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The Traditional Korean Herbal Medicine Ga-Gam-Nai-Go-Hyan

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The Traditional Korean Herbal Medicine Ga-Gam-Nai-Go-Hyan EXPERIMENTAL AND THERAPEUTIC MEDICINE 13: 1025-1031, 2017 The traditional Korean herbal medicine Ga-Gam-Nai-Go-Hyan suppresses testosterone-induced benign prostatic hyperplasia by regulating inflammatory responses and apoptosis SU-JIN SHIN1*, KWANG‑HO LEE2*, KYUNG‑SOOK CHUNG1, SE‑YUN CHEON1 and HYO‑JIN AN1 Departments of 1Pharmacology, and 2Acupuncture and Moxibustion Medicine, College of Korean Medicine, Sangji University, Wonju, Gangwon 26339, Republic of Korea Received September 14, 2015; Accepted October 28, 2016 DOI: 10.3892/etm.2017.4088 Abstract. Benign prostatic hyperplasia (BPH) is a patho- Introduction logical condition that affects the majority of men above the age of 50 years. Pharmacological agents are typically used to treat Benign prostatic hyperplasia (BPH) is one of the most BPH; however, there are currently no pharmacological agents common chronic diseases in men (1). A previous epidemio- that are able to completely cure BPH without causing adverse logical study has determined that >50% of men over the age of side effects. As a result of these side effects, there is a great 50 years exhibit symptoms of BPH, such as urinary urgency interest in developing effective herbal medicines that are able and retention, and the incidence rate gradually increases to inhibit the progression of BPH and are safe for long-term use. with age (2). Aging and androgens are known to be the two Ga-Gam-Nai-Go-Hyan (GGN) is a traditional Korean herbal main factors associated with the development of BPH, and medicine that has been widely used to treat BPH; however, it has previously been reported that inflammation may be no biological studies have been performed to elucidate the another key factor in prostatic enlargement (3,4). In addition, efficacy of GGN. The aim of the present study was to evaluate repeated tissue damage due to chronic inflammation may the efficacy of GGN as a treatment for BPH. GGN admin- provoke compensatory cellular proliferation, which increases istration was demonstrated to significantly decrease prostate the risk of hyperplastic growth (3,5). Chronic inflammation weight (P<0.001), the relative prostate weight ratio (P<0.001) is able to induce proliferation in prostate tissue by affecting and the ratio of prostate weight to body weight (P<0.001). apoptotic protein expression. Cyclooxygenase (COX)‑2 In addition, GGN treatment was revealed to suppress testos- inhibition is able to significantly increase apoptotic activity terone and dihydrotestosterone serum levels (P<0.001) and the in prostate cells via directly upregulating B-cell lymphoma growth of prostatic tissue. GGN also decreased the levels of (Bcl)‑2 expression (6). Cell growth in the normal prostate is the two inflammatory proteins (P<0.05), inducible nitric oxide regulated via a delicate balance of cell death and prolifera- synthase and cyclooxygenase‑2, decreased the levels of the two tion; therefore, disruption of the molecular mechanisms that apoptotic suppressors (P<0.05) B-cell lymphoma (Bcl)-2 and regulate these processes may lead to a state of epithelial and Bcl‑xL and increased the levels of the pro‑apoptotic factors stromal hyperplasia (7,8). (P<0.05) Bcl-2-associated X protein, caspase-3, caspase-8, At present, therapy for BPH is largely based on the use Fas, Fas ligand and Fas-associated protein with death domain. of α1-andrenergic receptor blockers and 5α-reductase inhibi- The results of the present study suggested that GGN may have tors, which relax prostatic smooth muscle and reduce prostatic suppressive effects on the development of BPH and therefore volume, respectively (9,10). Data from a previous systematic have the potential to be used for treating BPH. review indicates improved urine flow, nocturia and quality of life resulting from these treatments when used in combination or as monotherapies (11). Recently, the use of phosphodies- terase type 5 (PDE5) inhibitors was also recognized as an effective treatment for BPH (12). These pharmacological Correspondence to: Professor Hyo-Jin An, Department of agents are effective; however, they induce undesirable adverse Pharmacology, College of Korean Medicine, Sangji University, effects, including blood loss, urinary incontinence, infec- 83 Sangjidae Road, Wonju, Gangwon 26339, Republic of Korea tion, sexual dysfunction and morphological changes in the E-mail: [email protected] prostate (13,14). Therefore, it is necessary to identify novel *Contributed equally effective herbal products for the treatment of BPH that are also safe for long-term use. Key words: benign prostatic hyperplasia, Ga-Gam-Nai-Go-Hyan, Ga-Gam-Nai-Go-Hyan (GGN) is an oriental herbal inflammation, apoptosis, B-cell lymphoma 2 blend that has been used for a long time in Korea (15). It is composed of nine herbs: Morinda officinalis Haw, Cistanche salsa, Cornus officinalis, Cuscuta japonica 1026 SHIN et al: EFFECT OF