Abstract
Previous research revealed an increased expression of HSP72 in leukocytes after vigorous endurance exercise. We questioned whether more intensive but shorter exercise also induces leukocyte HSP72 synthesis. To delineate the role of reactive oxygen species (ROS) in exercise-related HSP72 induction, we additionally examined the effect of RRR-α-tocopherol (α-toc) on HSP72 expression using a double-blind placebo (P) controlled cross-over design. After supplementation with α-toc (500 I.U. daily) or P for 8 days, 9 male subjects performed a combined exhaustive treadmill protocol (total duration 29.4 ± 2.0 min). HSP72 was assessed on mRNA (RT-PCR) and protein levels (flow cytometry). HSP72 mRNA rose 3 h after exercise only in the P group, but individual differences (α-toc - P) did not reveal significant treatment effects. A moderate but significant rise of HSP72 protein occurred in granulocytes up to 48 h after exercise. Three hours post-exercise, granulocyte HSP72 protein was lower when subjects received α-toc, but this effect vanished 24 and 48 h post-exercise. Exhaustive treadmill exercise augments HSP72 mRNA in leukocytes and induced a moderate but prolonged response of granulocyte HSP72 protein. These exercise effects are lower when compared to earlier findings obtained after vigorous endurance exercise. ROS seem to be involved, but do not play the major role in the induction of granulocyte HSP72 synthesis after exhaustive exercise.
Key words
HSP72 - exercise - oxidative stress - antioxidants - RT-PCR - flow cytometry - immune function
References
1
Azzi A, Boscobonic D, Hensey C.
The protein kinase family.
Eur J Biochem.
1992;
208
547-557
2
Bachelet M, Mariethoz E, Banzet N, Souil E, Pinot F, Polla C Z, Durand P, Bouchaert P, Polla B S.
Flow cytometry is a rapid and reliable method for evaluating heat shock protein 70 expression in human monocytes.
Cell Stress Chaperones.
1998;
3
168-176
3
Baehner R L, Boxer L A, Allen J M, Davis J.
Autooxidation as a basis for altered function by polymorphonuclear leucocytes.
Blood.
1977;
50
327-335
4
Baehner R L, Boxer L A, Ingraham L M, Butterick C, Haak R A.
The influence of vitamin E on human polymorphonuclear cell metabolism and function.
Ann NY Acad Sci.
1982;
393
237-250
5
Biesalski H K, Greiff H, Brodda K, Hafner G, Baessler K H.
Rapid determination of vitamin A (retinol) and vitamin E (alpha-tocopherol) in human serum by isocratic adsorptions HPLC.
Int J Vit Nutr Res.
1986;
56
319-327
6
Cannon J G, Orencole S F, Fielding R A, Meydani M, Meydani S N, Fiatarone M A, Blumberg J B, Evans W J.
Acute phase response in exercise: interaction of age and vitamin E on neutrophils and muscle enzyme release.
Am J Physiol.
1990;
259
R1214-R1219
7 Dickhuth HH, Huonker M, Münzel T, Drexler H, Berg A, Keul J. Individual anaerobic threshold for evaluation of competitive athletes and patients with left ventricular dysfunction. In: Bachl N, Graham TE, Löllgen H (eds) Advances in Ergometry. Berlin; Springer 1991: 173-179
8
Dill D B, Costill D L.
Calculation of percentage changes in volumes of blood plasma and red cells in dehydration.
J Appl Physiol.
1974;
37
247-248
9
Dimitrov N V, Meyer C, Gilliland D, Ruppenthal M, Chenoweth W, Malone W.
Plasma tocopherol concentrations in response to supplemental vitamin E.
Am J Clin Nutr.
1991;
53
723-729
10
Essig D A, Nosek T M.
Muscle fatigue and induction of stress protein genes: a dual function of reactive oxygen species?.
Can J Appl Physiol.
1997;
22
409-428
11
Fehrenbach E, Niess A M, Schlotz E, Passek F, Dickhuth HH, Northoff H.
Transcriptional and translational regulation of heat shock proteins in leukocytes of endurance runners.
J Appl Physiol.
2000;
89
704-710
12
Fehrenbach E, Niess A M, Veith R, Dickhuth HH, Northoff H.
Changes of HSP72-expression in leukocytes are associated with adaption to exercise under conditions of high environmental temperature.
J Leukoc Biol.
2001;
69
747-754
13
Fincato G, Polentarutti N, Sica A, Mantovani A, Colotta F.
Expression of a heat-inducible gene of the HSP70 family in human myelomonocytic cells: regulation by bacterial products and cytokines.
Blood.
1991;
77
579-586
14 Gardner M J, Altman D G. Statistics with Confidence - Confidence Intervals and Statistical Guidelines. Belfast; Universities Press 1989
15
Gorman A M, Heavey B, Creagh E, Cotter T G, Samali A.
Antioxidant-mediated inhibition of the heat shock response leads to apoptosis.
FEBS Lett.
1999;
445
98-102
16
Guarnieri C, Melandri G, Caldarera I, Cervi V, Semprini F, Branzi A.
Spontaneous superoxide generation by polymorphonuclear leukocytes isolated from patients with stable angina after physical exercise.
Int J Cardiol.
1992;
37
301-307
17 Hartmann A, Niess A M. DNA damage in exercise. In: Sen CK, Packer L, Hanninen O (eds) Handbook of Oxidants and Antioxidants in Exercise. Amsterdam; Elsevier 2000: 195-217
18
Islam K N, Devaraj S, Jialal I.
Alpha-Tocopherol enrichment of monocytes decreases agonist-induced adhesion to human endothelial cells.
