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???雅녶쑉翁ラ쳵罌?訝?쉪?방츏????잒꺗?╈?
??⒵낏? 10-0997215
Eur J Pediatr (2002) 161: 406?407
DOI 10.1007/s00431-002-0955-3
RESEARCH LETTER
Kanghyun Leem 횈 Sun-Young Park 횈 Dong-Hwan Lee ?쒎ㄷ庸좂뵟?100關m, 24弱??????╈?Hocheol Kim
?域꿨컦?燁??恙?(Eur J Pediatr (2002) 161: 406-407)
Lovastatin increases longitudinal bone growth and bone morphogenetic
protein-2 levels in the growth plate of Sprague-Dawley rats
Received: 9 July 2001 / Accepted: 25 February 2002 / Published online: 30 April 2002
? Springer-Verlag 2002
Lovastatin increases longitudinal bone growth and bone 5 mg/kg lovastatin for every 5 consecutive days
morphogenetic protein-2 levels in the growth plate of according to [5], while those of the control group were
Sprague-Dawley rats and may have implications for the given equivalent injections of normal saline. The oral
treatment of children. intakes and body weights of both groups were measured
but did not show any statistical di?erence (data not
Lovastatin is a drug widely used to reduce cholesterol shown). On the 3rd and 5th days after administration,
levels in blood. It is known to act via the inhibition of 3- all animals were injected intraperitoneally with
hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) 10 mg/kg tetracycline for measurement of bone growth.
reductase. Recently, Mundy et al. [5] showed that lo- On the 6th day, animals of each group were totally
vastatin increases bone morphogenetic protein-2 (BMP- anaesthetised with ether and sacri?ced 1 h after an
2) levels in human MG-63 osteoblasts, and suggested intraperitoneal injection of 5-bromo-2占?deoxyuridine
that it could be used to treat the loss of bone mass in (BrdU, 30 mg/kg) to determine the incorporation into
rodents. There was, however, no study on longitudinal newly synthesised DNA. The proximal tibia were
bone growth. Keller et al. [3] reported that the longitu- excised, ?xed for 24 h in 4% paraformaldehyde, and
dinal growth rate of the lower leg was reduced in pre- decalci?ed by immersion in 10% EDTA for 24 h. Each
mature infants. Accordingly, we set out to investigate sample was sectioned longitudinally (section width
whether lovastatin increases longitudinal bone growth in 40 lm). Bone growth was calculated by measuring the
the growth plate of the adolescent male rat. gap between successive bands formed by the injected
tetracycline using a ?uorescence microscope [2]. Ex-
Male Sprague-Dawley rats of 3 weeks of age were pressed BMP-2 and incorporated BrdU were detected by
used in the experiment. The experimental procedures avidin-biotin-peroxidase complex-linked immunohisto-
were performed in accordance with the animal care chemistry [1,4]. The proliferating ratio of chondrocytes
guidelines of the NIH and the Korean Academy of in growth plates for 1 h was measured by calculating
Medical Sciences. Animals were housed under con- labelling indices of BrdU positive chondrocytes [6].
trolled temperature (20짹2?C) and lighting (07:00?
19:00 h) conditions, with food and water made available The longitudinal bone growth rate for 48 h in normal
ad libitum. Rats were divided into two groups (n=5 in adolescent rats was 489.0짹70.1 lm and lovastatin was
each group): the control group and the lovastatin-ad- shown to promote bone growth, increasing the rate to
ministrated group. Animals of the lovastatin-adminis- 780.5짹72.8 lm (Fig. 1A) (P<0.05). The labelling index
trated group were injected intraperitoneally with for proliferating chondrocytes of the lovastatin-admin-
istrated group was 32.5짹5.7% compared to the control
K. Leem group (19.0짹7.8%, P<0.05). Both the number of
Department of Herbal Pharmacology, BMP-2-positive cells and the intensity of BMP-2 ex-
College of Pharmacy, Woosuk University, pression in proliferating, maturing, and hypertrophic
Chonbuk, South Korea chondrocytes of growth plates were increased in the lo-
vastatin-administrated group (Fig. 1C) compared with
S-Y. Park 횈 D-H. Lee 횈 H. Kim (&) the control group (Fig. 1B).
Department of Herbal Pharmacology,
Graduate School of East-West Medical Science, The increase in body length is mostly due to the
Kyung Hee University, 1 Hoegi-Dong, growth of longitudinal bones and their growth is a re-
Dongdaemoon-Ku, Seoul 130-701, South Korea ?ection of the proliferation and di?erentiation of
E-mail: heavenok@dreamwiz.com chondrocytes in the growth plates. Interestingly, lo-
Tel.: +82-2-9610419 vastatin increased longitudinal bone growth signi?cantly
Fax: +82-2-9640325
?쒎쑉???塋η쉪雅??繇??訝??雅녶끀鵝쒐뵪?╈?
