欧拉型藏羊消化系统发育和肌肉H-FABP基因表达规律的研究
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摘要
为了探讨藏羊消化系统发育和肌肉H-FABP基因表达规律,本研究选择分布于青藏高原东部黄河第一大弯处甘肃省玛曲县的欧拉型藏羊为研究对象,将60只公羔分别于0、2、7、14、21、28、42、56、72、84、98和112日龄屠宰,采用大体解剖技术、酶活性分析法、生长曲线拟合方程及real time PCR技术等研究了0~112d(哺乳期)欧拉型藏羊羔羊消化道的发育规律及其瘤胃内主要消化酶活性的发育性变化,羔羊不同生理时期皱胃、小肠内容物及黏膜消化酶、胰腺消化酶活性的变化;羔羊消化器官的生长曲线拟合;H-FABP基因在欧拉羊心脏组织和肌肉组织中的发育性表达,以及与IMF含量的相关性,结果如下:
1欧拉型藏羊消化器官的生长发育及瘤胃内主要消化酶活性的变化
测定了各日龄欧拉型藏羊羔羊体重,肝脏、胰腺重量,复胃及内容物重量,复胃净重,小肠、大肠、盲肠净重、相对重,胃容积及肠长度;测定了瘤胃内容物微生物酶活及各胃室pH。结果表明:(1)欧拉型藏羊羔羊在0~112d的消化道发育迅速,消化道各部位增重明显,容积和长度等指标都显著增加,尤其瘤胃和小肠的增重明显高于其他部位的增重速度;(2)欧拉型藏羊羔羊跟群放牧食入部分牧草可明显刺激瘤胃的发育,70d时,瘤胃pH已接近成年羊水平,并维持在相对恒定的范围内,皱胃pH维持较低水平,呈强酸性;(3)瘤胃微生物酶活性随着日龄增长而提高,其中淀粉酶和木聚糖酶活性增加显著,纤维素酶和果胶酶增加较缓慢,蛋白酶缓慢降低,且纤维素酶活性维持在较低水平,112d时羔羊的消化道及其功能已接近成年水平;(4)在欧拉型藏羊生产实践中,70d断奶较适宜,56d是可接受的最早断奶日龄。
2欧拉型藏羊羔羊消化器官发育的生长曲线拟合分析
采用CurveExpert1.3和SPSS11.5分析软件对不同日龄欧拉型藏羊羔羊各消化器官进行四种曲线拟合。结果表明:欧拉型藏羊早期生长过程中体重、体尺及消化器官的变化符合生物自然生长规律,Richards曲线模型是拟合欧拉型藏羊羔羊体重和胸围早期生长的最优模型,R~2分别为0.9900858和0.9956355;Exponential Association是拟合欧拉型藏羊羔羊体高和体长早期生长过程的最优模型,R~2分别为0.9958635和0.9989817;Gompertz Relation模型是拟合羔羊瘤胃、网胃和瓣胃重量生长曲线的最优模型,R~2分别为0.9969633、0.9990474和0.9991166;4th Degree Polynomial Fit模型是皱胃、十二指肠、回肠和盲肠以及肝脏和胰脏重量变化拟合的最优模型,R~2分别为0.9982634、0.9995719、0.9993717、0.9993425、0.9982556和0.9987633;Logistic模型是空肠和直肠重量变化拟合的最优模型,R~2分别为0.9993071和0.9995853;4th Degree Polynomial Fit模型是羔羊各胃室容积变化拟合的最优模型,瘤胃、网胃、瓣胃和皱胃的R~2分别为0.9998086、0.9960602、0.9979316和0.9843221;4th Degree Polynomial Fit模型是拟合羔羊肠道长度生长曲线的最优模型,十二指肠、空肠、回肠、盲肠、结肠和直肠的R~2分别为0.9989660、0.9986684、0.9962898、0.9986193、0.9972662和0.9982142。
3欧拉型藏羊羔羊消化道pH及主要消化酶发育规律
