Effects of Flaxseed Lignans on the Growth of Skeletal Muscle of Male Rats and Its Possible Mechanism

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1 2009,42(1): Scientia Agricultura Sinica doi: /j.issn Effects of Flaxseed Lignans on the Growth of Skeletal Muscle of Male Rats and Its Possible Mechanism ZHOU Wei, WANG Guo-jie, HAN Zheng-kang (Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture of China, Nanjing Agricultural University, Nanjing ) Abstract: Objective To determine whether flaxseed lignans could affect the growth of skeletal muscle in male animals and its possible mechanisms. Method The effect of the flaxseed lignans on skeletal muscle in male rats were determined in vivo. Daidzein (5 mg kg -1 ) and flaxseed lignans (50 mg kg -1 ) were added into the basal diets, respectively, and the concentrations of the serum lignans and daidzein were measured by HPLC, serum growth hormone and testosterone (T) levels were analyzed by RIA, and the expression of estrogen receptor β (ERβ) in the soleus muscle and hypothalamus were determined by RT-PCR. Result Flaxseed lignans and daidzein significantly improved the feed efficiency and increased the total weight gain of the femoral muscle in male rats. The rate of RNA to DNA in muscles and serum T levels were remarkably increased, while the urea nitrogen concentrations were significantly decreased by daidzein and flaxseed lignan and/or its metabolites. Meanwhile the expression of estrogen receptorβ (ERβ) in soleus muscle and hypothalamus were both upgraded by the two phytoestrogens. Conclusion Flaxseed lignan promoted the growth of male rats, and it may regulate serum T levels through binding to ER β in hypothalamus. In turn, it depressed the catabolism of protein and promoted the hypertrophy of skeletal muscle cells. Key words: phytoestrogens; flaxseed lignans; daidzein; mammalian lignans; skeletal muscle growth isoflavoneslignans 2 13 mg g t Zhouwei0732@sohu.com 1924 zhengkanghan@163.com

2 [1] secoisolariciresinol diglusoside SDG Mammalian lignans [2] enterolactone, ENL enterodiol END [3] [4] [5] [6] [7] [8] daidzein Da Sprague-Dawley SD Andersson [9] 1 1 V V 48 h 1.6% 2 mol L -1 ph mm Kieselgel V V V HPLC Agilent 1100 Series 95.2% 28 d SD Sprague-Dawley g SLAC 36 SD mg kg -1 SDG 5 mg kg % 35% 20% 5% 1% 1% 1% 1% 1% 1.0 g STS [10] β estrogen receptorβ ERβ mrna RT-PCR ERβcDNA AF Primer Primer 5.0 [11] 5 -TGT CCA GCC ACG AAT CAG-3 5 -CCT ACC TCC ACG ATT ACC-3 GAPDH 3- [12] 5 -CTT CCG TGC GAG CAC TGC-3 5 -CAT TAT GGC GTG TGC AG-3 PCR 2.5 µl 10 PCR Buffer 1.5 µl 1.5 mmol L -1 MgCl 2 2 µl 0.2 mmol L -1 dntp 2 µl RT 1 µl 0.5 µl 13.8 µl ddh 2 O 5U Taq DNA 95 5 min s s s min SDG 1 ml 1 ml r/min 4 30 min 200 µl SDG 1 ml 37 [7] [13] Spherisorb ODS2 4.6 mmi.d. 150 mm 5 µm Waters, 1 ml min -1 0 min 20 min 10% 90% 280 nm 8 nm µl Bowey [14] SPSS t Mean±SD

3 SDG SDG SDG ENL END SDG 2 SDG 12.44% P % P % P % P mg ml -1 Table 1 The concentrations of serum lignans and daidzein (mg ml -1 ) Control SDG treatment Da treatment SDG UD 0.072±0.018 UD END UD 0.019±0.006 UD ENL UD 0.030±0.007 UD Daidzein UD UD 0.010±0.001 UD UD means under detection levels Table 2 Effects of dadzein and SDG on the feed efficiency ratio and skeletal muscle growth parameters Control SDG treatment Da treatment Feed efficiency (g g -1 ) 0.225±0.006a 0.253±0.007b 0.271±0.009c Weight of femoral muscle(g) 6.57±0.63a 7.87±0.61b 8.63±0.89b Soleus muscle weight/body weight (g kg -1 ) 0.417±0.034a 0.432±0.027a 0.432±0.029a RNA (mg g -1 ) 0.529±0.041a 0.909±0.062b 1.014±0.044b DNA (mg g -1 ) 0.461±0.080a 0.576±0.122a 0.600±0.068a RNA/DNA 1.147±0.355a 1.578±0.284b 1.692±0.362b P 0.05 Values in the same row that do not share the same superscript letter are significantly different (analysis of variance, P 0.05) 0.05 RNA P 0.05DNA RNA/DNA 37.58% P % P SDG Da 26.24% P % P % P % P * P 0.05 ** P 0.01 Compare with control *: P 0.05, **: P 0.01 Fig. 1 Effects of SDG and daidzein on serum urea-n, testosterone and growth hormones levels in male rats

4 M Mark 1 2 SDG 3 Da * P 0.05 ** P 0.01 M: Marker PUC19 Lanes 1-3 represent the control, SDG, and Da; Compare with control *P 0.05, **P 0.01 Fig. 2 Effect of SDG and daidzein on the expression of ER-β mrna in the soleus muscle and hypothalamus ER-β mrna SDG P 0.05 ER-βmRNA Da P 0.01P 0.05 ER-βmRNA SECO secoisolariciresinol END [15] END ENL [7] Wang [2] SDG SDG END ENL SDG END ENL Kah [16] Bowey [14] Knust [17] SDG END ENL 3 g t -1 RNA /DNA [18] 5 mg kg -1 [19] [20-21] RNA DNA RNA/DNA estradiol E 2 α estrogen receptorα ERα β ERβ ERβ [22-23]

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