目的 研究肠道缺血再灌注损伤时肠道黏膜损伤和血浆游离氨基酸的变化.方法 24只健康雄性SD大鼠随机分为3组(每组8只):空白组、假手术组和肠道缺血再灌注(I/R)组.测定所有大鼠空肠和回肠黏膜的厚度和绒毛高度,进行下腔静脉取血,血浆沉淀蛋白质后测定游离氨基酸水平.结果 I/R组与空白组和假手术组比较,空、回肠的黏膜厚度和绒毛高度明显下降[空肠黏膜厚度:(401.50±117.79)、(529.22±54.73)、(499.54±64.48) μm,F=31.869,P=0.000；空肠绒毛高度:(271.37±84.29)、(365.26±46.98)、(349.67±56.11)μm,F=30.472,P=0.000；回肠黏膜厚度:(254.20±43.56)、(324.70±30.56)、(298.26±58.46)μm,F=30.442,P=0.000；回肠绒毛高度:(169.37±37.25)、(221.62±37.26)、(193.25±38.39) μm,F=24.145,P=0.000],假手术组和I/R组的血浆总游离氨基酸(F=5.075,P=0.016)、8种必需氨基酸(F=11.216,P=0.000)、谷氨酰胺(F =4.326,P=0.027)和支链氨基酸(BCAA) (F=10.662,P=0.001)较空白组降低,差异均有统计学意义(P均＜0.05),且I/R组的必需氨基酸和支链氨基酸水平明显低于假手术组(P均＜0.05).芳香族氨基酸在3组中的含量差异无统计学意义(F=0.578,P=0.570).假手术组和I/R组的支链氨基酸/芳香族氨基酸比值显著低于空白组[(2.4±0.6)、(1.9±0.4)、(3.1±0.7),F=5.215,P=0.014],I/R组低于假手术组,但差异无统计学意义(P＞0.05).假手术组和I/R组磷酸乙醇胺(F=5.897,P=0.009)、牛磺酸(F=10.702,P=0.001)、瓜氨酸(F=6.360,P=0.007)、胱氨酸(F=15.344,P=0.000)和磷酸丝氨酸(F =4.878,P=0.018)水平较空白组均有所降低,且I/R组的牛磺酸和胱氨酸与假手术组相比明显减少(P均＜0.05).I/R组血浆鸟氨酸(F =4.961,P=0.017)、精氨酸(F=13.940,P=0.000)浓度低于空白组和假手术组,差异均有统计学意义(P均＜0.05).I/R和假手术组色氨酸(F=5.429,P=0.031)和谷氨酸(F=15.241,P=0.000)浓度较空白组增高,且I/R组谷氨酸浓度明显高于假手术组(P＜0.05).结论 大鼠肠道缺血再灌注损伤引起肠道黏膜损伤和血浆游离氨基酸变化,肠道屏障损伤可能与谷氨酰胺浓度下降、蛋白分解增加相关.
目的 研究腸道缺血再灌註損傷時腸道黏膜損傷和血漿遊離氨基痠的變化.方法 24隻健康雄性SD大鼠隨機分為3組(每組8隻):空白組、假手術組和腸道缺血再灌註(I/R)組.測定所有大鼠空腸和迴腸黏膜的厚度和絨毛高度,進行下腔靜脈取血,血漿沉澱蛋白質後測定遊離氨基痠水平.結果 I/R組與空白組和假手術組比較,空、迴腸的黏膜厚度和絨毛高度明顯下降[空腸黏膜厚度:(401.50±117.79)、(529.22±54.73)、(499.54±64.48) μm,F=31.869,P=0.000；空腸絨毛高度:(271.37±84.29)、(365.26±46.98)、(349.67±56.11)μm,F=30.472,P=0.000；迴腸黏膜厚度:(254.20±43.56)、(324.70±30.56)、(298.26±58.46)μm,F=30.442,P=0.000；迴腸絨毛高度:(169.37±37.25)、(221.62±37.26)、(193.25±38.39) μm,F=24.145,P=0.000],假手術組和I/R組的血漿總遊離氨基痠(F=5.075,P=0.016)、8種必需氨基痠(F=11.216,P=0.000)、穀氨酰胺(F =4.326,P=0.027)和支鏈氨基痠(BCAA) (F=10.662,P=0.001)較空白組降低,差異均有統計學意義(P均＜0.05),且I/R組的必需氨基痠和支鏈氨基痠水平明顯低于假手術組(P均＜0.05).芳香族氨基痠在3組中的含量差異無統計學意義(F=0.578,P=0.570).假手術組和I/R組的支鏈氨基痠/芳香族氨基痠比值顯著低于空白組[(2.4±0.6)、(1.9±0.4)、(3.1±0.7),F=5.215,P=0.014],I/R組低于假手術組,但差異無統計學意義(P＞0.05).假手術組和I/R組燐痠乙醇胺(F=5.897,P=0.009)、牛磺痠(F=10.702,P=0.001)、瓜氨痠(F=6.360,P=0.007)、胱氨痠(F=15.344,P=0.000)和燐痠絲氨痠(F =4.878,P=0.018)水平較空白組均有所降低,且I/R組的牛磺痠和胱氨痠與假手術組相比明顯減少(P均＜0.05).I/R組血漿鳥氨痠(F =4.961,P=0.017)、精氨痠(F=13.940,P=0.000)濃度低于空白組和假手術組,差異均有統計學意義(P均＜0.05).I/R和假手術組色氨痠(F=5.429,P=0.031)和穀氨痠(F=15.241,P=0.000)濃度較空白組增高,且I/R組穀氨痠濃度明顯高于假手術組(P＜0.05).結論 大鼠腸道缺血再灌註損傷引起腸道黏膜損傷和血漿遊離氨基痠變化,腸道屏障損傷可能與穀氨酰胺濃度下降、蛋白分解增加相關.
