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Effect of Extract Compounds from Atractylodes Rhizome, Agastache Rugosa on Small Intestinal Digestion and Absorption in Piglets under High Temperature Stress

Author: SongXiaoZhen
Tutor: WangTian
School: Nanjing Agricultural College
Course: Animal Nutrition and Feed Science
Keywords: Agastache Rugosa Rhizome Atractylodes Extract compounds Hightemperature stress Piglet Intestinal absorption
CLC: S828
Type: PhD thesis
Year: 2008
Downloads: 117
Quote: 0
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Abstract


The heat stress in pig could happen under high temperature and high humidity environment in summer. It results in reduction of gastrointestinal nutrient digestion and absorption and accordingly the decrease of production performance, which bring great losing to the pig fostering. The mammalian small intestine is a central organ which is very sensitive to all stressors, which is also the major site of digesting and absorbing the nutrition. Hydrolases and transport proteins, which rich in the brush-border membrane vesicles (BBMVs) and basement membrane of small intestine, play a critical role on intestinal nutrient digestion and absorption. Agastache Rugosa, Rhizome Atractylodes, and Cortex Phellodendron have been reported to improve intestinal absorption on nutrition. Therefore, the current study was designed to investigate the effect of high temperature stress on growth performance and intestinal absorption of piglets under simulating high temperature and humidity environment in summer, and probe the regulation of extract compound preparation, which was extracted by extract compounds from Agastache Rugosa, Rhizome Atractylodes, Cortex Phellodendron and Gypsum Fibrosum on small intestinal digestion and absorption of heat stressed piglets. And then a model of heat stress in rat IEC-6cells was erected and used to select the effective extracts, which is helpful to illuminate the mechanism of the extract compounds on resisting high temperature stress.So the present studies including five parts as bellows.1. Effects of extract compounds from Atractylodes Rhizome, Agastache Rugosa on growth performance and serum indexes in piglets under high temperature stressThe swine was put into the man-made granary to stimulate the weather in summer to heat stress (40℃,5h/d) in order to study the effect of extracts compound preparation I and II, which was extracted from Rhizome Atractylodes, Agastache Rugosa, Cortex Phellodendron and Gypsum Fibrosum, on the growth performance and serum indexes of swine under heat stress. Total sixty-four2-month-old Chinese experimental piglets (CEMP, agriculture university I series) were blocked according to weight, sex and litter origin, and then randomly allotted to three groups and treated as follows:Normal temperature control group (NTG), High temperature control group (HTG); Extract compound preparation group I (ETGI) and Extracts compound preparation group Ⅱ (ETGII)(n=16per group). On the day1,3,6,10of trial, four piglets were selected from each group, weighed and collected blood separately, and serum glucose levels, MDA levels and SOD, GSH-Px activity were analyzed by commercial kit. The result shows that the high temperature stress decreased ADFI and ADG, induced the decline of serum glucose levels, and increased the activity of SOD and GSH-Px in piglets. Extract Ⅰ improved the ADFI and ADG of heat stressed piglets (P>0.05), and decreased serum MDA contents (P<0.05);extract Ⅱ improved porcine growth performance, increased the activity of SOD and GSH-Px in serum, and induced the decline of MDA contents under high temperature stress (P<0.05). The current results indicated that extract Ⅱ plays a more effective role in the preventive and curative of heat stress in swine compared with that of extract Ⅰ.2. Effects of extract compounds from Atractylodes Rhizome, Agastache Rugosa on small intestinal lipid peroxidation in piglets under high temperature stressTo determine the effect of supplemental extracts compound preparation Ⅰ and Ⅱ on small intestinal lipid peroxidate in piglets under high temperature stress. In each group, four piglets were collected duodenum, jejunum, ileum samples after treatment on the day1,3,6, and10respectively, one part were used to make intestinal electron microscopic section, and others were determined SOD、GSH-PX activity、MDA contents in small intestine by kit. The result showed:(1) Compared with the NTG, jejunum SOD、GSH-Px activity in the HTG were significant decreased on day6and10(p<0.05), MDA content was increased on day1and3(p<0.05), and the activities of SOD and GSH-Px in ileum were decreased remarkably on day10(p<0.05).