modulation of neuroendocrinological function by psychosomatic training: acute effect of chundosunbup...

Psychoneuroendocrinology 25 (2000) 439–451www.elsevier.com/locate/psyneuen

Modulation of neuroendocrinological functionby psychosomatic training: acute effect of

ChunDoSunBupQi-training on growthhormone, insulin-like growth factor (IGF)-I, and

insulin-like growth factor binding protein(IGFBP)-3 in men

Hoon Ryua,b,*, Myeong-Soo Leea, Seong-Min Jeonga,Jung-Hee Leeb, Chang-Won Kangc, Dae-Yeol Leed,

Hun-Taeg Chungb

a Department of Immunology and Qi (Bioenergy) Medicine, Institute of Biotechnology, WonkwangUniversity, Iksan, Chonbuk, South Korea

b Department of Microbiology and Immunology, School of Medicine, Wonkwang University, Iksan,Chonbuk 570-749, South Korea

c Department of Physiology, College of Veterinary Medicine, Chonbuk National University, Chonbuk,South Korea

d Department of Pediatrics, Medical School, Chonbuk National University, Iksan, Chonbuk, SouthKorea

Received 2 July 1999; accepted 5 November 1999

Abstract

The neuroendocrine system is regarded as the major link between the psychological stateand the immune system in man. The present study was undertaken to examine the acute effectof ChunDoSunBupQi-training, a traditional psychosomatic training, on the plasma level ofgrowth hormone (GH), insulin-like growth factor (IGF)-I and insulin like growth factor binding

* Corresponding author. Present address: Department of Neurology, Harvard Institutes of Medicine,Room 847, 77 Avenue Louis Pasteur, Boston, MA 02115, USA. Tel.:+1-617-6670832; fax:+1-617-6670800.

E-mail address:[email protected] (H. Ryu) or [email protected] (H.-T. Chang).

0306-4530/00/$ - see front matter 2000 Elsevier Science Ltd. All rights reserved.PII: S0306 -4530(99 )00069-4

440 H. Ryu et al. / Psychoneuroendocrinology 25 (2000) 439–451

protein (IGFBP)-3 in young men. To characterize the plasma level of hormones and the patternof IGFBP-3 at pre- (210 min), mid-time (40 min), and immediately after (post-time,+10 min)Qi-training, radioimmunoassay (RIA) and western ligand blot (WLB) analysis were used. Theplasma level of GH at the mid-time was significantly increased (P,0.05) compared to thelevel at pre-time of Qi-training. The plasma level of IGF-I was also increased at mid-time(P,0.05) and immediately after Qi-training. There was a significant correlation between thelevels of GH and IGF-I immediately after Qi-training (r=0.69, P,0.01). In this study, thechange of IGFBP-3 among the several IGFBPs was determined. There was a relative shift inIGFBP-3 for the 43-kDa fraction during mid-time of Qi-training (P,0.05). These results sug-gest that Qi-training influences the growth hormone, IGF-I and IGFBP-3 status via brain-targetorgan axis in men. 2000 Elsevier Science Ltd. All rights reserved.

Keywords:Growth hormone; Insulin-like growth factor (IGF)-I; IGF-binding protein (IGFBP)-3;Neuroendocrine-immune axis; Qi-training

1. Introduction

It is well known that the psychological condition can modulates immunologicalfunctioning via neurohormones (Besedovsky and Sorkin, 1977). On the basis of thepsycho-neuroendocrine-immune circuits, the in vivo effect of psychosomatic inter-vention has been investigated (Liu et al., 1990; Jin, 1992; Weidemann et al., 1994;Ryu et al., 1995a,b; Lee et al., 1997). Among various kinds of interventions, Qi-training and its beneficial effects has received increasing attention due to the linkbetween psychological stabilization and immune function. Qi-training is an orientaltraditional training that not only develops physical balance through isotonic and iso-metric slow motions but also perpetuates psychological stabilization through medi-tation (Jin, 1992; Ryu et al., 1996a; Lee et al., 1997, 1998). The uniqueness of Qi-training has been distinguished from physical exercise (Liu et al., 1990; Jin, 1992;Ryu et al., 1995a; Kim et al., 1996). Several studies have found that neuronal activityand cardiovascular function are modulated by Qi-training (Liu et al., 1990; Lim etal., 1993). Recently we have investigated the acute and chronic effects of Qi-training(Ryu et al., 1995a,b, 1996a; Lee et al., 1997). It is possible that at least part ofthe beneficial effects of Qi-training on psychological and immunological functionsincluding cardiovascular function may be mediated via neuroendocrine responses(Ryu et al., 1995a,b, 1996a,b). Although there are a number of positive findings inpsycho-physiological and immunological functions, there have been few systematicinvestigations of the precise neuroendocrine mechanisms by which Qi-training maymoderate immunological functions.

