LH)를 처리하여 testosterone 생성을 증사시킨 후, DDT를 처리한 군들은 대조군에 비하여 농도 의존적으로 T의 양은 감소하였으며, aromatase 활성도는 증가하였다. 또한 DDT 자체만 처리한 군에서도 대조군에 비하여 testosterone의 생성이 감소하였다. 이러한 aromatase 활성 증가가 estradiol receplor(ER)와의 상호 관련성을 확인하기 위해 ER antagonist인 ICI 182.780를 처리한 후 T의 양 및 aromatase 활성도를 측정한 결과 DDT에 의해 증가된 aromatase 활성도가 ICI 182.780에 의해 다시 감소됨을 확인하였다. 또한 DDT에 의해 감소된 T의 양도 ICI 182.780에 의해 다시 회복되었다. In vivo 실험으로 흰쥐에 DDT를 직접 투여한 후 정소 내 성 호르몬들을 측정해 본 결과 T의 양은 유의성 있게 감소하였으며, estradiol(E2)의 양은 증가하였으며, aromatase 활성도도 감소하였다. 이러한 결과를 종합해 볼 때 DDT는 aromatase를 감소시키고, 이렇게 감소된 aromatase에 의해 testosterone 생성량을 억제하고, 이러한 DDT의 aromatase의 감소는 ER을 경유하는 것으로 추정할 수 있다. Dichlorodiphenyttrichloroethane (DDT), is a wide-spread environmental pollutant. In this study, we investigated the effect of DDT on testosterone production through aromatase and investigated its molecular mechanism in testicular leydig cell, R2C. We investigated that the effects of DDT on testosterone production and its effects on aromatase activity in R2C cell by radio immunoassay (RIA). As the results, the potent leydig call activator LH increased testosterone production compared to the control. DDT exposure significantly decreased testosterone production in R2C cell and DDT alone affected T reduction in a dose-dependent manner in R2C cell slightly. In addition, DDT was found to increase aromatase activity in R2C cell in a dose dependent manner. In order to assess whether the suppressive effects of DDT on LH-inducible testosterone production might be influenced by the ER, ICI 182.780, a pure antiestrogen, was used, and it was found that these inhibitory effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the suppressive effects of DDT. Furthermore, the inducible effects of DDT on aromatase might be influenced by the ER, ICI 182.780 was used, and it was found that these enhancing effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the inducible effects of DDT. Our results indicated that DDT inhibition of LH-inducible testosterone production in R2C is mediated through aromatase. However, the precise mechanisms by which DDT enhance in leydig cell remains unknown. The current study suggests the possibility that DDT might act as a modulator aromatase gene transcription.

The Timing of Aromatase Action for Sex Differentiation in the Nile Tilapia, Oreochromis niloticus

권준영,권혁추 한국발생생물학회 2006 발생과 생식 Vol.10 No.3

어류의 체내에서 성분화를 유도하는 물질이 성스테로이드호르몬(sex steroid hormone)이라는 사실이 잘 밝혀져 있으며, 성스테로이드 생합성 효소의 하나인 aromatase도 성분화에 직접적인 역할을 하는 것으로 알려져 있다. 본 연구에서는 유전적으로 암컷인 틸라피아 자어(larvae) 집단을 aromatase 저해제(aromatase inhibitor, AI)인 Fadrozole로 침지 처리하여 초기 발생단계 중 어느 시기에 aromatase가 Sex steroids are generally considered as natural sex inducers in fish, and aromatase (cytochrome P450 aromatase) that catalyzes androgens into estrogens in the steroidogenic pathway is also known to be involved in sex differentiation. The timing of aromatase action is, thus, of central importance in the study of fish sex differentiation. We treated sexually undifferentiated tilapia (Oreochromis niloticus) larvae with , a non-steroidal aromatase inhibitor (AI), by immersing the fish in a solution containing AI during the sex differentiation period to narrow down the critical period of aromatase action. Fish were treated once at 11 or 13 days post fertilization (dpf), or twice at 11 and 13 dpf. The concentrations of AI at each time of the treatment were 0 mg/L (control), 50 mg/L or 100 mg/L. Survival rate was not statistically associated with AI immersion treatment (p>0.25). However, sex ratio was significantly altered by the treatment, with higher concentration and double immersion being more effective in masculinizing genetic females (p<0.05). These results suggest that aromatase action for sex differentiation in this fish species would begin at least from 11 dpf which is much earlier than previously expected, and that only 3 hours of brief immersion in AI solution is powerful enough to alter genetically programed sex.

