兒童腫瘤化療前后胃腸道粘膜屏障功能的測定及其意義
發(fā)布時間：2018-09-08 07:21
【摘要】：目的：二胺氧化酶（Diamine oxidase，DAO）存在于哺乳動物的粘膜或絨毛上層,其中大部分存在于小腸粘膜絨毛,極少部分存在于子宮內(nèi)膜絨毛中；DAO是一種具有高度活性的細胞內(nèi)酶,它的活性高低與絨毛高度及粘膜細胞的核酸、蛋白合成關系密切,是反映小腸粘膜結構及功能的較為理想的指標之一。組織和血漿中DAO含量變化來自于兩條途徑：其一是由于壞死的腸粘膜細胞脫落入腸腔,隨之引起腸粘膜DAO活性的降低；其二則是腸腔內(nèi)的DAO進入腸細胞間隙淋巴管和毛細血管,引起血液中DAO水平升高。因此,在臨床實驗和動物實驗的研究中,我們可以通過測定血液中及小腸組織中的DAO水平變化，來反映小腸的粘膜屏障功能狀態(tài),尤其是能在無創(chuàng)條件下測定血中DAO的活性來反映腸道損傷和修復情況。脂肪酸結合蛋白(Fatty acid binding protein，F(xiàn)ABP)是一組低分子量(15kD左右)結合長鏈脂肪酸的結合蛋白，其組織特異性較強，目前研究認為它存在于哺乳動物的心肌、小腸、肝臟、脂肪組織、腦、表皮等的特定組織部位，是監(jiān)測組織損傷較為新興且價值比較高的生物學指標之一；目前脂肪酸結合蛋白的測定多應用于：慢性心力衰竭(H-FABP)，肝臟移植排斥反應(L-FABP)，非心跳供體腎的存活性選擇(H-FABP、L-FABP)；專業(yè)運動員肌肉損傷的預防(S-FABP)等的研究中。腸脂肪酸結合蛋白(Intestinal fatty acid binding protein，I-FABP)作為目前已發(fā)現(xiàn)的FABP的九種類型之一，它的組織特異性高，因其全部來源于小腸上皮細胞的細胞質(zhì)，主要位于小腸黏膜微絨毛的尖端。目前臨床研究表明：I-FABP可以作為診斷早期腸黏膜損害和缺血的指標。而本實驗通過測定化療前后腫瘤患兒血漿中二胺氧化酶（DAO）及腸脂肪酸結合蛋白（I-FABP）的水平，同時相互比較其水平高低的差異，來研究不同強度化療藥物對腸粘膜屏障功能損傷的程度。 方法：將反復化療的患兒按照化療藥物致吐強度的不同分為3組（輕度致吐組，中度致吐組,高度致吐組），采集各組患兒化療前后不同時相（化療前，化療后第3天，化療后第7天）的血漿，采用酶聯(lián)免疫吸附測試法及分光光度計比色法，對血漿標本做DAO及I-FABP的水平測定；同時記錄患兒化療期間常規(guī)應用止吐藥后仍出現(xiàn)的嘔吐次數(shù)，根據(jù)嘔吐次數(shù)多少分為強嘔吐組（"g3次）及弱嘔吐組（3次）；采集每日新鮮大便標本涂片革蘭氏染色、高倍鏡下讀數(shù)，并計算出桿菌/細菌總數(shù)的比例。 計量資料采用均數(shù)±標準差(X±S)表示，計數(shù)資料采用相對比表示，實驗數(shù)據(jù)用SPSS13.0軟件進行統(tǒng)計學分析。檢驗水平α=0.05，P0.05視為差異有統(tǒng)計學意義。 結果： 1.不同致吐強度的各化療組的DAO水平變化比較： （1）化療前，不同致吐強度組之間的血漿DAO水平基本相同，各組間差異無統(tǒng)計學意義（P0.05）； （2）化療后第3天及化療后第7天，不同致吐強度組間的DAO水平存在顯著性差異（P0.05），進一步分析發(fā)現(xiàn)，其中高致吐組DAO水平明顯高于低、中致吐組，其差異有統(tǒng)計學意義（P0.05）； （3）化療后第3天及化療后第7天，各組血漿DAO水平均顯著高于該組化療前水平，各組前后差異均具有統(tǒng)計學意義（P0.05），進一步分析發(fā)現(xiàn)，化療后第3天各組血漿DAO水平均高于該組化療后第7天的DAO水平，各組差異均具有統(tǒng)計學意義（P0.05）。 2.不同致吐強度的各化療組的I-FABP水平變化比較： （1）化療前，不同致吐強度組之間的血漿I-FABP水平基本相同，各組間差異無統(tǒng)計學意義（P0.05）； （2）化療后第3天及化療后第7天，不同致吐強度組間的I-FABP水平存在顯著性差異（P0.05），進一步分析發(fā)現(xiàn)，，高致吐組I-FABP水平明顯高于低、中致吐組（P0.05），其差異均具有統(tǒng)計學意義（P0.05）； （3）化療后第3天及化療后第7天，各組血漿I-FABP水平均顯著高于該組化療前水平，各組前后差異具有統(tǒng)計學意義（P0.05），進一步分析發(fā)現(xiàn)，各組化療后第3天血漿I-FABP水平均高于該組化療后第7天血漿I-FABP水平，各組之間差異均具有統(tǒng)計學意義（P0.05）。 3.不同致吐強度的各化療組之間的嘔吐頻率比較： （1）將各不同致吐強度組之間嘔吐頻率進行統(tǒng)計學分析，結果顯示：高度組多于其他兩組，而中度組多于低度組，但三組間差異無統(tǒng)計學意義（P0.05）。 （2）高度致吐組在化療后第3天其嘔吐頻率達峰值，而中、低度組則于化療后第4天嘔吐頻率達峰值。 4.嘔吐頻率與血漿DAO及I-FABP的關系： （1）直線相關分析結果顯示，所有化療患兒嘔吐頻率與血漿DAO水平呈直線正相關（r=0.744，P0.05）； （2）直線相關分析結果顯示，所有化療患兒嘔吐頻率與血漿I-FABP水平呈直線正相關（r=0.889，P0.05）； （3）在化療第3天，強嘔吐組的血漿DAO及I-FABP水平均顯著高于弱嘔吐組（P0.05）；在化療第7天，強嘔吐組的血漿DAO及I-FABP水平與弱嘔吐組相比無明顯差異（P0.05）。 