雅??繇?????, 2008, 2017
JOURNAL OF MEDICINAL FOOD PHYTOTHERAPY RESEARCH Molecules 2013, 18, 13271-13282; doi:10.3390/molecules181113271 molecules
Phytother. Res. 26: 1771?1778 (2012)
J Med Food 13 (6) 2010, 1376?1384 Published online 3 March 2012 in Wiley Online Library Article
(wileyonlinelibrary.com) DOI: 10.1002/ptr.4642 Effects of Phlomis umbrosa Root on Longitudinal
# Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition Bone Growth Rate in Adolescent Female Rats
DOI: 10.1089=jmf.2010.1007 Skeletal Growth and IGF Levels in Rats after
HT042 Treatment Donghun Lee, Young-Sik Kim, Jungbin Song, Hyun Soo Kim, Hyun Jung Lee, Hailing Guo and
OPEN ACCESS Hocheol Kim *
The Herbal Formula HT042 Induces Longitudinal Bone Growth molecules Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, Seoul 130-701, Korea;
in Adolescent Female Rats ISSN 1420-3049 allstart2925@naver.com (D.L.); yjbsik@naver.com (Y.-S.K.); jbsong0527@gmail.com (J.S.);
Mi-Yeon Kim,1,2 Ji Young Kim,1 Dongwook Lim,1 Donghun Lee,2 Yoonjung Kim,2 www.mdpi.com/journal/molecules lovelyradix@hanmail.net (H.S.K.); lhjung91@hanmail.net (H.J.L.); guohailing1026@gmail.com (H.G.)
Mi-Yeon Kim,1,2 Youngmi Park,1 Naba Raj Pandit,1 Jiyoung Kim,1 Mikyung Song,1 Gyu Tae Chang,3 Ho-Young Choi1 and Hocheol Kim1,2* * Correspondence: hckim@khu.ac.kr; Tel.: +82-2-961-0419; Fax: +82-2-964-0325
Juyeon Park,2 Ho-Young Choi,1 and Hocheol Kim1,2 Communication
1Department of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University, Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Academic Editor: Derek J. McPhee
1Department of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University; Republic of Korea Effect of HT042, Herbal Formula, on Longitudinal Bone Received: 2 March 2016; Accepted: 31 March 2016; Published: 7 April 2016
and 2Korea Institute of Science and Technology for Eastern Medicine, NeuMed Inc., Seoul, Republic of Korea 2Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NeuMed Inc., Seoul 130-701, Republic of Korea Growth in Spontaneous Dwarf Rats
3Department of Pediatrics, College of Oriental Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea Abstract: This study aimed to investigate the effects of Phlomis umbrosa root on bone growth and
ABSTRACT The effect of HT042, a blend of three herbal extracts, on longitudinal bone growth was investigated in short- growth mediators in rats. Female adolescent rats were administered P. umbrosa extract, recombinant
and long-term rat models. In the short-term model, we divided female Sprague-Dawley rats (3 weeks old) into six groups, HT042, a new herbal prescription consisting of Astragalus membranaceus, Phlomis umbrosa and Eleutherococcus Ji Young Kim 1, MiKyung Song 1, Donghun Lee 1, Jungbin Song 1, Sang Woug Park 2, human growth hormone or vehicle for 10 days. Tetracycline was injected intraperitoneally to produce
according to treatment: vehicle, HT042 (100 mg=kg), Phlomis umbrosa (100 mg=kg), Astragalus membranaceus (100 mg=kg), senticosus, is used in traditional Korean medicine to stimulate growth in children. This study was conducted to inves- Juyeon Park 2, Seungjoon Park 3, Ho-Young Choi 1 and Hocheol Kim 1,* a glowing ?uorescence band on the newly formed bone on day 8, and 5-bromo-21-deoxyuridine
and Eleutherococcus senticosus (100 mg=kg) were administered twice daily, and recombinant human growth hormone (rhGH) tigate the effects of HT042 on skeletal growth, insulin-like growth factor-1 (IGF-1) and insulin-like growth factor was injected to label proliferating chondrocytes on days 8?10. To assess possible endocrine or
(1 IU) was subcutaneously injected once daily. Treatments were maintained for 4 days in each case. On day 3, tetracycline binding protein-3 (IGFBP-3) levels, and oestrogenic activity in female rats. Female Sprague?Dawley rats were 1 Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, autocrine/paracrine mechanisms, we evaluated insulin-like growth factor-1 (IGF-1), insulin-like