Effect of DDT on Testosterone Production by Modulator Aromatase (CYP 19) in R2C

( Kyung Jin Lee ),( Jong Bin Lee ),( Hye Gwang Jeong ) 한국환경생물학회 2003 환경생물 : 환경생물학회지 Vol.21 No.3

N/A Abstract - Various pesticides known or suspected to interfere with steroid hormone function were screened for effects in leydig cells on catalytic activity and mRNA expression of aromatase. Dichlorodiphenyftrichloroethane (DDT) is a widespread environmental pollutant. In this study, we investigated the effect of DDT on testos-terone production through aromatase activity and its molecular mechanism in testicular leydig cell, R2C by using radioimmunoassay (RIA). As the results, the potent leydig cell activator LH increased testosterone production compared to the control. DDT exposure significantly decreased testosterone production in R2C cell. In addition, DDT was found to increase aromatase gene expression and activity in R2C cell in a dose dependent manner. In order to assess whether the suppressive effects of DDT on LH-inducible testosterone (T) production might he influenced by the ER, ICI 182.780 was used, and it was found that these inhibitory effects of DDT were antagonized by ICI 182.780, implying that the estrogen receptor (ER) mediates the suppressive effects of DDT. Furthermore, the inducible effects of DDT on aromatase gene expression might he influenced by the ER, ICI 182.780 was used, and it was found that these enhancing effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the inducible effects of DDT. Our results indicated that DDT inhibition of luteinizing hormone (LH)-inducihle T production in R2C cell is mediated through aromatase. However, the precise mechanisms by which DDT enhance in R2C cell remains unknown. The current study suggests the possibility that DDT might act as a modulator aromatase gene transcription.

자궁내막증 환자의 자궁내막과 난소 자궁내막종에서 gonadotropin-releasing hormone agonist 치료 후 aromatase와 cyclooxygenase-2의 감소

김영아 ( Young Ah Kim ),김미란 ( Mi Ran Kim ),이재훈 ( Jae Hoon Lee ),안상태 ( Sang Tae Ahn ),김지연 ( Ji Yeon Kim ),황경주 ( Kyung Joo Hwang ),이응수 ( Eung Soo Lee ) 대한산부인과학회 2007 Obstetrics & Gynecology Science Vol.50 No.2

Objective: We investigated whether GnRH agonists reduce aromatase cytochrome P450 and cyclooxygenase-2 by direct action in the eutopic endometrium of endometriosis and ovarian endometrioma. Methods: Endometrial specimens and endometriotic tissues were obtained from infertility women undergoing laparoscopic surgery. Biopsy samples of the endometrium were obtained before and after GnRH agonist therapy. The stromal cells of eutopic endometrium in women with endometroisis and ovarian endometrioma were cultured in the presence of GnRH agonist (leuprolide acetate 0, 1, 5, and 10 uM) for 24 hours. The expression of aromatase cytochrome P450 and COX-2 was examined by Western blot. Results: Protein of aromatase cytochrome P450 and COX-2 were decreased in the eutopic endometrium of patients treated with GnRH agonist for 3 months. The stromal cells culture of endometrial explants and ovarian endometrioma with GnRH agonist reduced aromatase cytochrome P450 and COX-2. Phosphorylated ERK was decreased in endometriotic stromal cultures with GnRH agonist. Conclusion: These findings demonstrate that GnRH agonist not only promoted a hypoestrogenic state but also reduced aromatase cytochrome P450 and COX-2 by direct action on eutopic endometrium in patients with endometriosis and ovarian endometrioma.

해양심층수의 cytochrome P450 1A1, aromatase 및 MMP-9 활성 억제 효과

손윤희(Yun-Hee Shon),김미경(Mee-Kyung Kim),남경수(Kyung-Soo Nam) 한국생명과학회 2008 생명과학회지 Vol.18 No.4

동해 해양심층수의 유방암예방 효능과 전이에 미치는 영향을 알아보기 위해 cytochrome P450 1A1 활성과 aromatase 활성 및 유방암세포의 침윤성, 이와 관련된 MMP-9의 활성과 그 단백질 발현에 미치는 영향을 조사하였다. 해양심층수는 체내외의 여러 화학물질을 체내에서 활성화시켜 발암이나 돌연변이 등을 유발시키는 것으로 알려진 cytochrome P450 1A1을 경도의존적으로 저해시켰다. 또한 호르몬 의존성 유방암의 진행에 관여하는 aromatase의 활성도 경도의존적으로 저해시켰다(5.6∼51.9%). 해양심층수 처리에 의해 사람유방암세포인 MDA-MB-231 세포의 침윤성은 73.7∼29.4%로 감소하였으며, 세포의 침윤시 작용하는 단백질 분해 효소인 MMP-9의 활성과 단백질 발현도 경도의존적으로 억제되었다. 따라서 해양심층수는 유방암 예방과 전이관련의 더 많은 연구에 의해 유방암 예방과 전이 억제작용을 증명할 수 있을 것으로 보인다. Deep sea water from the East sea was tested for breast cancer chemoprevention and metastasis by measuring the activities of cytochrome P450 1A1 and aromatase, invasiveness, and activity and expression of matrix metalloproteinase (MMP)-9 in breast MDA-MB-231 cancer cell. The in vitro incubation of rat liver microsome with deep sea water (a hardness range of 100∼1,000) showed a hardness-dependent inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P450 1A1 activity. Deep sea water showed 27.1, 45.4 and 51.9% inhibition of microsomal aromatase activity at the hardness of 600, 800 and 1,000, respectively. In addition deep sea water inhibited not only the invasiveness of 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated MDA-MB-231 cells through matrigel-coated membrane in a hardness-dependent manner but also the activity and expression of MMP-9 in MDA-MB-231 cell.