Endogenous neuropeptides are capable of interacting with immune organs andcells. Among anterior pituitary hormones, growth hormone (GH) is known toundergo large changes in circulating concentration in response to several stimuli suchas physical exercise, sleep and fasting (Weidemann et al., 1994). Growth hormonestimulates the liver and other organs, including the skeleton, to synthesize and secreteinsulin like growth factors (IGFs) (Yeh et al., 1994).

IGF-I and IGF-II are the circulating polypeptides traditionally thought to mediate

441H. Ryu et al. / Psychoneuroendocrinology 25 (2000) 439–451

GH-induced longitudinal growth (Harel and Tannenbaum, 1992). Plasma IGF-I lev-els have been correlated with the integrated 24-h secretion of GH. Since plasmaIGF-I levels lack circadian fluctuations, they are considered better measures of theendogenous average secretion of GH (Kssem et al., 1994). IGF-I has been demon-strated to stimulate osteoblast proliferation and collagen synthesis in vitro. A familyof IGF-binding proteins (IGFBPs) have been shown to modulate the actions of IGF-I. IGF-I and IGFBP-3 are used as the virtually invariable indices of GH secretionin clinical practice (Hasegawa et al., 1994).

Studies have shown that physical exercise changes somatic growth and develop-ment by triggering the pulsatile secretion of growth hormones (De La Fuente andWells, 1981; Harel and Tannenbaum, 1992; Yeh et al., 1994). Mechanical loading,muscular activity, and gravity serve as extracellular stimuli that are transmitted tothe pituitary and other organs, and participate in the endocrine regulation for growth(Weidemann et al., 1994; Yeh et al., 1994). Moderate exercise, rather than intensivephysical exercise, has several positive effects such as maintaining active bone forma-tion and reducing bone resorption through the release of growth factors in variousanimal models (Yeh and Aloia, 1990; Yeh et al., 1994). Thus, many investigatorshave reported the influence of physical exercise on growth factors; however, themodulation of growth factors by Qi-training has not been determined (Poehlman etal., 1990, 1994; Weidemann et al., 1994; Yeh et al., 1994).

Therefore, the present study was designed to examine the acute effect of CDSBQi-training on the plasma levels of GH and IGF-I, and the pattern of IGFBP-3 inyoung males. We also intended to investigate the possibly expected correlation ofthe circulating level of GH and IGF-I and determine whether the association ofpituitary and target organ axis would be apparent in GH-induced IGF-I secretionin men.

2. Materials and methods

2.1. Subjects in hormonal experiments

The investigations were carried out on a group engaged inChunDoSunBup(CDSB) Qi-training in the CDSB Center and several branches in Seoul, South Korea.A total of 16 healthy young men (mean age, 29) agreed to participate in the studyand signed an informed consent form obtained from the Human Subjects ReviewBoard in Wonkwang University Hospital and School of Medicine. All trainees hadpracticed Qi-training for 6 months. Sedentary control subjects (n=10, mean age, 26)were recruited by advertisement through the Wonkwang University for participationin the study. All subjects who participated in this study were in good health withouta history of chronic disease, malnutrition, malignancy or renal disease. They had notreceived any medications such as steroid hormone that might affect physical activity.


2.2. Self-report measures

The Beck Depression Inventory (BDI) was administered each time blood wasdrawn from the experimental subjects. The BDI is a 21-item questionnaire con-structed to assess the depth and severity of depression. Subjects that scored 17 orhigher on the BDI were assessed as clinically depressed. Since research has shownthat depression suppresses the endocrine system, subjects who scored in the clinicallydepressed range were not included in the analysis. Also, subjects who experiencedhighly stressful events during the study were not included in the results (five subjectswere rejected).