(20 mg=kg) was injected intraperitoneally (20 mg=kg) to form the ?uorescent band on the growth plates. On days 2?4, 5- divided into control, recombinant human growth hormone (rhGH; 20 mg/kg/day), and HT042 (100 mg/kg/day) growth factor binding protein-3 (IGFBP-3) or bone morphogenetic protein-2 (BMP-2) in response
bromo-20-deoxyuridine (BrdU) (50 mg=kg) was injected intraperitoneally to label proliferating cells. On day 5, the tibias were groups and treated for 3 weeks. Axial skeletal growth, femur length, and growth plate length were measured every Seoul 130-701, Korea to P. umbrosa administration in either growth plate or serum. Oral administration of P. umbrosa
dissected and ?xed in 30% sucrose. Dehydrated bone was sectioned at a thickness of 40 mm and observed. The bone growth in 3 weeks. The serum IGF-1 and IGFBP-3 levels were analysed. Moreover, the oestrogenic activity of the herbal 2 Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NeuMed Inc., signi?cantly increased longitudinal bone growth rate, height of hypertrophic zone and chondrocyte
groups administered HT042 and rhGH was signi?cantly increased to 433.50 짹 21.61 and 434.49 짹 15.21 mm=day, respectively, extracts in the immature and ovariectomized rats was tested. The nose?anus, nose?tail, femur and growth-plate proliferation of the proximal tibial growth plate. P. umbrosa also increased serum IGFBP-3 levels and
from 410.03 짹 17.4 mm=day (control). The height of the growth plates in the HT042 and rhGH groups was also signi?cantly lengths were increased signi?cantly in the HT042 group. Both IGF-1 and IGFBP-3 were highly expressed in the Seoul 130-701, Korea upregulated the expressions of IGF-1 and BMP-2 in growth plate. In conclusion, P. umbrosa increases
increased to 556.5 짹 21.1 and 544.2 짹 21.1 mm (P < .05), respectively, from 518.1 짹 4.1 mm (normal). The number of BrdU- hypertrophic zone of the growth plate. The serum IGF-1 levels were increased. Moreover, HT042 had no utero- 3 Department of Pharmacology, School of Medicine, Kyung Hee University, Seoul 130-701, Korea longitudinal bone growth rate by stimulating proliferation and hypertrophy of chondrocyte with the
positive cells in chondrocytes of the HT042 and rhGH groups was increased to 389 짹 36 and 627 짹 39 cells=mm2 (P < .001), trophic effects in the rats. Consequently, HT042 promoted longitudinal bone growth by stimulating cell proliferation increment of circulating IGFBP-3. Regarding the immunohistochemical study, the effect of P. umbrosa
respectively, from 264 짹 17 cells=mm2 (control). Insulin-like growth factor-1 and bone morphogenetic protein-2 in the HT042 in the epiphyseal plate and inducing the expression of IGF-1 without an oestrogenic response. HT042 may be helpful * Author to whom correspondence should be addressed; E-Mail: hckim@khu.ac.kr; may also be attributable to upregulation of local IGF-1 and BMP-2 expressions in the growth plate,
group were highly expressed in the growth plate. In the long-term rat model, the body weight, nose?tail length, and nose?anus in stimulating growth in children with short stature. Copyright ??2012 John Wiley & Sons, Ltd. Tel.: +82-2-961-0419; Fax: +82-2-964-0325. which can be considered as a GH dependent autocrine/paracrine pathway.
length were measured by microknemometry for 4 weeks. The body weight of the rhGH group was signi?cantly increased. The
nose?anus length of the HT042 and rhGH groups was signi?cantly greater at 18.5 짹 0.3 and 18.7 짹 0.3 cm compared to Keywords: HT042; herbal extract; skeletal growth; IGF-1; uterotrophic assay. Keywords: Phlomis umbrosa; bone growth rate; IGF-1; growth plate
18.2 짹 0.2 cm (control).