Letrozole 및 Anastrozole에 의한 반구진 발진 1예

강혜련,김선,김현지,윤진,서장호,이주연 대한약물역학위해관리학회 2023 약물역학위해관리학회지 Vol.15 No.1

Aromatase inhibitors are widely used as a treatment for postmenopausal women with hormone receptor-positive breast cancer. They are classified as type 1 steroidal or type 2 nonsteroidal agents. Letrozole and anastrozole are the most frequently used type 2 nonsteroidal inhibitors that share common structure and bind reversibly to aromatase. Both are generally well tolerated; while type A adverse reactions such as hot flashes, vaginal dryness, musculoskeletal pain, and headache can occur, drug hypersensitivity reactions are very rare. Here, we present a case of a 55 years old woman who experienced maculopapular rash to letrozole after five days administration. Her symptoms resolved after chlorpheniramine and dexamethasone injection. Then, she took a single dose of letrozole and symptoms were aggravated. Sixteen days after her initial symptom to letrozole, she switched to anastrozole. After taking anastrozole for two days, she developed urticarial rash and stopped anastrozole arbitrarily. Her symptoms resolved after discontinuing anastrozole. Patients and physicians should be aware of potential cross reactivity to both aromatase inhibitors which may be due to the common structure in the aromatase inhibitors.

Cell Growth Inhibition and Gene Expression Regulation by (-)-Epigallocatechin-3-Gallate in Human Cervical Cancer Cells

Yanyan Qiao,Xiaolin Shi,Jinyan Cao,Liangqun Xie 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.9

EGCG [(-)-epigallocatechin-3-gallate] has shown its antitumor ability and perhaps a potential regimen for cancer patients. The goal of this study was to investigate the effect of EGCG on human papilloma virus (HPV) positive cervical cancer cell lines. EGCG inhibited the growth of CaSki (HPV16 positive) and HeLa (HPV18 positive) cells in a time- and concentration-dependent manner. Cell cycle arrest and apoptosis were observed in two cell lines after EGCG exposure. More importantly, we focused on EGCG regulation ability on pivotal genes involved in cervical cancer: viral oncogenes E6/E7, estrogen receptor (ER) and aromatase. Our results suggested that EGCG may be suitable for prevention and treatment of cervical cancer.

Effect of aromatase on sperm fertility and relative quantity of sperm aromatase protein in pig

Jong-nam Oh,Jae Yeon Hwang,Dong-Kyung Lee,Kwang-hwan Choi,Chang-Kyu Lee 한국발생생물학회 2014 한국발생생물학회 학술발표대회 Vol.2014 No.9

Aromatase is an enzyme that converts testosterone to estrogen. This enzyme, present in the sperm as well as various tissue and cells, has been considered to be related to the fertility of human and mouse sperm. Therefore, we examined effect of aromatase inhibitor on viability and fertility of sperm, and quantity of aromatase in sperm groups with different density in pig. To analyze the effect of aromatase on sperm viability, we treated aromatase inhibitor to the sperm with different concentrations (0, 10, 20, 50, 100, 200, 500 μM) at different time (0.5, 1, 2, 4, 8 hours). After the treatment, the sperm viability was calculated by hypo-osmotic swelling test. We selected 0, 50, 100 μM concentration during 0.5 hour as inhibitor treatment condition before in vitro fertilization. Next, we examined fertility and quantified aromatase protein in sperms with different density. In the first experiment, viability of sperm was decreased following the increasement of inhibitor concentration. The aromatase inhibited sperm showed lower penetration rate and cleavage rate than those of non-treated sperm. Concentration of 50 μM inhibitor had no significant effect on the sperm viability, but it significantly reduced sperm fertility. Second, sperms with low density showed higher penetration rate, but no significant difference between sperms with high density. In conclusion, aromatase is responsible for viability and fertility of porcine sperm similar to mouse and human, however, density of sperm has no correlation with quantity of aromatase protein.