取羔羊皱胃粘膜、内容物,小肠粘膜、内容物等以及胰腺,测定其pH和消化酶活性。结果表明:(1)随着欧拉型藏羊羔羊日龄增大,皱胃(内容物、不同部位黏膜)pH呈现下降趋势,均呈强酸性。初生羔羊皱胃(内容物、不同部位黏膜)pH均显著高于其他日龄(P<0.05)。羔羊到42d后,皱胃内pH变化基本不大(P>0.05);(2)初生羔羊皱胃凝乳酶比活性较高,随着羔羊日龄的增长,酶比活性在迅速降低,在羔羊42d前,皱胃凝乳酶比活性的变化幅度很大,42d后变化幅度较小。皱胃内容物蛋白水解酶比活性相对最低,羔羊皱胃幽门腺区蛋白水解酶比活性低于胃底中部和皱胃贲门腺区(P<0.05),胃底中部和皱胃贲门腺区蛋白水解酶比活性无显著差异(P>0.05)。42d后皱胃胃底中部黏膜蛋白水解酶比活性基本稳定。初生羔羊有较高的胃前脂酶比活性;(3)羔羊胰腺、小肠pH随日龄变化不大:胰腺均大于7,呈弱碱性。回肠pH高于空肠(P<0. 05),空肠的pH高于十二指肠(P<0. 05),十二指肠最低。羔羊十二指肠段pH呈弱酸性,空肠和回肠pH呈弱碱性;(4)羔羊出生后小肠内就存在较高的胰蛋白酶和糜蛋白酶活性。空肠胰蛋白酶和糜蛋白酶活性最高,回肠次之,十二指肠最低。随着羔羊日龄的增长,小肠胰蛋白酶和糜蛋白酶活性呈现为增加趋势,70d后羔羊小肠内容物胰蛋白酶和糜蛋白酶活性基本保持在相对稳定状态;(5)羔羊小肠不同部位淀粉酶活性不同,空肠段淀粉酶活性显著高于十二指肠和回肠(P<0.05)。随着羔羊日龄的增长,小肠淀粉酶活性持续增加,到84d后小肠各段淀粉酶活性基本处于稳定状态。初生羔羊小肠乳糖酶活性较高,小肠不同部位乳糖酶活性不同,空肠段内容物和粘膜乳糖酶活性最高,十二指肠次之,回肠最低;羔羊小肠粘膜乳糖酶活性显著高于其内容物乳糖酶活性;随着羔羊日龄的增长,小肠乳糖酶活性降低,70d后酶活恒定;(6)不同日龄羔羊小肠不同部位内容物和粘膜脂肪酶活性不同,空肠段脂肪酶活性最高,回肠次之,十二指肠最低,且羔羊小肠内容物脂肪酶活性显著高于其粘膜脂肪酶活性;随着羔羊日龄的增长,小肠脂肪酶活性逐渐升高,到84d后小肠各段粘膜脂肪酶活性基本呈稳定趋势;(7)羔羊胰脏主要消化酶活性随日龄变化不大。
4羔羊肌肉H-FABP基因表达的发育性变化及其对肌内脂肪含量的影响
取2、21、56、84和112日龄欧拉型藏羊公羔(各日龄5只)背最长肌、股二头肌和心脏样品,测定肌内脂肪含量和肌肉H-FABP mRNA表达量。结果表明:(1)随着羔羊日龄的增长,欧拉型臧羊肌肉IMF含量持续上升,不同部位组织IMF含量存在差异;(2)随着羔羊日龄的增长,其肌肉H-FABP mRNA的表达不同,并存在组织差异性,2~21d降低,21d后持续上升,各日龄间差异显著;H-FABP mRNA表达量在21~112d羔羊生长期间与IMF含量呈正相关,表明H-FABP基因在生长发育早期就影响IMF含量,可以作为影响欧拉型臧羊IMF含量的候选基因之一。
In order to explore the development of digestive system and expression rule of H-FABP gene in muscle for Tibetan sheep, Oula Tibet sheep that distributed over Maqu County of Gansu Province where located in east Qinghai-Tibet plateau were used as the study objects. 60 male lambs were slaughtered at 0, 2, 7, 14, 21, 28, 42, 56, 72, 84, 98, and 112 daily age respectively. In the study, main technology included dissection technique, active analysis of enzyme, growth curve model, real time PCR and so forth. The important research contents were developmental changes of digestive system, microbe enzyme activity in rumen, digestive enzyme activity in abomasum and small intestine as well as pancreas, growth curve model for digestive organs, expression of H-FABP gene and its correlation with intramuscular fat (IMF) in muscle. The main results are as follows.