INTRODUCTION decreases protein concentrations in urine. Moreover, the Received: 23 September 2013; in revised form: 15 October 2013 / Accepted: 16 October 2013 /
KEY WORDS: ? Astragalus membranaceus ? 5-bromo-20-deoxyuridine ? chondrocyte ? Eleutherococcus senticosus administration of GH is limited by its poor absorption Published: 28 October 2013
? growth plate ? HT042 ? longitudinal bone growth ? Phlomis umbrosa ? recombinant human growth hormone It is widely recognized that the growth of children is a from the gastrointestinal tract. Regarding the economics
common marker of their health (Kurokawa et al., of the GH treatment, biosynthetic GH is expensive and
INTRODUCTION Angelica sinensis, Poria cocos, and Eleutherococcus senti- 2008) and that growth is regulated by the interaction in addition the short- and long-term bene?ts for the indi- Abstract: HT042 is a new herbal prescription consisting of Astragalus membranaceus, 1. Introduction
of genetic and acquired factors with environmental vidual are uncertain (Bryant et al., 2007). Phlomis umbrosa and Eleutherococcus senticosus, which are used in Korean medicine to
G rowth retardation in children can be caused by cosus. Furthermore, some herbs were reported to facilitate in?uences (Weedon et al., 2007), especially socioeco- stimulate growth in children. We investigated the effects of HT042 on the body weight, Short stature is de?ned as the height of an individual more than two standard deviation score
genetic, nutritional, metabolic, and=or endocrine fac- nomic conditions and nutritional status (Steckel, 2008). During bone growth, several indices and hormone (e.g. longitudinal bone growth, and bone length in spontaneous dwarf rats (SDR). Male and (SDS) below the average height for an age, sex, and population group [1]. Short children might have
tors. Malnutrition, metabolic acidosis, and growth hormone bone growth. A. membranaceus was reported to accelerate Growth retardation in children has a multifactorial ori- IGF-1, insulin-like growth factor binding protein-3 female SDRs were divided into three groups: the control group (DW, 10 mL/kg/day), the a high frequency of psychosocial problems such as lower self-esteem, social immaturity, or being
(GH) de?ciencies are conditions that can interfere with a gin. Factors that contribute to growth retardation include (IGFBP-3)) levels change. IGF-1 plays essential roles in recombinant human GH group (rhGH; 500 關g/kg/day), and the HT042 (100 mg/kg/day) bullied [2?5]. People with shorter height are reported to have a lower health-related quality of life
child?셲 ability to reach normal stature.1 GH replacement the proliferation and promotion of bone marrow stromal protein?calorie malnutrition, metabolic acidosis and pe- growth as a growth-promoting polypeptide. In serum, most group. Each group received the respective treatments for 10 days. Body weight was even if they do not ?t into the de?nition of short stature [6]. People with shorter height also have
therapy is often used to treat children with short stature, but cells2 and to induce GH in pituitary cell culture.3 E. senti- ripheral resistance to growth hormone (GH) in the target IGF-1 (70?80%) forms a 150-kD complex by binding measured on day 10 of treatment. On day 8, tetracycline (20 mg/kg) was injected higher morbidity of coronary heart diseases, perhaps because of the smaller diameter of their blood
many investigators have examined the possible ef?cacy of organs, as well as anaemia, renal osteodystrophy and with IGFBP-3. IGFBP-3 acts as a regulator of growth- intraperitoneally into all individuals to form a fluorescent band on the newly synthesized vessels [7,8].
traditional medicinal herbs. For thousands of years, tradi- cosus was reported to promote the longitudinal bone growth uremia itself altering the growth hormone/insulin-like dependent IGF-1 signalling through the enhancement of bone. On day 10, femur and tibia lengths were measured using PIXImus. Body weight,
tional medicinal herbs had been used to treat children with in adolescent male rats4 and to increase bone mineral density growth factor-1 (GH/IGF-1) axis (Dunkel, 2009). IGF-1 stability in serum or repression of IGF-1 binding to longitudinal bone growth, and bone length were not affected in the HT042 group. In Short stature caused by certain diseases, including growth hormone (GH) de?ciency, accounts
growth retardation, and the results have been recorded in the of the mouse tibia.5 the IGF-1 receptor (Binoux and Hossenlopp, 1988; Bar contrast, the rhGH group showed significantly increased body weight, longitudinal bone for 20% of all short children, but the remaining 80% are not determined, so-called idiopathic short
traditional Korean medicinal book, Dongeuibogam. Various The current methods for increasing ?nal height include et al., 1990). In the liver, circulating IGF-1 and IGFBP-3 growth, and bone length. In conclusion, HT042 does not act through a GH-like effect to stature (ISS) [1]. In case of GH de?ciency, average increase in ?nal height attributable to GH therapy is
herbal medicines for children had been recommended such Bone growth is a result of endochondral cell proliferation GH treatment alone or together with the gonadotropin- are synthesized by hepatocytes in a GH-dependent manner promote longitudinal bone growth. about 30 cm compared with predicted adult height [9,10]. In the case of ISS, the US FDA approved GH
as, among others, Panax ginseng, Astragalus membrana- releasing hormone (GnRH) analogue treatment (Richman et al., 2001; Roith et al., 2001). In addition, therapy in ISS children shorter than 쨈2.25 SDS in 2003 based on the evidence derived from two clinical
ceus, Atractylodes macrocephala, Glycyrrhiza glabra, in growth plates and conversion of chondrocytes into new (Leschek et al., 2004). However, excessive GH treatment IGF-1 is expressed in all non-hepatic tissues, where it acts Keywords: HT042; herbal extract; longitudinal bone growth; spontaneous dwarf rat studies [11,12], nevertheless, the average increase in ?nal height attributable to GH therapy in children
bone.6 To track these cells, cell labeling techniques using may hasten growth velocity and rapid progression in a paracrine/autocrine fashion (Yakar et al., 2002). with ISS is just 3.5?7.5 cm (4?7 years) [11,13?16]. In this case of ISS, the effect remains controversial
Manuscript received 18 January 2010. Revision accepted 20 July 2010. 5-bromo-20-deoxyuridine (BrdU) were developed. BrdU is a through puberty, and the combination of GH and GnRH because the estimated cost of GH therapy for adult height gain is about 10,000?20,000 dollars/cm [1,15].