5.不同致吐強度的各化療組間的腸道菌群情況的比較： 對各致吐強度組間大便桿菌/細菌總數(shù)進行統(tǒng)計學分析發(fā)現(xiàn)，高度組低于其他兩組，中度組低于低度組，但是三組之間差異無統(tǒng)計學意義（P0.05）。 6.腸道菌群與血漿DAO、I-FABP的關系： （1）直線相關分析結果顯示，化療組患兒的腸道菌群狀況（桿菌/細菌總數(shù)比值）與血漿DAO水平呈負相關（r=-0.180，P0.05）； （2）直線相關分析結果顯示，化療組患兒的腸道菌群狀況（桿菌/細菌總數(shù)比值）與血漿I-FABP水平呈負相關（r=-0.222，P0.05）。 結論： 1.各致吐組血漿DAO及I-FABP水平在化療后第3天及第7天均顯著高于該組化療前水平，其中強嘔吐組血漿DAO及I-FABP水平明顯高于弱嘔吐組，說明化療患兒均存在不同程度胃腸黏膜屏障損傷，且隨著化療藥物致吐強度的增加，胃腸粘膜損傷越嚴重，其引起的血漿DAO及I-FABP水平升高亦越明顯。 2.化療組患兒血漿DAO及I-FABP水平的變化與患兒嘔吐頻率均呈正相關，提示血漿DAO及I-FABP的變化可以反映患兒接受化療后的胃腸道功能狀態(tài)的改變情況。 3.化療組患兒血漿DAO及I-FABP水平的變化與腸道菌群中桿菌/細菌總數(shù)呈負相關，提示化療藥物所導致的胃腸道損傷越重，腸道菌群失衡的表現(xiàn)就越明顯。
[Abstract]:OBJECTIVE: Diamine oxidase (DAO) exists in mammalian mucosa or suprachorillary layer, most of which exist in intestinal mucosal villi, but few in endometrial villi; DAO is a highly active intracellular enzyme, its activity is related to the height of villi and the synthesis of nucleic acid and protein in mucosal cells. The changes of DAO content in tissues and plasma come from two pathways: one is that the necrotic intestinal mucosa cells fall off into the intestinal cavity, which leads to the decrease of DAO activity in the intestinal mucosa; the other is that the DAO in the intestinal cavity enters the lymphatic vessels and capillaries of the intestinal cell space. Therefore, in clinical and animal studies, we can reflect the function of intestinal mucosal barrier by measuring the level of DAO in blood and intestinal tissues, especially the activity of DAO in blood under non-invasive conditions to reflect intestinal injury and repair. Fatty acid binding protein (FABP) is a group of low molecular weight (about 15 kD) binding proteins with long-chain fatty acids. Its tissue specificity is strong. At present, it is believed that FABP exists in mammalian myocardium, small intestine, liver, adipose tissue, brain, epidermis and other specific tissues. It is a relatively new and valuable method to monitor tissue damage. Fatty acid binding protein (FABP) is one of the most important biological indicators. At present, the determination of FABP is mostly used in chronic heart failure (H-FABP), liver transplant rejection (L-FABP), non-heart-beating donor kidney viability selection (H-FABP, L-FABP), prevention of muscle injury (S-FABP) of professional athletes, and so on. TTY acid binding protein (I-FABP), one of the nine types of FABP, has high tissue specificity. It originates from the cytoplasm of small intestinal epithelial cells and is mainly located at the tip of small intestinal microvilli. The aim of this study was to determine the levels of diamine oxidase (DAO) and intestinal fatty acid binding protein (I-FABP) in plasma of children with cancer before and after chemotherapy, and to compare the differences between the levels of DAO and I-FABP.
Methods: The children with repeated chemotherapy were divided into three groups according to the intensity of vomiting induced by chemotherapeutic drugs (mild vomiting group, moderate vomiting group, high vomiting group). The plasma samples were collected at different phases before and after chemotherapy (before chemotherapy, 3 days after chemotherapy, 7 days after chemotherapy). Enzyme-linked immunosorbent assay (ELISA) and spectrophotometry were used to detect the plasma. The levels of DAO and I-FABP were measured, and the vomiting frequency was recorded after routine use of antiemetic drugs during chemotherapy. According to the vomiting frequency, the children were divided into strong vomiting group ("g3 times" and weak vomiting group ("3 times"); fresh stool samples were collected daily for Gram staining, high power microscopic reading, and the total number of bacilli/bacteria was calculated. Proportion.
The measurement data were expressed by mean [standard deviation] (X [S], the counting data were expressed by relative comparison, and the experimental data were statistically analyzed by SPSS 13.0 software.
Result:
1. the levels of DAO in different chemotherapeutic groups with different vomiting intensity were compared.
(1) Before chemotherapy, the levels of plasma DAO in different emetic intensity groups were basically the same, and there was no significant difference between each group (P 0.05).
(2) On the 3rd day after chemotherapy and the 7th day after chemotherapy, there were significant differences in the levels of DAO between the groups with different emetic intensity (P 0.05). Further analysis showed that the level of DAO in the high emetic group was significantly higher than that in the low emetic group, and the difference was statistically significant (P 0.05).