1 Developmental changes of digestive enzyme activity in rumen and digestive organs in Oula Tibetan sheep
The determined index were body weight, body measurements, liver weight, pancreas weight, weight of stomach and its content, net weight and relative weight of small intestine and large intestine and cecum, stomach capacity and intestine length, pH in stomachs and digestive enzyme activity in rumen. The results indicated: first, with age increasing, weight, capacity and length of all parts of digestive system were increasing obviously, especially the weight of rumen and small intestine was quicker than that of other parts. Second, due to grazing pasture activate development of rumen, at the age of 70 days, pH in rumen of Oula sheep reached the level of adult sheep and kept relative stability, in addition, the pH in abomasum was lower and showed strong acidity. Third, with age increasing, Oula Tibetan sheep digestive enzyme activity in rumen increased, in which, activity ofα-amylase and xylanase increased obviously, activity of cellulose and pectinase increased slowly, while, activity of protease decreased slowly, activity of cellulose kept lower level. At the age of 112 days, digestive system and function of Oula sheep reached the level of adult sheep. Fourth, in productive practice of Oula sheep, lambs can be weaned at the age of 70 days, 56 daily age is the earliest weaning time which can be accepted.
2 Fitting analysis of growth curve for digestive organs in Oula Tibetan sheep
Fitting growth curve of different digestive organs in Oula sheep was analyzed in CurveExpert 1.3 and SPSS 11.5 software. The results showed: in early age, body weight, body measurements and digestive organs of Oula sheep accord with the natural law of biological growth. Richards model was the best model fitting early growth of body weight and heart girth of Oula sheep, R~2 was 0.9900858 and 0.9956355 respectively. Exponential Association model was the best model fitting early growth of body height and body length of Oula sheep, R~2 was 0.9958635 and 0.9989817 respectively. Gompertz Relation model was the best model fitting growth of rumen, recticulum and omasum weight of Oula sheep, R~2 was 0.9969633, 0.9990474 and 0.9991166 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of abomasum, duodenum, ileum, cecum, liver and pancreas weight of Oula sheep, R~2 was 0.9982634, 0.9995719, 0.9993717, 0.9993425, 0.9982556 and 0.9987633 respectively. Logistic model was the best model fitting growth of jejunum and restum weight of Oula sheep, R~2 was 0.9993071 and 0.9995853 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of stomachs cubage of Oula sheep, as to R~2 of rumen, recticulum, omasum and abomasum, R~2 was 0.9998086, 0.9960602, 0.9979316 and 0.9843221 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of intestine length of Oula sheep, as to R~2 of duodenum, jejunum, ileum, cecum and rectal, R~2 was 0.9989660, 0.9986684, 0.9962898, 0.9986193, 0.9972662 and 0.9982142 respectively.
3 Change of main digestive enzyme and pH in digestive system of Oula sheep