2.3. Experimental procedure and blood sampling

ChunDoSunBup(CDSB)Qi-training which has previously been reported by Ryuet al. (1995a) was performed by the subjects for 1 h and was directed by a master.For 1 h of Qi-training, resting for 10 min before Qi-training and three kinds ofexercise [sound (Chunmoon) reciting for 15 min, slow motions for 15 min, andmeditation for 20 min] were performed. At first participants began the Qi-trainingin a sitting position by reciting vibrational sounds which have no meaning. Thesounds consisted of eight lines as follows: (1) Soo Soo Ma Di Di Di Si; (2) Eh DeDe De Si; (3) Kke De Ma Di Wa; (4) Se Ne Don Di Mi; (5) Kye De Kye Si Mi;(6) Kka Da Wa Ri Ba Si Ma; (7) Wa Na Wat Da Wa Si; and (8) Oh Me Geu BiGeu Bi Sa Ba Si Bi Si. Subjects recited each line very clearly and evenly in onebreath for 15 min. After reciting the sounds, the subjects performed the trainingmotions in a standing position for 15 min. The subjects placed both feet parallel,shoulder width apart, both arms were stretched from the abdomen (Danjun) up toshoulder level. Focusing on the finger tips and center of the palms, the subjectspulled Qi from the Symbol of Truth for 5 min. Next, starting from the abdomen,the subjects raised both arms from the sides of the body until horizontal to shoulderlevel. The hands were then turned slowly until the palms faced upwards to gatherYang (Chun)-Qi until both arms touched the ears. Yang-Qi was pull down throughthe crown of the head, face, chest, and into the abdomen. Again starting from theabdomen both arms were raised from the sides until horizontal to shoulder level.The trunk was bent slowly to gather Yin (Ji)-Qi with both hands; the trunk wasraised while pulling up Yin-Qi through the feet, legs, and into the abdomen. Finally,the subjects sat on the floor with back straight, hands facing upward on the knees,the eyes closed and meditated for 20 min.

The peripheral blood was obtained by venipuncture using heparinized (10 U/ml)syringes from the median cubital vein. In order to determine the optimal bleedingtimes, three trained individuals participated in CDSB Qi-training. From preliminarydata, three bleeding time points were selected for all subsequent experiments to min-imize the number of blood drawings (Ryu et al., 1996a,b). Blood was drawn at rest,pre-time of Qi-training [that is 10 min before Qi-training (210 min)], at 40 min (themid-time of Qi-training, the time between motions and meditation), and immediatelyafter the Qi-training [within 10 min (+10 min)]. Peripheral blood was placed into


pre-chilled tubes containing 200 Kallikrein inhibitor units/ml aprotinin, 1 mM phe-nylmethylsulfonyl-fluoride (PMSF), 10 mM ethylenediamine tetraacetic acid(EDTA), and 50 benzoyl arginine ethyl ester unit/ml soybean trypsin inhibitor(SBTI). The samples were immediately centrifuged at 3000 rpm for 15 min at 4°C,the plasma was separated and stored at220°C until assayed.

2.4. RIA of GH

The plasma level of GH was measured by liquid phase double-antibody125I RIAkit using materials supplied by the DPC (Diagnostic Products Corporation, Los Ang-eles, CA, USA). The intra- and interassay coefficients of variation were 5 and11%, respectively.

2.5. Iodination of IGF-I

The plasma level of IGF-I was measured by radioimmunoassay as previouslydescribed (Lee et al., 1993). IGF-I was iodinated by a modification of the chloramineT method. A 1µg of IGF-I was dissolved in 10µl of 0.2 M sodium phosphatebuffer in a microcentrifuge tube and 10µl (1 mCi) of 125I were added to the sametube containing IGF-I. After the treatment of 10µl (40 µg) of chloramine-T, thetube was incubated for 30 s at room temperature. To stop the reaction, 200µl ofbovine serum albumin (BSA) (30%) instead of sodium metabisulfite were added.The labeled IGF-I was purified by applying the reaction mixture to a column ofSephadex G-50. The labeled compound was eluted with 0.1 N acetic acid containing0.3% BSA, 0.1% glycine and 1000 kallikrein inhibitor units (KIU)/ml aprotinin.Specific radioactivity of the purified radioiodinated IGF-I was 400–700µCi/µg. Thepurified labeled IGF-I was aliquoted as 100 000 cpm/tube and stored at deep freezer.