(3) On the 3rd day after chemotherapy and the 7th day after chemotherapy, the levels of plasma DAO in each group were significantly higher than those before chemotherapy, and the differences were statistically significant (P 0.05). Further analysis showed that the levels of plasma DAO in each group on the 3rd day after chemotherapy were higher than those on the 7th day after chemotherapy, and the differences were statistically significant (P 0.05).
2. the levels of I-FABP in different chemotherapeutic groups with different vomiting intensity were compared.
(1) Before chemotherapy, the plasma I-FABP levels of different emetic intensity groups were basically the same, and there was no significant difference between each group (P 0.05).
(2) On the third day after chemotherapy and the seventh day after chemotherapy, there was significant difference in the level of I-FABP between the groups with different emetic intensity (P 0.05). Further analysis showed that the level of I-FABP in the high emetic group was significantly higher than that in the low emetic group (P 0.05).
(3) On the 3rd day after chemotherapy and the 7th day after chemotherapy, the levels of plasma I-FABP in each group were significantly higher than those before chemotherapy, and the difference was statistically significant (P 0.05). Meaning (P0.05).
3. the frequency of vomiting between different chemotherapeutic groups with different vomiting intensity was compared:
(1) The vomiting frequency was statistically analyzed among different intensity groups. The results showed that the vomiting frequency in the high group was more than that in the other two groups, while the vomiting frequency in the middle group was more than that in the low group, but there was no significant difference among the three groups (P 0.05).
(2) The vomiting frequency peaked at the 3rd day after chemotherapy in the highly vomiting group, and peaked at the 4th day after chemotherapy in the middle and low vomiting group.
4. the relationship between the frequency of vomiting and plasma DAO and I-FABP:
(1) Linear correlation analysis showed that vomiting frequency was positively correlated with plasma DAO level (r = 0.744, P 0.05).
(2) Linear correlation analysis showed that vomiting frequency was positively correlated with plasma I-FABP level (r = 0.889, P 0.05).
(3) On the third day of chemotherapy, the plasma DAO and I-FABP levels in the strong vomiting group were significantly higher than those in the weak vomiting group (P 0.05); on the seventh day of chemotherapy, the plasma DAO and I-FABP levels in the strong vomiting group were not significantly different from those in the weak vomiting group (P 0.05).
5. comparison of intestinal microflora among different chemotherapeutic groups with different vomiting intensity:
Statistical analysis of the total fecal bacilli/bacteria counts among the vomiting intensity groups showed that the high group was lower than the other two groups, and the moderate group was lower than the low group, but there was no significant difference among the three groups (P 0.05).
6. the relationship between intestinal flora and plasma DAO and I-FABP:
(1) Linear correlation analysis showed that there was a negative correlation between the intestinal flora status (bacillus / bacteria ratio) and plasma DAO level (r = - 0.180, P 0.05).
(2) Linear correlation analysis showed that there was a negative correlation between the intestinal flora status (Bacillus/Bacterial count ratio) and plasma I-FABP level (r=-0.222, P 0.05).
Conclusion:
1. The plasma levels of DAO and I-FABP in each vomiting group were significantly higher than those before chemotherapy on the 3rd and 7th day after chemotherapy. The plasma levels of DAO and I-FABP in the strong vomiting group were significantly higher than those in the weak vomiting group, indicating that there were different degrees of gastrointestinal mucosal barrier damage in the chemotherapy group, and the more severe the gastrointestinal mucosal damage was with the increase of the intensity of vomiting induced by chemotherapy drugs. The level of plasma DAO and I-FABP increased significantly.
2. The changes of plasma DAO and I-FABP levels were positively correlated with the vomiting frequency, suggesting that the changes of plasma DAO and I-FABP could reflect the changes of gastrointestinal function after chemotherapy.