Mucosa and content of abomasum and small intestine as well as pancreas were sampled to determine their pH and activity of digestive enzymes. The results indicated: first, with age increasing, pH of content and mucosa in abomasum of lambs decreased, appearing strong acidity. At birth, pH of content and mucosa in abomasum were significantly higher than that of other daily age (P<0.05)After d42,pH of content and mucosa in abomasum of lambs almost not changed (P>0.05). Second, chymosin activity in abomasum of lambs was higher at birth.With age increasing, chymosin activity decreased quickly. Before d42 chymosin activity in abomasum of lambs changed to large extent,but after d42 chymosin activity in abomasum of lambs almost not changed. Protease activity in abomasums contents was lower.Protease activity in pyloric gland region of abomasum was lower than that of fundic gland region and cardic gland region(P<0.05) protease activity in fundic gland region and cardic gland region was insignificantly(P>0.05). Afte d42 protease activity of fundic gland region in abomasums mucosa kept stability. Pregastric esterase activity in abomasums contents of lambs was higher at birth. Third, with age increasing, pH of pancreas and small intestine almost not changed. pH of pancreas was above 7, appearing alkalescence. pH of ileum was higher than that of jejunum (P<0. 05), pH of jejunum was higher than that of duodenum (P<0. 05), pH of duodenum was lowest. In addition, pH of duodenum appeared feeble acidity, pH of jejunum and ileum appeared alkalescence. Fourth, trypsinase and chymotrypsin activity in small intestine of lambs were higher at birth.Trypsinase and chymotrypsin activity in jejunum were highest, next in ileum, the lowest in duodenum. With age increasing, trypsinase and chymotrypsin activity in small intestine of lambs increased. After d70, trypsinase and chymotrypsin activity in small intestine of lambs kept relatively stable. Fifth,α-amylase activity in different parts of small intestine was different,α-amylase activity in jejunum was significantly higher than that in duodenum and ileum(P<0.05). With daily age of Oula sheep growing,α-amylase activity in small intestine increased continually, after 84 daily age,α-amylase activity in small intestine kept relative stabilization.Lactase activity in small intestine of lambs were higher at birth.Lactase activity in different parts of small intestine was different, lactase activity in jejunum content and mucous was highest, next was duodenum, and ileum was the lowest. Lactase activity in small intestine mucosa was significantly higher that in content. With age increasing, lactase activity in small intestine of lambs decreased. After d70 ,lactase activity in small intestine of lambs kept relatively stable. Sixth, lipase activity in different parts of content and mucosa in small intestine was different for different daily lambs, lipase activity in jejunum was highest, next was ileum, the lowest was duodenum. Lipase activity in small intestine content was significantly higher than that in mucosa(P<0.05). With age increasing, lipase activity in small intestine increased gradually.After d84 lipase activity in small intestine kept relatively stable. Seventh, with age increasing, digestive enzyme in pancreas almost not changed.
4 Developmental change H-FABP gene expression and its effect on IMF in lamb muscle