2.6. Radioimmunoassay of IGF-I

Before the RIA of IGF-I plasma, samples were extracted by using Sep-pak C18cartridges (Waters, MA, USA) to separate IGF-I from IGFBPs. In brief, 100µl ofplasma were preincubated with 650µl of 1% trifluoroacetic acid (TFA, Sigma, MO,USA) for 20 min. Samples were then applied on a Sep-Pak C18 cartridge previouslyactivated with 3 ml of 100% acetonitrile and 0.1% TFA. The cartridge was rewashedwith 3 ml of 0.1% TFA and the absorbed peptide was eluted with 3 ml of 60%acetonitrile in 0.5% TFA. The eluent was subsequently dried using a Speed-Vacconcentrator (Savant, Hicksville, NY, USA). The recovery rate of125I-IGF-I by aSep-Pak C18 cartridge was 86%. The radioimmunoassay was performed in Tris-acetate buffer (0.1 M, pH 7.4) containing 0.2% neomycin, 10 mM EDTA, 50 benzoylarginine ethyl ester units/ml soybean trypsin inhibitor, 200 KIU/ml aprotinin, 1 mMphenylmethylsulfonyl-fluororide, 0.02% sodium azide and 1% BSA. The volume ofthe sample used for the radioimmunoassay was 50µl and the total assay volumewas 300µl. The bound form was separated from free form by Dextran-charcoalsuspension [(0.6% charcoal, 0.06% Dextran T-70, 34µg/ml phenylmercuric acetate,


0.2% neomycin in 0.1 M Tris-acetate buffer (pH 7.4)]. Standard or samples wereincubated with 100µl of diluted antiserum for 24 h at 4°C. Following the additionalincubation with125I-IGF-I for 24 h at 4°C, separation of free form of tracer fromantibody-bound125I-IGF-I complex was achieved by adding 1 ml of Dextran-charcoalsuspension. The least detectable quantity of IGF-I was 10 pg/tube. The intraassayvariation for plasma IGF-I was less than 10% and the interassay variation was lessthan 9%.

2.7. Western ligand blot (WLB) analysis

Three microliters of serum were electrophoresed on 10% SDS-PAGE under a non-reducing condition. The proteins were electrophoretically transferred to nitrocellulosemembranes by applying a constant voltage of 20 V for approximately 16 h. Thetransfer buffer consisted of 25 mM Tris, 190 mM glycine, and 20% methanol. Aspreviously described by Hossenlopp et al. (1990), the membranes were dried for 5min at 37°C and consecutively washed in a Western ligand blot (WLB) buffer con-sisting of 150 mM NaCl, 10 mM Tris–HCl (pH 7.4), 0.5 mg/ml sodium azide sup-plemented first with 3% Nonidet P-40, second with 1% BSA, and third with 0.1%Tween-20. The membranes were incubated overnight with 2×106 cpm of [125I]-IGF-I, 1% BSA, and 0.1% Tween-20 and washed in WLB buffer containing 0.1% tween-20 and then in WLB buffer alone. The membranes were blotted dry and exposed toX-ray films for 7 days according to the method of Hossenlopp et al. (1990). TheIGFBPs were visualized by autoradiography. The autoradiographs of IGFBP-3 ligandblots were scanned using a laser densitometer (PC model, Szimato, Tokyo, Japan).The relative densities were expressed as arbitrary absorbance units per mm.

2.8. Reagent

Recombinant human IGF-I was purchased from Bachem (Torance, CA, USA).Polyclonal anti-somatomedine-C antiserum, a generous gift of Drs L.E. Underwoodand J.J. Van Wyk (University of North Carolina at Chapel Hill), was distributedthrough the hormone Distribution Program of the NIDDK of the National Hormoneand Pituitary Program. Aprotinin, bovine serum albumin (BSA), charcoal, chloram-ine-T, EDTA, glycine, lysozyme, neomycin, phenylmercuric acetate, phenylmethyl-sulfonylfluoride, sodium azide, soybean trypsin inhibitor, Trizma, pyruvate and otherchemicals were purchased from Sigma Chemical (St. Louis, MO, USA). Dextran-T70 and Sephadex G-50 were purchased from Pharmacia Fine Chemicals and iodine-125 was purchased from Amersham International (Buckinghamshire, UK).