agonists often decreases bone mineral density (BMD; In addition, oestrogens also play a pivotal role in
Address correspondence to: Hocheol Kim, Department of Herbal Pharmacology, College thymidine analog that incorporates into DNA of dividing Yanovski et al., 2003). In addition, GH administration pre- the regulation of normal skeletal growth and maturation
of Oriental Medicine, Kyung Hee University, 1 Hoegidong, Dongdaemoongu, Seoul 130- cells during the S-phase of the cell cycle.7 vents protein degradation in many diseases. Long-term in both boys and girls (Cutler, 1997). In both genders,
701, Republic of Korea, E-mail: hckim@khu.ac.kr treatment with GH increases lean body weight, muscle oestrogen de?ciency leads to the absence of a pubertal
For this bone growth study based on medicinal herbs we and lipid weights. Short-term treatment with GH growth spurt. When oestrogenic materials enter the
increases plasma insulin and IGF-1 concentrations and body via the intake of phytoestrogen and endocrine dis-
selected three herbs?the root of A. membranaceus, the stem ruptors, sexual maturation is promoted (Kato et al.,
* Correspondence to: Hocheol Kim, Department of Herbal Pharmacology, 2003). Based on these clinical observations, aromatase
of E. senticosus, and the root of Phlomis umbrosa?from College of Oriental Medicine, Kyung Hee University, 1 Hoegidong, inhibition has been proposed as a potential approach
Dongdaemoongu, Seoul 130-701, Republic of Korea. for growth enhancement in children with short stature
Dongeuibogam and designated the formula HT042. The E-mail: hckim@khu.ac.kr (Cernich et al., 2004; Dunkel, 2006).
bone growth effect of HT042 was investigated in adolescent Copyright ??2012 John Wiley & Sons, Ltd. Many ongoing studies are attempting to elucidate al-
ternative therapeutic medicines (Ra et al., 2004). Korean
female Sprague-Dawley rats. We measured the bone growth
Received 30 September 2011
of tibia and BrdU-positive cells on chondrocytes in the Revised 17 January 2012
1376 Accepted 26 January 2012
Molecules 2016, 21, 461; doi:10.3390/molecules21040461 www.mdpi.com/journal/molecules
J. Medicinal food Phytotheraphy research Molecules Molecules
2010 2012 2013 2016
鸚㏝윋?ц뜆耶멩쐝沃???2沅?????2017??1?? ISSN 1229-1765(Print), ISSN 2288-7199(Online) Phytomedicine 32 (2017) 59?67 Hindawi Hindawi
Kor. J. Herbol. 2017竊?2(1)竊?3-68 http://dx.doi.org/10.6116/kjh.2017.32.1.63. Evidence-Based Complementary and Alternative Medicine Evidence-Based Complementary and Alternative Medicine
Contents lists available at ScienceDirect Volume 2017, Article ID 5219418, 7 pages Volume 2017, Article ID 6935802, 8 pages
https://doi.org/10.1155/2017/5219418 https://doi.org/10.1155/2017/6935802
Phytomedicine
Sprague-Dawley怨??곗쪖瑜??댁슜??HT042??14??諛섎났 寃쎄뎄?ъ뿬 ?낆꽦?곌뎄 Research Article Research Article
journal homepage: www.elsevier.com/locate/phymed Astragalus Extract Mixture HT042 Increases Longitudinal Bone
?≪젙鍮?#, ?대룞??#, 源?곸떇1, ?댁듅寃?, 諛곗쭊??, 源?몄쿋1* Astragalus Extract Mixture HT042 Improves Bone Growth Rate by Upregulating Circulatory IGF-1 in Rats
Original Article Growth, Mass, and Microarchitecture in Prepubertal Female
1 : 寃쏀씗??숆탳 ?쒖쓽怨쇰???蹂몄큹?숆탳?? 2 : ?쒖폁??鍮꾩엫?곸뿰援ъ꽱?? Safety evaluation of Astragalus extract mixture HT042 and its Rats: A Microcomputed Tomographic Study Donghun Lee,1 Sung Hyun Lee,2 Yoon Hee Lee,2 Jungbin Song,1 and Hocheol Kim1
constituent herbs in Sprague?Dawley rats 1Department of Herbal Pharmacology, Kyung Hee University College of Korean Medicine, Seoul 02447, Republic of Korea
A 14-day Repeated Dose Oral Toxicity Study of HT042 in Sprague-Dawley Rats Jungbin Song,1 Sung Hyun Lee,2 Donghun Lee,1 and Hocheol Kim1 2Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NeuMed Inc., Seoul 02440, Republic of Korea
Jungbin Song a, Donghun Lee a, Byoungjae Min b, Jin-Sook Bae c, Gyu Tae Chang d, Correspondence should be addressed to Hocheol Kim; hckim@khu.ac.kr