Longissimus thoracis, biceps femoris muscle and heart of male lambs at the age of 2, 21, 56, 84 and 112 days were sampled respectively. IMF and H-FABP mRNA expression in muscle were analyzed. The results indicated: first, with age increasing, IMF in muscle was increasing continually, in addition, IMF in different tissue was different; second, with age increaing, H-FABP mRNA expression in muscle changed, H-FABP mRNA expression in different tissue was different. From 2 to 21 daily age, H-FABP mRNA expression in muscle decreased, after d21, H-FABP mRNA expression in muscle was increasing continually, the difference at different daily age was significant. From 21 to 112 daily age, H-FABP mRNA expression in muscle and IMF appeared the positive correlation, so, H-FABP gene affected IMF in early growth and development of lambs, H-FABP gene can be used as the candidate gene controlling IMF in Oula Tibetan sheep.
1欧拉型藏羊消化器官的生长发育及瘤胃内主要消化酶活性的变化
测定了各日龄欧拉型藏羊羔羊体重,肝脏、胰腺重量,复胃及内容物重量,复胃净重,小肠、大肠、盲肠净重、相对重,胃容积及肠长度;测定了瘤胃内容物微生物酶活及各胃室pH。结果表明:(1)欧拉型藏羊羔羊在0~112d的消化道发育迅速,消化道各部位增重明显,容积和长度等指标都显著增加,尤其瘤胃和小肠的增重明显高于其他部位的增重速度;(2)欧拉型藏羊羔羊跟群放牧食入部分牧草可明显刺激瘤胃的发育,70d时,瘤胃pH已接近成年羊水平,并维持在相对恒定的范围内,皱胃pH维持较低水平,呈强酸性;(3)瘤胃微生物酶活性随着日龄增长而提高,其中淀粉酶和木聚糖酶活性增加显著,纤维素酶和果胶酶增加较缓慢,蛋白酶缓慢降低,且纤维素酶活性维持在较低水平,112d时羔羊的消化道及其功能已接近成年水平;(4)在欧拉型藏羊生产实践中,70d断奶较适宜,56d是可接受的最早断奶日龄。
2欧拉型藏羊羔羊消化器官发育的生长曲线拟合分析
采用CurveExpert1.3和SPSS11.5分析软件对不同日龄欧拉型藏羊羔羊各消化器官进行四种曲线拟合。结果表明:欧拉型藏羊早期生长过程中体重、体尺及消化器官的变化符合生物自然生长规律,Richards曲线模型是拟合欧拉型藏羊羔羊体重和胸围早期生长的最优模型,R~2分别为0.9900858和0.9956355;Exponential Association是拟合欧拉型藏羊羔羊体高和体长早期生长过程的最优模型,R~2分别为0.9958635和0.9989817;Gompertz Relation模型是拟合羔羊瘤胃、网胃和瓣胃重量生长曲线的最优模型,R~2分别为0.9969633、0.9990474和0.9991166;4th Degree Polynomial Fit模型是皱胃、十二指肠、回肠和盲肠以及肝脏和胰脏重量变化拟合的最优模型,R~2分别为0.9982634、0.9995719、0.9993717、0.9993425、0.9982556和0.9987633;Logistic模型是空肠和直肠重量变化拟合的最优模型,R~2分别为0.9993071和0.9995853;4th Degree Polynomial Fit模型是羔羊各胃室容积变化拟合的最优模型,瘤胃、网胃、瓣胃和皱胃的R~2分别为0.9998086、0.9960602、0.9979316和0.9843221;4th Degree Polynomial Fit模型是拟合羔羊肠道长度生长曲线的最优模型,十二指肠、空肠、回肠、盲肠、结肠和直肠的R~2分别为0.9989660、0.9986684、0.9962898、0.9986193、0.9972662和0.9982142。
3欧拉型藏羊羔羊消化道pH及主要消化酶发育规律
取羔羊皱胃粘膜、内容物,小肠粘膜、内容物等以及胰腺,测定其pH和消化酶活性。结果表明:(1)随着欧拉型藏羊羔羊日龄增大,皱胃(内容物、不同部位黏膜)pH呈现下降趋势,均呈强酸性。初生羔羊皱胃(内容物、不同部位黏膜)pH均显著高于其他日龄(P<0.05)。羔羊到42d后,皱胃内pH变化基本不大(P>0.05);(2)初生羔羊皱胃凝乳酶比活性较高,随着羔羊日龄的增长,酶比活性在迅速降低,在羔羊42d前,皱胃凝乳酶比活性的变化幅度很大,42d后变化幅度较小。皱胃内容物蛋白水解酶比活性相对最低,羔羊皱胃幽门腺区蛋白水解酶比活性低于胃底中部和皱胃贲门腺区(P<0.05),胃底中部和皱胃贲门腺区蛋白水解酶比活性无显著差异(P>0.05)。42d后皱胃胃底中部黏膜蛋白水解酶比活性基本稳定。初生羔羊有较高的胃前脂酶比活性;(3)羔羊胰腺、小肠pH随日龄变化不大:胰腺均大于7,呈弱碱性。回肠pH高于空肠(P<0. 05),空肠的pH高于十二指肠(P<0. 05),十二指肠最低。羔羊十二指肠段pH呈弱酸性,空肠和回肠pH呈弱碱性;(4)羔羊出生后小肠内就存在较高的胰蛋白酶和糜蛋白酶活性。空肠胰蛋白酶和糜蛋白酶活性最高,回肠次之,十二指肠最低。随着羔羊日龄的增长,小肠胰蛋白酶和糜蛋白酶活性呈现为增加趋势,70d后羔羊小肠内容物胰蛋白酶和糜蛋白酶活性基本保持在相对稳定状态;(5)羔羊小肠不同部位淀粉酶活性不同,空肠段淀粉酶活性显著高于十二指肠和回肠(P<0.05)。随着羔羊日龄的增长,小肠淀粉酶活性持续增加,到84d后小肠各段淀粉酶活性基本处于稳定状态。初生羔羊小肠乳糖酶活性较高,小肠不同部位乳糖酶活性不同,空肠段内容物和粘膜乳糖酶活性最高,十二指肠次之,回肠最低;羔羊小肠粘膜乳糖酶活性显著高于其内容物乳糖酶活性;随着羔羊日龄的增长,小肠乳糖酶活性降低,70d后酶活恒定;(6)不同日龄羔羊小肠不同部位内容物和粘膜脂肪酶活性不同,空肠段脂肪酶活性最高,回肠次之,十二指肠最低,且羔羊小肠内容物脂肪酶活性显著高于其粘膜脂肪酶活性;随着羔羊日龄的增长,小肠脂肪酶活性逐渐升高,到84d后小肠各段粘膜脂肪酶活性基本呈稳定趋势;(7)羔羊胰脏主要消化酶活性随日龄变化不大。
4羔羊肌肉H-FABP基因表达的发育性变化及其对肌内脂肪含量的影响
取2、21、56、84和112日龄欧拉型藏羊公羔(各日龄5只)背最长肌、股二头肌和心脏样品,测定肌内脂肪含量和肌肉H-FABP mRNA表达量。结果表明:(1)随着羔羊日龄的增长,欧拉型臧羊肌肉IMF含量持续上升,不同部位组织IMF含量存在差异;(2)随着羔羊日龄的增长,其肌肉H-FABP mRNA的表达不同,并存在组织差异性,2~21d降低,21d后持续上升,各日龄间差异显著;H-FABP mRNA表达量在21~112d羔羊生长期间与IMF含量呈正相关,表明H-FABP基因在生长发育早期就影响IMF含量,可以作为影响欧拉型臧羊IMF含量的候选基因之一。
In order to explore the development of digestive system and expression rule of H-FABP gene in muscle for Tibetan sheep, Oula Tibet sheep that distributed over Maqu County of Gansu Province where located in east Qinghai-Tibet plateau were used as the study objects. 60 male lambs were slaughtered at 0, 2, 7, 14, 21, 28, 42, 56, 72, 84, 98, and 112 daily age respectively. In the study, main technology included dissection technique, active analysis of enzyme, growth curve model, real time PCR and so forth. The important research contents were developmental changes of digestive system, microbe enzyme activity in rumen, digestive enzyme activity in abomasum and small intestine as well as pancreas, growth curve model for digestive organs, expression of H-FABP gene and its correlation with intramuscular fat (IMF) in muscle. The main results are as follows.