2.9. Statistical analysis

Data is presented as the average±SEM. ANOVA and post-hoc test were used tocompare differences at the pre-, mid- and post-time of Qi-training in all demographicand descriptive data. Correlations between hormones were evaluated by regressionanalysis (two-tailed). Results were considered significant at the 95% confidence lim-


its when P,0.05. A computer program (GB-STAT, Dynamic Mycrosystem, Inc.Silver Spring, MD) was use for all statistical calculations.

3. Results

3.1. Effect of CDSB Qi-training on the plasma level of growth hormone (GH)

Fig. 1A shows that the plasma GH level was increased at the mid-time (40 min)(2.16+0.48 ng/ml) (F[2,30]=4.89 P,0.05, by ANOVA and post hoc test) comparedto the pre-time of Qi-training (1.26±0.37 ng/ml). The plasma GH level at the post-time (+10 min) of the Qi-training (2.18±0.56 ng/ml) was also maintained higher asmuch as values of the mid-time, but there was no significance compared to pre-time(210 min) of Qi-training. In separate experiments, it was observed that the level ofGH returned to the basal level 2 h after CDSB Qi-training, the same as before Qi-training (data not shown). To compare the effect of Qi-training on GH level, wemeasured the GH level from the sedentary (resting) group (n=10). The basal levelof GH of the resting group was 1.24±0.34 ng/ml. There was no difference in the basallevel of GH between the Qi-training group and the sedentary group. Furthermore, the40 min resting (1.04±0.25 ng/ml) and 70 min resting (1.12±0.13 ng/ml) did not showany significant changes in the GH level.

3.2. Effect of CDSB Qi-training on the plasma level of insulin-like growth factor(IGF)-I

The plasma level of IGF-I was significantly increased at the mid-time(286.40±15.97 ng/ml) (F[2,30]=3.75P,0.05) compared to the pre-time of Qi-train-

Fig. 1. Effect of Qi-training on plasma levels of growth hormone (GH) (A) and insulin like growthfactor (IGF)-I (B). Pre-time is 10 min before Qi-training (210 min), mid-time is 40 min during Qi-training, and post-time is immediately after Qi-training within 10 min (+10 min). Values are mean±SE(n=16). * Significantly different from values at pre-time of Qi-training by ANOVA and post hoc test,P,0.05.


ing (245.57±13.90 ng/ml) (Fig. 1B). The plasma level of IGF-I at the post-time ofQi-training 287.02±23.13 ng/ml) was higher than that at the mid-time of training butthere was no significant difference (P,0.09). In separate experiments, it was alsoobserved that the level of IGF-I recovered to the basal level 2 h after CDSB Qi-training, the same as before Qi-training (data not shown).

3.3. Effect of CDSB Qi-training on the plasma level of insulin-like growth factorbinding protein (IGFBP)-3

Western ligand blot (WLB) analysis was performed in order to measure the plasmalevel of IGFBPs. In a test for the characterization of patterns of IGFBPs in plasma,the relative shift in the binding intensities for the IGFBP-3 for the 43-kDa fractionduring mid-time [10 min (lane 3), 40 min (lane 4), and 60 min (lane 5)] of Qi-training was found (Fig. 2). Among the IGFBPs there was a focus on the relativeintensity (RI) of [125I]-IGF-I for IGFBP-3. Not only was the IGFBP-3 the most abun-dant BP in plasma but the level of IGFBP-3 as well as IGF-I was also dependenton GH secretion. In 16 samplings, the circulating levels of IGFBP-3 in plasma weremarkedly increased in mid-time (40 min) compares to pre-time (210 min) of Qi-training (Fig. 3). The change of autoradiographic density was confirmed by densitro-metric analysis (Table 1). The relative densitometric value of IGFBP-3 was statisti-cally significant in mid-time (40 min) than in pre-time (210 min) of Qi-training(F[2, 30]=3.83,P,0.05) (Table 1). But the relative densitometric value in IGFBP-3 at post-time of QI-training returned to the basal level in subjects.