3. The changes of plasma DAO and I-FABP levels were negatively correlated with the total number of bacteria/bacteria in intestinal flora, suggesting that the more serious the gastrointestinal tract injury caused by chemotherapy drugs, the more obvious the imbalance of intestinal flora.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R730.5
本文編號：2229788
[Abstract]:OBJECTIVE: Diamine oxidase (DAO) exists in mammalian mucosa or suprachorillary layer, most of which exist in intestinal mucosal villi, but few in endometrial villi; DAO is a highly active intracellular enzyme, its activity is related to the height of villi and the synthesis of nucleic acid and protein in mucosal cells. The changes of DAO content in tissues and plasma come from two pathways: one is that the necrotic intestinal mucosa cells fall off into the intestinal cavity, which leads to the decrease of DAO activity in the intestinal mucosa; the other is that the DAO in the intestinal cavity enters the lymphatic vessels and capillaries of the intestinal cell space. Therefore, in clinical and animal studies, we can reflect the function of intestinal mucosal barrier by measuring the level of DAO in blood and intestinal tissues, especially the activity of DAO in blood under non-invasive conditions to reflect intestinal injury and repair. Fatty acid binding protein (FABP) is a group of low molecular weight (about 15 kD) binding proteins with long-chain fatty acids. Its tissue specificity is strong. At present, it is believed that FABP exists in mammalian myocardium, small intestine, liver, adipose tissue, brain, epidermis and other specific tissues. It is a relatively new and valuable method to monitor tissue damage. Fatty acid binding protein (FABP) is one of the most important biological indicators. At present, the determination of FABP is mostly used in chronic heart failure (H-FABP), liver transplant rejection (L-FABP), non-heart-beating donor kidney viability selection (H-FABP, L-FABP), prevention of muscle injury (S-FABP) of professional athletes, and so on. TTY acid binding protein (I-FABP), one of the nine types of FABP, has high tissue specificity. It originates from the cytoplasm of small intestinal epithelial cells and is mainly located at the tip of small intestinal microvilli. The aim of this study was to determine the levels of diamine oxidase (DAO) and intestinal fatty acid binding protein (I-FABP) in plasma of children with cancer before and after chemotherapy, and to compare the differences between the levels of DAO and I-FABP.
Methods: The children with repeated chemotherapy were divided into three groups according to the intensity of vomiting induced by chemotherapeutic drugs (mild vomiting group, moderate vomiting group, high vomiting group). The plasma samples were collected at different phases before and after chemotherapy (before chemotherapy, 3 days after chemotherapy, 7 days after chemotherapy). Enzyme-linked immunosorbent assay (ELISA) and spectrophotometry were used to detect the plasma. The levels of DAO and I-FABP were measured, and the vomiting frequency was recorded after routine use of antiemetic drugs during chemotherapy. According to the vomiting frequency, the children were divided into strong vomiting group ("g3 times" and weak vomiting group ("3 times"); fresh stool samples were collected daily for Gram staining, high power microscopic reading, and the total number of bacilli/bacteria was calculated. Proportion.
The measurement data were expressed by mean [standard deviation] (X [S], the counting data were expressed by relative comparison, and the experimental data were statistically analyzed by SPSS 13.0 software.
Result:
1. the levels of DAO in different chemotherapeutic groups with different vomiting intensity were compared.
(1) Before chemotherapy, the levels of plasma DAO in different emetic intensity groups were basically the same, and there was no significant difference between each group (P 0.05).
(2) On the 3rd day after chemotherapy and the 7th day after chemotherapy, there were significant differences in the levels of DAO between the groups with different emetic intensity (P 0.05). Further analysis showed that the level of DAO in the high emetic group was significantly higher than that in the low emetic group, and the difference was statistically significant (P 0.05).