1 Developmental changes of digestive enzyme activity in rumen and digestive organs in Oula Tibetan sheep
The determined index were body weight, body measurements, liver weight, pancreas weight, weight of stomach and its content, net weight and relative weight of small intestine and large intestine and cecum, stomach capacity and intestine length, pH in stomachs and digestive enzyme activity in rumen. The results indicated: first, with age increasing, weight, capacity and length of all parts of digestive system were increasing obviously, especially the weight of rumen and small intestine was quicker than that of other parts. Second, due to grazing pasture activate development of rumen, at the age of 70 days, pH in rumen of Oula sheep reached the level of adult sheep and kept relative stability, in addition, the pH in abomasum was lower and showed strong acidity. Third, with age increasing, Oula Tibetan sheep digestive enzyme activity in rumen increased, in which, activity ofα-amylase and xylanase increased obviously, activity of cellulose and pectinase increased slowly, while, activity of protease decreased slowly, activity of cellulose kept lower level. At the age of 112 days, digestive system and function of Oula sheep reached the level of adult sheep. Fourth, in productive practice of Oula sheep, lambs can be weaned at the age of 70 days, 56 daily age is the earliest weaning time which can be accepted.
2 Fitting analysis of growth curve for digestive organs in Oula Tibetan sheep
Fitting growth curve of different digestive organs in Oula sheep was analyzed in CurveExpert 1.3 and SPSS 11.5 software. The results showed: in early age, body weight, body measurements and digestive organs of Oula sheep accord with the natural law of biological growth. Richards model was the best model fitting early growth of body weight and heart girth of Oula sheep, R~2 was 0.9900858 and 0.9956355 respectively. Exponential Association model was the best model fitting early growth of body height and body length of Oula sheep, R~2 was 0.9958635 and 0.9989817 respectively. Gompertz Relation model was the best model fitting growth of rumen, recticulum and omasum weight of Oula sheep, R~2 was 0.9969633, 0.9990474 and 0.9991166 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of abomasum, duodenum, ileum, cecum, liver and pancreas weight of Oula sheep, R~2 was 0.9982634, 0.9995719, 0.9993717, 0.9993425, 0.9982556 and 0.9987633 respectively. Logistic model was the best model fitting growth of jejunum and restum weight of Oula sheep, R~2 was 0.9993071 and 0.9995853 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of stomachs cubage of Oula sheep, as to R~2 of rumen, recticulum, omasum and abomasum, R~2 was 0.9998086, 0.9960602, 0.9979316 and 0.9843221 respectively. 4th Degree Polynomial Fit model was the best model fitting growth of intestine length of Oula sheep, as to R~2 of duodenum, jejunum, ileum, cecum and rectal, R~2 was 0.9989660, 0.9986684, 0.9962898, 0.9986193, 0.9972662 and 0.9982142 respectively.