In order to examine the modulation of GH-IGF-I axis by CDSB Qi-training, thelinear correlation between the plasma level of GH and IGF-I was evaluated to deter-mine the functional linkage of GH-IGF-I axis. No significant correlation existed atboth the pre- (r=0.01,P,0.99) and mid-time of Qi-training (r=0.18,P,0.50). How-ever, there was the positive correlation at post-time of Qi-training which was statisti-cally significant (r=0.69, P,0.01). In our experiment, otherwise, either the corre-lation between the level of GH and IGFBP-3 (r=0.09, P,0.73), and the level of

Fig. 2. Western ligand blot analysis of IGF-binding proteins (IGFBPs) pattern in plasma of a normalhealthy subject (lane 1) and a Qi-training subject (lanes 2–6). Lane 2, 10 min before (210 min) Qi-training; lane 3, 10 min mid-time of Qi-training; lane 4, 40 min mid-time of Qi-training; lane 5, immedi-ately after Qi-training (within 10 min); lane 6, 60 min after Qi-training. Altered IGFBPs pattern duringQi-training is shown in lane 2–5. The IGFBPs appear from above in the following order: IGFBP-3;IGFBP-2; IGFBP-1; and IGFBP-4. Molecular weight is in kilodaltons (kDa).


Fig. 3. Effect of Qi-training on the plasma IGFBP-3 pattern. Changes of the typical IGFBP-3 patternfrom 16 subjects are represented. Each three lanes represent pre- (a) , mid- (b) and post-time (c) of Qi-training, respectively. Molecular weight in 43-kilodaltons (kDa) indicates IGFBP-3.

Table 1Changes of the level of IGFBP-3 by Qi-traininga

Pre-time (210 min) Mid-time (40 min) Post-time (+10 min)

Arbitrary (unit/mm) 128.19±10.15 140.69±9.14b 125.94±10.20

a Values are mean±SE (n=16) The data is derived from densitometric analysis of autoradiographicdata from Fig. 3.

b Significantly different from values at pre-time of Qi-training by ANOVA and post hoc test,P,0.05.

IGF-I and IGFBP-3 (r=0.21,P,0.44) at the mid-time of Qi-training (40 min) werenot significant.

4. Discussion

The current findings of the increased plasma level of GH, IGF-I and IGF bindingprotein during and after Qi-training indicate that growth factors are acutely triggeredby a traditional meditative exercise training. GH is a classical anterior pituitary hor-mone that regulates the promotion of cell growth and metabolism (Kappel et al.,1993; Thelen et al., 1993). Furthermore, GH has been shown to influence the devel-opment and function of the immune system (Spadoni et al., 1991; Wiedermann etal., 1991a). A novel function of GH was also found for macrophages in vivo andin neutrophils in vitro employing native purified and recombinant GH (Wiedermannet al., 1991b; Ryu et al., 1997). Weidemann et al. (1994) observed that GH wasreleased at a very low threshold of exercise intensity as it increased four to fivefoldafter 8 min of very light ‘warm-up’ exercise. It is clear from many studies (Jahreiset al., 1989; Weidemann et al., 1994; Yeh et al., 1994) that GH secretion respondscontinuously to physical exercise over a wide range of intensities. The possible role


of GH secreted by the stimulation of physical exercise has been investigated inimmune cells (Weidemann et al., 1994).

Qi-training is distinguished from physical exercise in its unique composition, forinstance, the exercise intensity, the word chanting, and the meditation (Ryu et al.,1995a,b, 1996a; Lee et al., 1997). Kim et al. (1996) has reported that Qi-trainingexhibits a low exercise intensity with 13% VO2max in young men. Our previousstudy shows that Qi-training is sufficient to elevate the capacity of circulating neutro-phils to produce microbicidal reactive oxygen intermediate (superoxide anion (O2-)) upon stimulation in aged men (Ryu et al., 1996b). This respiratory function isquickly and markedly activated in the isolated neutrophil at the mid-time andimmediately after Qi-training when compared to values before Qi-training. Takentogether, the previous and present data support that endogenous GH, which issecreted by Qi-training as well as moderate exercise, may enhance resistance toinfection and inflammation through the modulation of innate defense function (Smithet al., 1990; Weidemann et al., 1994; Ryu et al., 1995b).