(2) Compared with the HTG, extract Ⅱ decreased MDA content in jejunum on day1and3, and improved intestinal cell membrane structure (p<0.05). These results indicated that extract Ⅱ plays an important role on improving the antioxidant status and cell membrane structure of small intestine in piglets than extract Ⅰ under high temperature stress.3. Effects of extract compounds from Atractylodes Rhizome, Agastache Rugosa on small intestinal digestion and absorption in piglets under high temperature stressTo study the effects of extracts compound preparation Ⅱ on small intestinal digestion and absorption of piglets during high temperature stress. Forty-eight Chinese experimental piglets with2-month-old were blocked according to weight, sex and litter origin, and then randomly allotted to three groups and treated as follows:Normal temperature control group (NTG); High temperature control group (HTG); The extract compound preparation group (ETG)(n=16per group). In each group, duodenum, jejunum, ileum samples were collected respectively from four piglets on day1,3,6and10of heat stress; and then small intestinal brush border membrane vesicle (BBMV) was prepared. Sucrase, lactase, maltase and AKP activity of BBMV were determined by kit, and at the same time small intestinal xylose absorption of piglets were investigated. The result showed: the highest activity of disaccharidases and AKP was detected in jejunum, followed by duodenum and ileum. Disaccharidases and AKP activity of BBMV in duodenum and jejunum in the HTG were decreased significantly (P<0.05) on day1,3,6comparing to the NTG, then trend to normal level on day10(P>0.05); high temperature treatment induced the decreased of intestinal xylose absorption on day3,6. Extracts compound preparation up-regulated significantly disaccharidase activity in BBMV of small intestine during heat stress, and recovered intestinal absorption to normal level ahead on day6after heat stress.4. Effects of extract compounds from Atractylodes Rhizome, Agastache Rugosa on small intestinal glucose transport correctional carrier expression in piglets under high temperature stressTo study the effect of extracts compound preparation on small intestinal absoption in heat stressed piglets, the activity of Na+/K+-ATPase and expressional levels of sodium-glucose co-transporter1(SGLT1), glucose transporter2(GLUT2) were determined by chemical assay and ELISA kit, and a method of real time PCR was applied to determine SGLT1and GLUT2mRNA expression in small intestine on day6. The result showed that heat stress induced a decrease of the activity of Na+/K+-ATPase in small intestine, and GLUT2contents in duodenal and ileal of the HTG were decreased compared with the NTG (P<0.05). The expression of SGLT1and GLUT2mRNA in small intestine with HTG did not change compared to the NTG (P<0.05). The extract compound preparation increased intestinal Na+/K+-ATPase activity and its gene expression, and improved the SGLT1/GLUT2protein levels in small intestine of heat stressed piglets. Only duodenal and jejunal expression of SGLT1and GLUT2genes expression in the ETG were increased when it compared with the NTG and HTG. However, no difference was showed on SGLT1 and GLUT2mRNA expression of ileum among three groups. These results indicated that extracts compound preparation improved significantly the absorption of glucose in small intestine during heat stress, but the extract affected SGLT1and GLUT2mRNA expression in small intestine of piglets in a spatio-temporal specific manner.5. Effects of Atractylodes Rhizome essential oil, Agastache Rugosa essential oil on small intestinal epithelial cells under high temperature stressWe established the small intestinal epithelial cells model of heat stress by using the rat IEC-6cells, and investigated the effects of heat stress on the intestinal epithelail cells proliferation and glucose absorption as well as the regulation of four extracts which composed the extract compounds Ⅱ. Results showed that high temperature stress treatment (41℃,2h) decreased the proliferation of IEC-6cells. Four extracts altered the cell proliferation in a dose-dependent manner, and Agastache Rugosa essential oil(100μg/mL,200μg/mL), Rhizome Atractylodes essential oil and Gypsum Fibrosum water extract (50μg/mL,100μg/mL,200μg/mL)increased the proliferation of IEC-6cells(P<0.05), but Cortex Phellodendron alkaloid with100μg/mL,50μg/mL,100μg/mL,200μg/mL decreased the proliferation of IEC-6cells under both normal and high temperature(P>0.05) Moreover,100μg/mL Agastache Rugosa essential oil treatment increased glucose absorption and Na+/K+-ATPase activity in small intestinal epithelial cell under both normal and high temperature(37℃and41℃, P<0.05), and up-regulated the expression of SGLT1, GLUT2and Na+/K+-ATPase gene in IEC-6cells under heat stress (41℃, P<0.05) but without changing their expression under normal temperature (37℃, P>0.05). However, Rhizome Atractylodes essential oil with50μg/mL improved glucose absorption and the activity of Na+/K+-ATPase, and up-regulated the expression of SGLT1and Na+/K+-ATPase gene in IEC-6cells under normal temperature(37℃, P<0.05), only SGLT1gene expression in cells were increased by Rhizome Atractylodes essential oil treatment under heat stress (41℃,P<0.05)

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