(3) On the 3rd day after chemotherapy and the 7th day after chemotherapy, the levels of plasma DAO in each group were significantly higher than those before chemotherapy, and the differences were statistically significant (P 0.05). Further analysis showed that the levels of plasma DAO in each group on the 3rd day after chemotherapy were higher than those on the 7th day after chemotherapy, and the differences were statistically significant (P 0.05).
2. the levels of I-FABP in different chemotherapeutic groups with different vomiting intensity were compared.
(1) Before chemotherapy, the plasma I-FABP levels of different emetic intensity groups were basically the same, and there was no significant difference between each group (P 0.05).
(2) On the third day after chemotherapy and the seventh day after chemotherapy, there was significant difference in the level of I-FABP between the groups with different emetic intensity (P 0.05). Further analysis showed that the level of I-FABP in the high emetic group was significantly higher than that in the low emetic group (P 0.05).
(3) On the 3rd day after chemotherapy and the 7th day after chemotherapy, the levels of plasma I-FABP in each group were significantly higher than those before chemotherapy, and the difference was statistically significant (P 0.05). Meaning (P0.05).
3. the frequency of vomiting between different chemotherapeutic groups with different vomiting intensity was compared:
(1) The vomiting frequency was statistically analyzed among different intensity groups. The results showed that the vomiting frequency in the high group was more than that in the other two groups, while the vomiting frequency in the middle group was more than that in the low group, but there was no significant difference among the three groups (P 0.05).
(2) The vomiting frequency peaked at the 3rd day after chemotherapy in the highly vomiting group, and peaked at the 4th day after chemotherapy in the middle and low vomiting group.
4. the relationship between the frequency of vomiting and plasma DAO and I-FABP:
(1) Linear correlation analysis showed that vomiting frequency was positively correlated with plasma DAO level (r = 0.744, P 0.05).
(2) Linear correlation analysis showed that vomiting frequency was positively correlated with plasma I-FABP level (r = 0.889, P 0.05).
(3) On the third day of chemotherapy, the plasma DAO and I-FABP levels in the strong vomiting group were significantly higher than those in the weak vomiting group (P 0.05); on the seventh day of chemotherapy, the plasma DAO and I-FABP levels in the strong vomiting group were not significantly different from those in the weak vomiting group (P 0.05).
5. comparison of intestinal microflora among different chemotherapeutic groups with different vomiting intensity:
Statistical analysis of the total fecal bacilli/bacteria counts among the vomiting intensity groups showed that the high group was lower than the other two groups, and the moderate group was lower than the low group, but there was no significant difference among the three groups (P 0.05).
6. the relationship between intestinal flora and plasma DAO and I-FABP:
(1) Linear correlation analysis showed that there was a negative correlation between the intestinal flora status (bacillus / bacteria ratio) and plasma DAO level (r = - 0.180, P 0.05).
(2) Linear correlation analysis showed that there was a negative correlation between the intestinal flora status (Bacillus/Bacterial count ratio) and plasma I-FABP level (r=-0.222, P 0.05).
Conclusion:
1. The plasma levels of DAO and I-FABP in each vomiting group were significantly higher than those before chemotherapy on the 3rd and 7th day after chemotherapy. The plasma levels of DAO and I-FABP in the strong vomiting group were significantly higher than those in the weak vomiting group, indicating that there were different degrees of gastrointestinal mucosal barrier damage in the chemotherapy group, and the more severe the gastrointestinal mucosal damage was with the increase of the intensity of vomiting induced by chemotherapy drugs. The level of plasma DAO and I-FABP increased significantly.
2. The changes of plasma DAO and I-FABP levels were positively correlated with the vomiting frequency, suggesting that the changes of plasma DAO and I-FABP could reflect the changes of gastrointestinal function after chemotherapy.
3. The changes of plasma DAO and I-FABP levels were negatively correlated with the total number of bacteria/bacteria in intestinal flora, suggesting that the more serious the gastrointestinal tract injury caused by chemotherapy drugs, the more obvious the imbalance of intestinal flora.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R730.5
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