목적 연구장도결혈재관주손상시장도점막손상화혈장유리안기산적변화.방법 24지건강웅성SD대서수궤분위3조(매조8지):공백조、가수술조화장도결혈재관주(I/R)조.측정소유대서공장화회장점막적후도화융모고도,진행하강정맥취혈,혈장침정단백질후측정유리안기산수평.결과 I/R조여공백조화가수술조비교,공、회장적점막후도화융모고도명현하강[공장점막후도:(401.50±117.79)、(529.22±54.73)、(499.54±64.48) μm,F=31.869,P=0.000；공장융모고도:(271.37±84.29)、(365.26±46.98)、(349.67±56.11)μm,F=30.472,P=0.000；회장점막후도:(254.20±43.56)、(324.70±30.56)、(298.26±58.46)μm,F=30.442,P=0.000；회장융모고도:(169.37±37.25)、(221.62±37.26)、(193.25±38.39) μm,F=24.145,P=0.000],가수술조화I/R조적혈장총유리안기산(F=5.075,P=0.016)、8충필수안기산(F=11.216,P=0.000)、곡안선알(F =4.326,P=0.027)화지련안기산(BCAA) (F=10.662,P=0.001)교공백조강저,차이균유통계학의의(P균＜0.05),차I/R조적필수안기산화지련안기산수평명현저우가수술조(P균＜0.05).방향족안기산재3조중적함량차이무통계학의의(F=0.578,P=0.570).가수술조화I/R조적지련안기산/방향족안기산비치현저저우공백조[(2.4±0.6)、(1.9±0.4)、(3.1±0.7),F=5.215,P=0.014],I/R조저우가수술조,단차이무통계학의의(P＞0.05).가수술조화I/R조린산을순알(F=5.897,P=0.009)、우광산(F=10.702,P=0.001)、과안산(F=6.360,P=0.007)、광안산(F=15.344,P=0.000)화린산사안산(F =4.878,P=0.018)수평교공백조균유소강저,차I/R조적우광산화광안산여가수술조상비명현감소(P균＜0.05).I/R조혈장조안산(F =4.961,P=0.017)、정안산(F=13.940,P=0.000)농도저우공백조화가수술조,차이균유통계학의의(P균＜0.05).I/R화가수술조색안산(F=5.429,P=0.031)화곡안산(F=15.241,P=0.000)농도교공백조증고,차I/R조곡안산농도명현고우가수술조(P＜0.05).결론 대서장도결혈재관주손상인기장도점막손상화혈장유리안기산변화,장도병장손상가능여곡안선알농도하강、단백분해증가상관.
