EFFECTS OF RECOMBINANT GH ON GROWTH, BLOOD BIOCHEMISTRY, AND BODY COMPOSITION OF JUVENILE DABRY'S STURGEON ACIPENSER DABRYANUS
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摘要: 为研究生长激素（GH）重组蛋白对达氏鲟生长、代谢及鱼体肝组分的影响，构建了GH成熟肽融合原核表达载体pET32a-AdGH，并将其在宿主菌BL21（DE3）中诱导表达，重组蛋白rAdGH以包涵体的形式存在。设置了4个试验组，分别在鲟鱼商品饲料中添加含量为0（对照组）、10（低处理组）、50（中处理组）和100 mg/kg（高处理组）的粉状rAdGH，连续投喂8周。结果表明，低水平rAdGH添加能够显著促进鱼体生长，提高特定生长率，降低饵料系数，提高肝体比和脏体比；进一步的分析表明，低水平rAdGH添加可提高除血糖外所有测定的血液指标数值，反映了机体代谢水平的增强。各浓度rAdGH的添加均能提高鱼体肝和肌肉中蛋白质、水分含量，降低脂肪含量。rAdGH对鱼体生长和代谢的影响可能与其剂量相关，低水平的添加可显著增强鱼体代谢水平，加快肝中蛋白质的沉积与脂肪的消耗，促进鱼体生长；高水平的添加进一步增强其代谢水平，但其促生长的功能下降，部分血液指标异常，肝组分异常，疑似肝损伤。研究探索了鲟鱼重组生长激素的生长调控功能，为重组生长激素在鲟鱼养殖中的应用提供了基础。Abstract: In this study, Dabry's sturgeon (Acipenser dabryanus) mature growth hormone (GH) cDNA sequence was constructed with prokaryotic expression vector pET-32a and transfected into E. coli BL21 (DE3). Recombinant A. dabryanus GH protein (rAdGH) was expressed in E. coli cells as an inclusion body. The purified rAdGH were used as a feed additive to determine the effects of rAdGH on growth, blood biochemistry, and body composition of Dabry's sturgeon for an eight-weeks experiment with four groups including control group (0), low-treated group (10 mg/kg), middle-treated group (50 mg/kg), and high-treated group (100 mg/kg). Results showed that the complementary rAdGH improved weight gain rate, feed efficiency, hepatosmatic index, viscera index and growth performance in the low-treated group. In addition, the low level of rAdGH treatment promoted almost all blood indexes except blood glucose. Suitable level of exogenous GH improved the content of protein and moisture in the liver and muscle, and reduced the fat content in the liver and muscle. In general, its effects on growth and metabolism were dosage-dependent. Low level rAdGH treatment significantly improved the metabolism and growth of fish, and promoted deposition of liver protein and fat consumption. However, a higher dose induced metabolic disease, inhibited growth performance, trigged liver damage and changed components. These results suggest that the recombinant GH would benefit the application of recombinant GH in sturgeon culture.
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表 1 rAdGH对达氏鲟幼鱼生长性能的影响(平均值±标准误)
Table 1 Effects of dietary rAdGH levels on growth performance of juvenile Dabry’s sturgeon (Mean±SE)