Circulation.
1998;
98
2255-2261
19
Jacquier-Sarlin M R, Fuller K, Dinh-Xuan A T, Richard M J, Polla B S.
Protective effects of hsp70 in inflammation.
Experientia.
1994;
50
1031-1038
20
Kelly D A, Tiidus P M, Houston M E, Noble E G.
Effect of vitamin E deprivation and exercise training on induction of HSP70.
J Appl Physiol.
1996;
81
2379-2385
21
Liu Y, Mayr S, Opitz-Gress A, Zeller C, Lormes W, Baur S, Lehmann M, Steinacker J M.
Human skeletal muscle HSP70 response to training in highly trained rowers.
J Appl Physiol.
1999;
86
101-104
22
Locke M.
The cellular stress responses to exercise - Role of stress proteins.
Exerc Sports Sci Rev.
1997;
25
105 - 136
23
Locke M, Noble E G, Atkinson B G.
Exercising mammals synthesize stress proteins.
Am J Physiol.
1990;
258
C723-C729
24
Locke M, Noble E G, Tanguay R M, Feild M R, Ianuzzo S E, Ianuzzo C D.
Activation of heat-shock transcription factor in rat heart after heat shock and exercise.
Am J Physiol.
1995;
268
C1387-C1394
25
Marini M, Frabetti F, Musiani D, Franceschi C.
Oxygen radicals induce stress proteins and tolerance to oxidative stress in human lymphocytes.
Int J Radiat Biol.
1996;
70
337-350
26
McDuffee A T, Senisterra G, Huntley S, Lepock J R, Sekhar K R, Meredith M J, Borrelli M J, Morrow J D, Freeman M L.
Proteins containing non-native disulfide bonds generated by oxidative stress can act as signals for the induction of the heat shock response.
J Cell Physiol.
1997;
171
143-151
27
Meydani M.
Vitamin E.
Lancet.
1995;
345
170-175
28
Mizzen L A, Welch W J.
Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heat-shock protein 70 expression.
J Cell Biol.
1988;
106
1105-1116
29
Moseley P L.
Exercise, stress, and the immune conversation.
Exerc Sports Sci Rev.
2000;
28
128-132
30
Niess A M, Dickhuth HH, Northoff H, Fehrenbach E.
Free radicals and oxidative stress in exercise - immunological aspects.
Exerc Immunol Rev.
1999;
5
22-56
31
Niess A M, Passek F, Lorenz I, Schneider E M, Dickhuth HH, Northoff H, Fehrenbach E.
Expression of the antioxidant stress protein heme oxygenase-1 (HO-1) in human leukocytes - Acute and adaptional responses to endurance exercise.
Free Rad Biol Med.
1999;
26
184-192
32
Noble E G, Moraska A, Mazzeo R S, Roth D A, Olsson M C, Moore R L, Fleshner M.
Differential expression of stress proteins in rat myocardium after wheel or treadmill run training.
J Appl Physiol.
1999;
86
1696-1701
33
Paroo Z, Tiidus P M, Noble E G.
Estrogen attenuates HSP72 expression in acutely exercised male rodents.
Eur J Appl Physiol.
1999;
80
180-184
34
Pinot F, el-Yaagoubi A, Christie P, Dinh-Xuan A T, Polla B S.
Induction of stress proteins by tobacco smoke in human monocytes: modulation by antioxidants.
Cell Stress Chaperones.
1997;
2
156-161
35
Polla B S, Perin M, Pizurki L.
Regulation and functions of stress proteins in allergy and inflammation.
Clin Exp Allergy.
1993;
23
548-556
36
Polla B S, Stubbe H, Kantengwa S, Maridonneau-Parini I, Jacquier-Sarlin M R.
Differential induction of stress proteins and functional effects of heat shock in human phagocytes.
Inflammation.
1995;
19
363-378
37
Roecker K, Schotte O, Niess A M, Horstmann T, Dickhuth H H.
Predicting competition performance in long-distance running by means of a treadmill test.
Med Sci Sports Exerc.
1998;
30
1552-1557
38
Salo D C, Donovan C M, Davies K J.
HSP70 and other possible heat shock or oxidative stress proteins are induced in skeletal muscle, heart, and liver during exercise.
Free Radic Biol Med.
1991;
11
239-246
39 Traber M G. Vitamin E. In: Sen CK, Packer L, Hanninen O (eds) Handbook of Oxidants and Antioxidants in Exercise. Amsterdam; Elsevier 2000: 359-371
40
Trautinger F, Kokesch C, Klosner G, Knobler R M, Kindas-Mugge I.
Expression of the 72-kD heat shock protein is induced by ultraviolet A radiation in a human fibrosarcoma cell line.
Exp Dermatol.
1999;
8
187-192
41
Ushakova T, Melkonyan H, Nikonova L, Mudrik N, Gogvadze V, Zhukova A, Gaziev A I, Bradbury R.
The effect of dietary supplements on gene expression in mice tissues.
Free Radic Biol Med.
1996;
20
279-284
42
Vigh L, Maresca B, Harwood J L.
Does the membrane's physical state control the expression of heat shock and other genes?.
Trends Biochem Sci.
1998;
23
369-374
43
Wei Q, Xu X, Cheng L, Legerski R J, Ali-Osman F.
Simultaneous amplification of four DNA repair genes and ß-actin in human lymphocytes by multiplex reverse transcriptase-PCR.
Cancer Res.
1995;
55
5025-5029
44
Zhao Y, Wein A J, Levin R M.
Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method.
Mol Cell Biochem.
1995;
148
1-7
A. M. Niess, M.D.
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