Objective To investigate intestinal mucosal injury and the change of free amino acid levels in plasma with intestinal ischemia/reperfusion.Methods Twenty-four Sprague-Dawley (SD) male rats (SPF grade) were randomly divided into 3 groups with 8 rats in each group:Blank group,Sham group and ischemia/reperfusion (I/R) group.The rats in I/R group were subjected to 60 min ischemia by clamping the superior mesenteric artery (SMA),followed by 120 min repeffusion.All rats were sacrificed with blood withdraw through inferior vena cava.The plasma was precipitated with Sulfosalicylic acid and the supernatant free amino acid levels were measured and the intestinal mucosal thickness and villus length were also assayed.Results In the I/R group the total free amino acids,essential amino acids (EAA),glutamine and branched-chain amino acids (BCCA) were remarkably lower [the total free amino acids:I/R vs Blank vs Sham:(4585.1 326.1) vs (5661.5 ±581.9) vs (5337.9±998.7) μmol/L (F=5.075,P=0.016); EAA:I/Rvs Blank vs Sham:(1401.3 ±183.4) vs (2147.6 ± 265.1) vs (1796.2 ± 440.8) μmol/L (F =1 1.216,P =0.000) ; glutamine:I/R vs Blank vs Sham:(646.1 ± 34.7) vs (895.7 ± 258.8) vs (839.1 ± 163.7) μmol/L (F =4.326,P =0.027) ; BCCA:I/R vs Blank vs Sham:(507.8 ± 119.0) vs (912.2 ± 165.8) vs (671.9 ± 79.8) μmol/L (F =10.662,P =0.001)]and the jejunum and ileum mucosal thickness and villus height were decreased compared to Blank and Sham groups [jejunum mucosal thickness:I/R vs Blank vs Sham:(401.50 ± 117.79) vs (529.22 ±54.73) vs (499.54 ±64.48) μm (F=31.869,P =0.000) ; jejunum villus height:I/R vs Blank vs Sham:(271.37 ± 84.29) vs (365.26 ± 46.98) vs (349.67 ± 56.11) μm (F =30.472,P =0.000) ; ileum mucosal thickness:I/R vs Blank vs Sham:(254.20 ± 43.56) vs (324.70 ± 30.56) vs (298.26 ± 58.46) μm (F =30.442,P =0.000) ; ileum villus height:I/R vs Blank vs Sham:(169.37 ± 37.25) vs (221.62 ± 37.26) vs (193.25 ± 38.39) μm (F =24.145,P =0.000)],and The EAA and BCAA in the I/R group were lower than the Sham group (respectively,P ＜0.05).There was no significant difference in aromatic amino acids (AAA) among the three groups [I/R vs Blank vs Sham:(273.2 ± 37.4) vs (296.8 ± 55.6) vs (281.9 ± 7.3) μmol/L (F =0.578,P =0.570)].The ratio BCAA/AAA in the Sham and I/R groups were significantly lower than the Blank group [(I/R vs Blank vs Sham:(2.4 ±0.6) vs.(1.9 ±0.4) vs (3.1 ±0.7) (F =5.215,P =0.014)],while the I/R group was decreased slightly compared to the Sham group,but the difference was not significant (P ＞ 0.05).The ethanolamine phosphate,taurine,citrulline,cystine,phosphoserine levels were reduced in the Sham and I/R groups compared to the Blank group [ethanolamine phosphate:I/R vs Blank vs Sham:(11.4 ± 1.9) vs (14.3 ± 3.4) vs (10.1±1.7) μmol/L(F=5.897,P=0.009);taurine:I/R vs BlankvsSham:(341.1±36.3) vs(533.2±90.8) vs (439.2±105.4) μmol/L (F=10.702,P=0.001); citrulline:I/R vs Blank vs Sham:(57.7±3.2) vs (73.1 ±16.2) vs (58.1 ±3.8) μmol/L (F=6.360,P =0.007); cystine:I/R vs Blank vs Sham:(20.0 ± 3.6) vs (60.6 ± 24.6) vs (36.3 ± 5.8) μmol/L (F =15.344,P =0.000) ; phosphoserine:I/R vs BlankvsSham:(10.2±1.1) vs (15.8±5.4) vs (11.7 ±3.4) μmol/L (F=4.878,P=0.018)],and the taurine and cystine in I/R groups were significantly decreased than the Sham group (respectively,P ＜ 0.05).The ornithine and arginine were comparatively reduced in I/R in contrast to the Blank and Sham groups [ornithine:I/R vs Blank vs Sham:(81.5 ± 19.0) vs (125.5 ±42.3) vs (114.9 ± 19.5) μmol/L (F =4.961,P =0.017) ;arginine:I/R vs Blank vs Sham:(199.2 ± 8.0) vs (258.9 ± 14.6) vs (248.7 ± 38.4) μmol/L (F =13.940,P =0.000)].The tryptophan and glutamic acid concentrations were increased in the Sham and I/R groups [tryptophan:L/R vs Blank vs Sham:(125.9 ± 12.1) vs (103.1 ± 29.9) vs (128.9 ± 18.5) μmol/L (F =5.429,P =0.031) ; glutamic acid:I/R vs Blank vs Sham:(188.6 ± 29.8) vs (93.6 ± 29.4) vs (125.4 ± 43.8) μmol/L (F =15.241,P =0.000)] and it was lower in the Sham group than the I/R group (P ＜ 0.05).Conclusion Intestinal ischemia-reperfusion can cause intestinal mucosal injury and the change of free amino acid levels in plasma and intestinal barrier damage may be related to the decline glutamine concentration and the increase of protein catabolism.