3 Change of main digestive enzyme and pH in digestive system of Oula sheep
Mucosa and content of abomasum and small intestine as well as pancreas were sampled to determine their pH and activity of digestive enzymes. The results indicated: first, with age increasing, pH of content and mucosa in abomasum of lambs decreased, appearing strong acidity. At birth, pH of content and mucosa in abomasum were significantly higher than that of other daily age (P<0.05)After d42,pH of content and mucosa in abomasum of lambs almost not changed (P>0.05). Second, chymosin activity in abomasum of lambs was higher at birth.With age increasing, chymosin activity decreased quickly. Before d42 chymosin activity in abomasum of lambs changed to large extent,but after d42 chymosin activity in abomasum of lambs almost not changed. Protease activity in abomasums contents was lower.Protease activity in pyloric gland region of abomasum was lower than that of fundic gland region and cardic gland region(P<0.05) protease activity in fundic gland region and cardic gland region was insignificantly(P>0.05). Afte d42 protease activity of fundic gland region in abomasums mucosa kept stability. Pregastric esterase activity in abomasums contents of lambs was higher at birth. Third, with age increasing, pH of pancreas and small intestine almost not changed. pH of pancreas was above 7, appearing alkalescence. pH of ileum was higher than that of jejunum (P<0. 05), pH of jejunum was higher than that of duodenum (P<0. 05), pH of duodenum was lowest. In addition, pH of duodenum appeared feeble acidity, pH of jejunum and ileum appeared alkalescence. Fourth, trypsinase and chymotrypsin activity in small intestine of lambs were higher at birth.Trypsinase and chymotrypsin activity in jejunum were highest, next in ileum, the lowest in duodenum. With age increasing, trypsinase and chymotrypsin activity in small intestine of lambs increased. After d70, trypsinase and chymotrypsin activity in small intestine of lambs kept relatively stable. Fifth,α-amylase activity in different parts of small intestine was different,α-amylase activity in jejunum was significantly higher than that in duodenum and ileum(P<0.05). With daily age of Oula sheep growing,α-amylase activity in small intestine increased continually, after 84 daily age,α-amylase activity in small intestine kept relative stabilization.Lactase activity in small intestine of lambs were higher at birth.Lactase activity in different parts of small intestine was different, lactase activity in jejunum content and mucous was highest, next was duodenum, and ileum was the lowest. Lactase activity in small intestine mucosa was significantly higher that in content. With age increasing, lactase activity in small intestine of lambs decreased. After d70 ,lactase activity in small intestine of lambs kept relatively stable. Sixth, lipase activity in different parts of content and mucosa in small intestine was different for different daily lambs, lipase activity in jejunum was highest, next was ileum, the lowest was duodenum. Lipase activity in small intestine content was significantly higher than that in mucosa(P<0.05). With age increasing, lipase activity in small intestine increased gradually.After d84 lipase activity in small intestine kept relatively stable. Seventh, with age increasing, digestive enzyme in pancreas almost not changed.
4 Developmental change H-FABP gene expression and its effect on IMF in lamb muscle
Longissimus thoracis, biceps femoris muscle and heart of male lambs at the age of 2, 21, 56, 84 and 112 days were sampled respectively. IMF and H-FABP mRNA expression in muscle were analyzed. The results indicated: first, with age increasing, IMF in muscle was increasing continually, in addition, IMF in different tissue was different; second, with age increaing, H-FABP mRNA expression in muscle changed, H-FABP mRNA expression in different tissue was different. From 2 to 21 daily age, H-FABP mRNA expression in muscle decreased, after d21, H-FABP mRNA expression in muscle was increasing continually, the difference at different daily age was significant. From 21 to 112 daily age, H-FABP mRNA expression in muscle and IMF appeared the positive correlation, so, H-FABP gene affected IMF in early growth and development of lambs, H-FABP gene can be used as the candidate gene controlling IMF in Oula Tibetan sheep.
引文
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