参数Parameter 饵料中rAdGH添加量Diet group with additive rAdGH (mg/kg) 0 10 50 100 初重Initial weight (g) 18.63±0.08 18.69±0.04 18.70±0.01 18.73±0.07 末重Final weight (g) 106.48±1.71b 115.46±1.52a 110.7±1.38ab 109.1±0.86b 摄食率FR (%) 2.44±0.09 2.43±0.10 2.47±0.08 2.41±0.08 饵料系数FCR 0.8425±0.08 0.8100±0.08 0.8375±0.07 0.8175±0.06 增重率WGR (%) 472±7.29b 518±8.57a 492±7.21b 483±6.13b 特定生长率SGR (%/d) 2.77±0.02b 2.89±0.02a 2.82±0.02b 2.80±0.02b 存活率SR (%) 99.31±0.69 100±0.00 100±0.00 98.61±1.39 肝体比HSI (%) 3.60±0.001b 3.63±0.002b 3.84±0.001b 4.12±0.002a 脏体比VSI (%) 8.88±0.004b 9.34±0.003ab 9.54±0.002ab 9.89±0.003a 肥满度CF (g/cm3) 0.681±0.010 0.683±0.019 0.722±0.016 0.688±0.013 注: 同一行中数据右上方不同字母代表有显著性差异(P<0.05), 反之无显著性差异(P>0.05)(平均值±标准误), 以下各表同 Note: Date presented are Means±SE, Values in the same row with different superscript letters are significantly different (P<0.05); the same applies below 
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表 2 rAdGH对达氏鲟均重的影响(平均值±标准误)
Table 2 Effects of dietary rAdGH on average weight of Dabry’s sturgeonin different time (Mean±SE)
参数Parameter 饵料中rAdGH添加量Diet group with additive rAdGH (mg/kg) 0 10 50 100 初始均重Initial weight 18.63±0.08 18.69±0.04 18.70±0.01 18.73±0.07 第2周均重The second week 33.58±0.23 35.31±0.23 34.36±1.36 33.51±0.85 第4周均重The fourth week 51.60±0.30b 55.29±1.02a 53.86±0.64a 51.22±0.22b 第6周均重The sixth week 78.44±0.77b 83.70±3.90a 79.56±1.52ab 77.79±1.25b 第8周均重The eighth week 106.48±1.71b 115.46±1.52a 110.7±1.38ab 109.1±0.86b
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表 3 rAdGH对达氏鲟幼鱼血液生化指标的影响(平均值±标准误)
Table 3 Effects of dietary rAdGH on serum biochemical indices of juvenile Dabry’s sturgeon (Mean±SE)
参数Parameter 饵料中rAdGH添加量Diet group with additive rAdGH (mg/kg) 0 10 50 100 甘油三酯TG (mmol/L) 4.63±0.09c 5.63±0.33b 5.30±0.14b 6.68±0.37a 血糖Glu (mmol/L) 4.93±0.07 4.62±0.09 5.10±0.10 5.08±0.08 总蛋白TP (g/L) 14.25±0.50b 14.75±0.96b 16.00±0.58a 17.00±0.82a 白蛋白ALB (g/L) 3.35±0.06c 3.65±0.06b 3.83±0.06a 3.69±0.06b 球蛋白GLB (g/L) 10.50±0.58c 10.75±0.29c 11.75±0.29b 14.00±1.15a ALB/GLB 0.32±0.01a 0.34±0.002a 0.33±0.004a 0.27±0.01b 谷草转氨酶AST (U/L) 201±5.29c 209±4.19c 227±2.50ab 232±6.34a 谷丙转氨酶ALT (U/L) 2.0±0.08c 2.0±0.16c 2.5±0.23b 3.25±0.06a 碱性磷酸酶ALP (U/L) 83±0.96b 100±0.96a 104±2.94a 105±3.40a 总胆固醇TCHO (mmol/L) 1.67±0.04c 2.10±0.04b 2.13±0.05b 2.40±0.07a 高密度脂蛋白HDLC (mmol/L) 1.44±0.03c 1.51±0.02b 1.60±0.04a 1.51±0.03b 低密度脂蛋白LDLC (mmol/L) 0.29±0.01c 0.42±0.01b 0.55±0.02a 0.56±0.02a
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表 4 rAdGH对达氏鲟幼鱼肝、肌肉常规组分的影响(平均值±标准误)
Table 4 Effects of dietary rAdGH on proximate composition of liver and muscle of juvenile Dabry’s sturgeon (Mean±SE)
参数Parameter 饲料中rAdGH添加量Diet group with additive rAdGH (mg/kg) 0 10 50 100 肝Liver 水分Moisture 58.75±0.34c 62.69±1.10ab 65.32±2.34a 62.35±1.36b 粗蛋白Crude protein 19.29±0.37b 20.49±0.99ab 21.17±0.48a 20.67±0.57a 粗脂肪Crude lipid 56.07±0.24b 53.50±0.87c 50.85±0.93d 57.44±0.50a 肌肉Muscle 水分Moisture 80.02±0.83ab 79.38±0.27b 81.03±0.91a 80.61±0.46ab 粗蛋白Crude protein 65.16±1.34b 68.18±1.09a 69.25±0.59a 63.78±0.29b 粗脂肪Crude lipid 18.85±0.46ab 19.30±0.39a 18.49±0.40b 17.70±0.07c
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