It appears likely that moderate exercise serves not only as a systemic stimuli butalso as a localized stimuli that is transmitted to localized tissues or cells to initiatetheir genetic program for growth and differentiation. Furthermore, it is suggestedthat the cellular events by mechanical stress are mediated through autocrine or parac-rine factors that are generated in the microenvironment of the tissue (Frost, 1992).It is possible that IGFs are one of these factors in bone formation (Kelly et al., 1990;Yeh et al., 1994). IGF-I is a single chain 7.5-kDa polypeptide that mediates theanabolic effect of GH. Physical exercise increases the plasma and tibial concentrationof IGF-I in rats (Yeh et al., 1994). It is well known that the primary source of plasmaIGF-I is the liver (Harel and Tannenbaum, 1992) and GH administration stimulatesliver production of IGF-I. However, the source of increased plasma levels of IGF-I during and after exercise is not known. Certainly, exercise may stimulate GHsecretion from the anterior pituitary (Weidemann et al., 1994), thereby raising liverproduction of IGF-I, but the possibility of other organs including the skeleton, releas-ing IGF-I cannot be excluded. The immunoregulatory properties of IGF-I as well asGH have been studied (Bjerknes et al., 1997). Bjerknes et al. (1997) demonstratedthat IGF-I is a potent regulator of mature neutrophil function. In the present study,the positive correlation of plasma levels of GH and IGF-I immediately after the Qi-training strongly suggested that these growth factors might function independentlyor synergistically with in vivo. This possibility requires further investigation.

IGFBP-3 is a 42-kDa to 45-kDa molecular weight protein that binds most of IGFsand circulates in plasma as a 150-kDa complex consisting of IGF-I or II, IGFBP-3,and acid labile sub-unit (Jahreis et al., 1991; Rosen et al., 1992; Kssem et al., 1994;Lee et al., 1994a,b). IGFBP-3 is the most abundant BP in the human plasma, butits primary production site remains uncertain. The plasma level of IGFBP-3 is depen-dent on GH secretion (Baxter and Cowell, 1987; Donahue et al., 1990). IGFs boundin IGFBP complexes are unable to bind their target cell membrane receptors (Leeet al., 1993, 1994a). For those situations in which IGFs bound to IGFBPs are biologi-cally inactive, dissociation of IGF and BP complex would be necessary for IGFbioactivity to occur (Lee et al., 1993). In this study, we determined the plasma level


of IGFBP-3 in subjects practicing Qi-training by western-ligand blot (WLB) analysis.The autoradiographic relative intensity of plasma IGFBP-3 was increased signifi-cantly at the mid time of Qi-training. In agreement with previous studies, this resultindicates that the circulating level of IGFBP-3 as well as IGF-I reflects the pulsatileGH peak in young males. Additional research is now required to determine whichaspects of Qi-training contributed to these changes in growth factors, and to acertainwhether exercise in general would result in similar alterations, or if they were aug-mented by the traditional meditative aspects of this Korean exercise.

In conclusion, our present study shows how regular Qi-training acutely effects thesecretion of growth factors in young males. Both GH and IGF-I increased at themid-time and the post-time of Qi-training. The increase in the plasma level of IGF-I was associated with the secretion of GH. The plasma level of IGFBP-3 was alsomodulated through Qi-training. These results suggest that Qi-training is a potentialmethod modulating the secretion of growth factors in men.

Acknowledgements

We thank Won-Bak Kim for his fine technical assistance. Luis Castellano andElizabeth A. Hayes and Nancy K. Hurley are greatly acknowledged for their assist-ance in the preparation of this manuscript. We thank the Masters and trainees inChunDoSunBupQi-training Center who anticipated in the present study. This studywas supported by Post-Doctoral Fellowship (H.R.) from Korea Science and Engin-eering Foundation and a grant (H.T.C.) from Korea Ministry of Education.

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