Jungbin Song1#, Donghun Lee1#, Young-Sik Kim1, Seunggyeong Lee1, Hocheol Kim a,? 1Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Received 22 February 2017; Revised 1 May 2017; Accepted 22 May 2017; Published 20 June 2017
Jin-Sook Bae2, Hocheol Kim1* a Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, South Korea Seoul 02447, Republic of Korea Academic Editor: I-Min Liu
b Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Seoul 02440, South Korea 2Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NeuMed Inc., 88 Imun-ro, Dongdaemun-gu, Copyright ??2017 Donghun Lee et al. This is an open access article distributed under the Creative Commons Attribution License,
1 : Dept. of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University c Nonclinical Research Institute, Chemon Inc., 240 Nampyeong-ro, Yangji-myeon, Cheoin-gu, Yongin-si, Gyeonggi-do 17162, South Korea Seoul 02440, Republic of Korea which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
2 : Nonclinical Research Institute, Chemon Inc. d Department of Pediatrics of Korean Medicine, Kyung Hee University Hospital at Gangdong, 892 Dongnam-ro, Gangdong-gu, Seoul 05278, South Korea Correspondence should be addressed to Hocheol Kim; hckim@khu.ac.kr Astragalus extract mixture HT042 is a standardized ingredient of health functional food approved by Korean FDA with a claim
of ?쐆eight growth of children.??HT042 stimulates bone growth rate and increases local IGF-1 expression in growth plate of rats
ABSTRACT article info abstract Received 13 March 2017; Accepted 13 April 2017; Published 10 May 2017 which can be considered as direct stimulation of GH and its paracrine/autocrine actions. However, it remains unclear whether
HT042 stimulates circulatory IGF-1 which also plays a major role to stimulate bone growth. To determine the effects on circulatory
Objectives : HT042 is a combination of three herbal extracts from the roots of Astragalus membranaceus, the stems Article history: Background: Astragalus extract mixture HT042 is a combination of three standardized extracts from As- Academic Editor: Ki-Wan Oh IGF-1, IGF-1 and IGFBP-3 expressions and phosphorylation of JAK2/STAT5 were evaluated in the liver after 10 days of HT042
of Eleutherococcus senticosus and the roots of Phlomis umbrosa, which has been demonstrated to increase longitudinal Received 23 October 2016 tragalus membranaceus root, Eleutherococcus senticosus stem, and Phlomis umbrosa root, which has proven administration. HT042 upregulated liver IGF-1 and IGFBP-3 mRNA expression, IGF-1 protein expression, and phosphorylation
bone growth rate. The aim of this study was to assess the safety of HT042 after repeated oral administration. Revised 20 March 2017 to stimulate children?셲 height growth. Copyright ??2017 Jungbin Song et al. This is an open access article distributed under the Creative Commons Attribution License, of JAK2/STAT5. HT042 also increased bone growth rate and proliferative zonal height in growth plate. In conclusion, HT042
Methods : A 14-day repeated oral dose toxicity study was conducted using male and female Sprague?Dawley rats. Accepted 27 March 2017 Purpose: The aim of this study was to demonstrate the safety of HT042 and its three constituent herbs which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. stimulates bone growth rate via increment of proliferative rate by upregulation of liver IGF-1 and IGFBP-3 mRNA followed by IGF-
HT042 was administered orally at repeated doses of 500, 1,000 and 2,000 ????day for 14 days. Clinical signs and when administered orally. 1 protein expression through phosphorylation of JAK2/STAT5, which can be regarded as normal functioning of GH-dependent
mortality were observed daily, whereas body weight and food consumption were recorded weekly throughout the Keywords: Methods: Acute and sub-chronic oral toxicity studies were conducted using male and female Sprague? Astragalus extract mixture HT042 is a standardized multiherbal mixture comprising Astragalus membranaceus, Eleutherococcus endocrine pathway.