GA‑GAM‑NAI‑GO‑HYAN ON BENIGN PROSTATIC HYPERPLASIA Chois, Psoralea corylifolia L, Dendrobium nobile Lindley, Table I. Recipe of Ga-Gam-Nai-Go-Hyan formulation used. Trigonella foenumgraecum L, Foeniculum vulgare Mill and Aconitum carmichaeli Debeaux (Table I). It has previ- Species Parts used Weight (g) ously been reported that GGN is effective in the treatment of pallor, dizziness, chronic prostatitis, impotence and Morinda officinalis Haw Roots 120 BPH (15). In addition, GGN has been used to treat patients Cistanche salsa Context stem 120 with genital herpes, which may support its use in the treat- Cornu sofficinalis Fruit 120 ment of GGN, as Trichomonas vaginalis has been detected Cuscuta japonica Chois Seed 120 in the urine of patients with BPH, which may indicate an Psoralea corylifolia L Seed 100 association between infections of the genital system and Dendrobium nobile Lindley Above ground 80 BPH (16). A Korean medicine book entitled The Treasured Trigonella foenumgraecum L. Seed 80 Mirror of Eastern Medicine reported that the primary Foeniculum vulgare Mill. Fruit 40 therapeutic facets of herbs used in GGN are similar to those Aconitum carmichaeli Debeaux Roots 20 of therapeutic agents used in the treatment of BPH (17). Certain major active components of GGN, including quer- cetin, kaempferol, coumarins and lignin glycosides, have been previously reported to exhibit anti‑inflammatory and anti‑oxidative qualities (18-21). Furthermore, a recent study The combination of herbs was extracted with 50% (v/v) by our group revealed that Cistanche salsa extract elicits an ethanol‑water at 60˚C for 8 h. The extracts were then filtered anti-proliferative effect on the prostate tissue of rats with through 15-µm cartridge paper and ethanol was removed via BPH (22). Although studies on the physiological functions of vacuum rotary evaporation (Eyela; Tokyo Rikakikai Co., the major active components of GGN have been performed, Ltd., Tokyo Japan). The concentrates were freeze-dried and the molecular mechanism(s) underlying the effect of GGN the yield was 12%. Powders were dissolved in distilled water on BPH have not yet been investigated; therefore, the aim of prior to experiments and residual powders were stored at the present study was to assess the anti-proliferative effects ‑20˚C. of GGN in a testosterone-induced rat model of BPH, and to demonstrate that it functions through regulation of the Animals. Ten-week-old male Wistar rats (n=24; weight, inflammatory response and apoptotic protein expression. 200±20 g) were purchased from Daehan Biolink Co., Ltd., (Republic of Korea). Rats were housed under constant condi- Materials and methods tions (temperature, 22±2˚C; humidity, 55±9%; 12 h light/dark cycle; ad libitum access to food and water) in accordance Materials and reagents. All herbs used to prepare GGN with the Guide for the Care and Use of Laboratory Animals. were purchased from Omniherb (Dong Woo Dang Pharmacy All procedures were approved by the Ethics Committee for Co. Ltd., Yeongcheon, Korea). Finasteride was obtained Animal Care and the Use of Laboratory Animals at Sang-ji from Merck & Co., Inc. (Whitehouse Station, NJ, USA). University (Wonju, Korea; approval documents, 2013-03;) Antibodies against inducible nitric oxide synthase (iNOS; and conducted in accordance with University guidelines. Rats M‑19; sc‑650), COX‑2 (C‑20; sc‑1745), procaspase‑3 (E‑8; were randomly distributed into four groups (n=6 in each); the sc-7272), procaspase-8 (C-20; sc-6136), B-cell lymphoma-2 sham-operated group (Con; administered with 200 µl distilled (Bcl‑2; C‑2; sc‑7382), Bcl‑extra large (Bcl‑xL; H‑5; sc‑8392), water orally), the BPH model group (BPH), the BPH-induced Bcl‑2‑associated X protein (Bax; B‑9; sc‑7480), p53 (FL‑393; group administrated with finasteride (Fina; 5 mg/kg/day; p.o. sc-6243), Fas (A-20; sc-1023), Fas ligand (Fas-L; C-178; Merck & Co., Inc.) and the BPH-induced group administrated sc-6237) and β-actin (ACTBD11B7; sc-81178) were purchased with GGN (GGN; 100 mg/kg/day; p.o.). BPH was induced in from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). An rats via castration and subsequent subcutaneous injections antibody against Fas-associated protein with death domain of 100 µl testosterone propionate (Waco Pure Chemical (FADD; ab24533) was purchased from Abcam (Cambridge, Industries, Ltd., Osaka, Japan; 10 mg/kg/day), as previously UK). The horeseadish peroxidase‑conjugated secondary reported (23). At the end of the four-week period, body weight antibodies (goat anti-rabbit, 111-035-003; rabbit anti-mouse, was recorded and rats were fasted for 12 h. The following 315-035-003; and donkey anti-goat, 705-035-003) were day, all rats were sacrificed by cervical dislocation following purchased from Jackson ImmunoResearch Laboratories, Inc. anesthetization with Zoletil 50 (20 mg/kg intraperitoneally; (West Grove, PA, USA). All other reagents were purchased Virbac, Carros, France), and blood samples were obtained via from Sigma-Aldrich (Merck Millipore, Darmstadt, Germany). cardiac puncture.
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