experiment. At the end of the study, blood was taken from the posterior vena cava for hematology and serum Astragalus extract mixture Dawley rats. In the acute toxicity study, HT042 and each of the herbs was administered at single doses of senticosus, and Phlomis umbrosa, which has proven to promote children?셲 height growth. The aim of this study was to investigate
biochemistry. All organs of the body surface, subcutis, head, thoracic cavity, and abdominal cavity were observed HT042 up to 5000 mg/kg. In the 13-week sub-chronic toxicity study, HT042 was administered at repeated doses the effects of HT042 on longitudinal bone growth, bone mass, and bone microstructure in growing rats using a high-resolution
grossly. Then, the internal organs were removed and weighed. Astragalus membranaceus of up to 4000 mg/kg/day. microcomputed tomography system. Four-week-old female rats were fed an HT042-containing diet for 2 weeks. Tibial length was
Results : No death occurred and no significant changes in clinical sign, body weight, food consumption and serum Eleutherococcus senticosus Results: In the acute toxicity study of HT042 and each of the herbs, no deaths occurred and there was measured at baseline and weekly in vivo. At the end of the study, volumetric bone mineral density (vBMD) and microarchitectural
biochemistry parameters were observed in male and female rats over the study period. Although there were some Phlomis umbrosa no indication of toxicity, on the basis of clinical signs, body weight, and necropsy ?ndings. In the sub- parameters were estimated in the trabecular and cortical bone of the tibia. Tibial length gain was significantly increased by HT042
alterations in hematologic and necropsy findings, and organ weights, these changes were not considered toxicologically Toxicity chronic toxicity study of HT042, there were no deaths and no changes in clinical signs or the ?ndings of compared to that reported with the control diet. In the proximal tibial metaphysis, HT042-treated rats had significantly higher
significant. ophthalmic examinations. Although there were some treatment-related changes in other ?ndings, these trabecular vBMD, bone volume fraction, and trabecular number and lower trabecular separation, trabecular pattern factor, and
Conclusions : These results suggest that the 14-day repeated administration of HT042 does not produce any significant alterations were not considered toxicologically signi?cant because they remained within normal ranges structure model index values than control rats did. Total cross-sectional area and bone area of the cortical bone in the tibial diaphysis
oral toxicity at doses of up to 2,000 ????day in male and female rats under the present experimental conditions. or recovered during the recovery period. also increased. These findings suggest that HT042 increases longitudinal bone growth rate, improves trabecular bone mass, and
Conclusion: The oral approximate lethal doses of HT042 and each of the herbs were > 5000 mg/kg, and enhances the microarchitecture of trabecular and cortical bone during growth.
1) the no-observed-adverse-effect level of HT042 was 4000 mg/kg/day in male and female rats.
Key words : HT042, Astragalus membranaceus, Eleutherococcus senticosus, Phlomis umbrosa, toxicity ??2017 Elsevier GmbH. All rights reserved.
?? ??濡? ?랁븳 ?ㅻ뀈??珥덈낯 ?⑷린 Astragalus membranaceus??肉뚮━濡? Introduction The mechanism underlying HT042?셲 activity involves the stimula- 1. Introduction impaired bone structure and bone strength [10]. Growth hor- 1. Introduction [5]. For better understanding of the endocrine versus the
?딂줇?븃〃, ?⒵객易덅뀵, ?섉캃, ?잒굦???⑤뒫?쇰줈 ?ъ슜?섎ʼn, ?먯삤媛 tion of growth hormone secretion (Kim et al., 2013), which is pre- One of the main functions of bone is to provide structural mone (GH) treatment has been reported to increase height, Longitudinal bone growth is the result of chondrocyte pro- paracrine/autocrine roles of IGF-1 in bone growth, previ-
HT042???⑷린, ?먯삤媛 諛??쒖냽??異붿텧臾?蹂듯빀臾쇰줈 怨④만?? (?뷰틪?????먮쫭?섎Т怨?Araliaceae)???랁븳 ?숈뿽愿紐?媛?쒖삤 Astragalus extract mixture HT042 is a combination of three sumed to lead to the systemic production of insulin-like growth support for the body. During childhood, bones grow in size, normalize BMD, and improve bone structure and strength in liferation and ensuing hypertrophy in growth plates, called ous studies have been used to examine the relative con-
?깆옣??珥됱쭊?섎뒗 臾쇱쭏?대떎1-3). ?⑷린(容?)??肄⑷낵(Fabaceae)?? 媛덊뵾 Eleutherococcus senticosus??肉뚮━ 諛?以꾧린濡?獒쒑뀕凉븃뀺, standardized herbal extracts from Astragalus membranaceus root, factor-1 and local production of bone morphogenetic protein-2 and accrue mass, and change their architecture to develop a strong short children [6, 9, 10]. In addition to well-established effects endochondral ossification which is mainly controlled by tribution to longitudinal bone growth. It has been widely
Eleutherococcus senticosus stem, and Phlomis umbrosa root. It has insulin-like growth factor-1 in growth plates (Kim et al., 2010, structure for load bearing [1]. The percentile location of an on growth plate cartilage, GH exerts anabolic effects on bone a system of growth hormone- (GH-) insulin-like growth reported that longitudinal growth is mainly mediated by
*Corresponding author : Hocheol Kim, Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, been developed to promote height growth in children with short 2012). individual?셲 bone traits, such as bone mineral density and and stimulates new bone formation, which results in greater factor-1 (IGF-1) axis [1]. Mammalian growth plate consists of paracrine/autocrine actions of IGF-1 based on the Yakar et al.
26 Kyungheedaero, Dongdaemun-gu, Seoul 02447, Republic of Korea. stature. trabecular and cortical morphology, all of which determine bone mass and improved skeletal structure [11, 12]. three principal zones: resting, proliferative, and hypertrophic study that liver IGF-1 deficient mice developed normally in
쨌Tel竊?82-2-961-0419 쨌E-mail : hckim@khu.ac.kr HT042 is considered safe because each of its constituent herbs bone strength, is likely established during childhood [2?5]. zones. In resting zone, GH begins its actions by direct spite a 75% reduction in serum IGF-1 levels due to compen-
#First author : Jungbin Song, Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 HT042 was found to stimulate height growth in a 3-month, has been widely consumed for a long time and has not shown Therefore, the optimization of bone strength during child- Astragalus extract mixture HT042 is a standardized stimulation of resting stem-like chondrocytes to start pro- satory serum GH and nonhepatic IGF-1 [6, 7]. But, years later,
Kyungheedaero, Dongdaemun-gu, Seoul 02447, Republic of Korea. placebo-controlled trial, and this was further con?rmed in a 6- signi?cant adverse effects (Editorial Board of Zhong Hua Ben hood is important for lifelong bone health. multiherbal mixture consisting of Astragalus membranaceus, liferation. The following cell replication in proliferative zone the same team revealed that circulatory IGF-1 is important
쨌Tel竊?82-2-961-0419 쨌E-mail : jbsong@khu.ac.kr month trial involving 140 children with mild short stature (data Cao, 1999). Nonetheless, even if the herbs are non-toxic when con- Eleutherococcus senticosus, and Phlomis umbrosa. HT042 and terminal differentiation and enlargement in hypertrophic for maintaining normal growth by the study that liver IGF-1
#First author : Donghun Lee, Department of Herbal Pharmacology, College of Korean Medicine, Kyung Hee University, 26 not published). In preclinical studies, HT042 has been shown to sumed individually, their combination could result in toxic effects Several studies have demonstrated that short-statured was found to increase height growth rate in children with zone are both primarily caused by circulatory IGF-1 [2, 3] and and ALS double knockout mice were much smaller because
Kyungheedaero, Dongdaemun-gu, Seoul 02447, Republic of Korea. increase longitudinal bone growth rate through chondrocyte prolif- owing to herb?herb interactions (Che et al., 2013). In addition, al- children have impaired bone health. Low bone mineral mild short stature in a 12-week placebo-controlled trial, locally expressed IGF-1 [4]. there was a decrease in not only IGF-1 production but also
쨌Tel竊?82-2-961-0419 쨌E-mail : allstart2925@naver.com eration and hypertrophy in growth plates (Kim et al., 2010, 2012). though a daily intake of 1500 mg has been proven safe and well- content (BMC) and bone mineral density (BMD) at several which was confirmed by another 24-week trial (data not circulatory IGF-1 stability that originated from compensatory
쨌Received竊?4 November 2016 쨌Revised竊? January 2017 쨌Accepted竊?5 January 2017 tolerated in both 3- and 6-month clinical trials, children may in- skeletal sites are reported in short children [6?9]. Short published). HT042 has been shown to induce longitudinal Circulatory IGF-1 is biosynthesized in the liver as an nonliver sources because of the absence of protective ALS
Abbreviations: RBC, red blood cell; ALT, alanine aminotransferase; ICH, Interna- gest excessive amounts over long periods. Furthermore, unlike the children exhibit increased bone resorption [6] and have bone growth by stimulation of chondrocyte proliferation endocrine hormone in the ternary form bound to IGF protein [3]. This suggests that circulatory IGF-1 plays a major
tional Council for Harmonisation. other two herbs, the systemic toxicity pro?le of P. umbrosa in an- and hypertrophy in growth plates [13, 14]. The mechanism binding protein-3 (IGFBP-3) and acid labile subunit (ALS), role in longitudinal bone growth rate.
imals is not well-established yet. To ensure the safety as an agent while IGF-1 is also expressed by nonliver tissues including
? Corresponding author. growth plate, where it works in a paracrine/autocrine fashion Almost 80% of circulatory IGF-1 is produced by liver in
E-mail address: hckim@khu.ac.kr (H. Kim). humans and rodents. It is well established that circulatory
http://dx.doi.org/10.1016/j.phymed.2017.03.005
0944-7113/??2017 Elsevier GmbH. All rights reserved.
Kor. J. Herbol. Phytomedicine Evidence-Based Complementary Evidence-Based Complementary
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