目的 探讨糖尿病脓毒症大鼠肺微血管内皮细胞损伤及一氧化氮系统在其发生机制中的作用.方法 中国医科大学动物实验中心,清洁级Wistar大鼠64只随机(随机数字法)分为A、B、C、D四组,(健康对照组,n=16、糖尿病组,n=16、单纯脓毒症组,n=16、糖尿病发生脓毒症组,n=16).糖尿病模型,一次性腹腔注射链脲左菌素60 mg/kg,48 h后随机测定尾静脉末梢血糖≥16.67 mmol/L,为模型成功,饲养4周后下一步试验.脓毒症模型:一次性尾静脉注射大肠杆菌内毒素10 mg/kg体质量.测定全血Tie-2mRNA表达,测定肺脏组织对伊文思蓝渗透性及肺脏干湿质量比、血清及肺组织NO含量,real-time PCR测定肺组织诱生型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)、DDAH2 mRNA水平.数据采用单因素方差分析,并用LSD法进行组间两两比较,P＜0.05为差异有统计学意义.结果 糖尿病脓毒症组较单纯脓毒症组全血Tie-2mRNA表达增多(19.72±0.70) vs.(3.99-±0.92),P=0.00,肺干湿质量比更为降低(0.19±0.01) vs.(0.22±0.01),P =0.000,肺脏对EBD通透性增加更为严重(3.76±0.77) vs.(1.74±0.24),P=0.000,血清—氧化氮水平增高水平D组低于C组,(123.13 ±4.24) vs.(188.30±5.18),P=0.000.在肺组织中表达,两组均显示出高NO水平(53.62 ±6.70),(23.63±3.92) vs.(10.37±1.29),P=0.00,且D组近2倍高于C组(P=0.00)A、B、C组eNOS表达各组之间差异无统计学意义,D组低于A组,(0.07 ±0.02) vs.(0.38 ±0.05),P=0.017; iNOS表达C、D组均高于A组,(80.23±2.49),(32.48 ±5.37) vs.(1.74 ±0.23),均P=0.00；D组高于C组(P=0.00).DDAH2mRNA水平D及C组均低于A组,(0.49±0.13), (7.26±0.50) vs.(11.96±0.55),P均=0.00；D组低于C组(P=0.00).结论 糖尿病脓毒症机体表现为更为严重内皮细胞损伤,严重NO系统调节失衡是糖尿病脓毒症血管内皮损伤加重的可能机制.
目的 探討糖尿病膿毒癥大鼠肺微血管內皮細胞損傷及一氧化氮繫統在其髮生機製中的作用.方法 中國醫科大學動物實驗中心,清潔級Wistar大鼠64隻隨機(隨機數字法)分為A、B、C、D四組,(健康對照組,n=16、糖尿病組,n=16、單純膿毒癥組,n=16、糖尿病髮生膿毒癥組,n=16).糖尿病模型,一次性腹腔註射鏈脲左菌素60 mg/kg,48 h後隨機測定尾靜脈末梢血糖≥16.67 mmol/L,為模型成功,飼養4週後下一步試驗.膿毒癥模型:一次性尾靜脈註射大腸桿菌內毒素10 mg/kg體質量.測定全血Tie-2mRNA錶達,測定肺髒組織對伊文思藍滲透性及肺髒榦濕質量比、血清及肺組織NO含量,real-time PCR測定肺組織誘生型一氧化氮閤酶(inducible nitric oxide synthase,iNOS)、內皮型一氧化氮閤酶(endothelial nitric oxide synthase,eNOS)、DDAH2 mRNA水平.數據採用單因素方差分析,併用LSD法進行組間兩兩比較,P＜0.05為差異有統計學意義.結果 糖尿病膿毒癥組較單純膿毒癥組全血Tie-2mRNA錶達增多(19.72±0.70) vs.(3.99-±0.92),P=0.00,肺榦濕質量比更為降低(0.19±0.01) vs.(0.22±0.01),P =0.000,肺髒對EBD通透性增加更為嚴重(3.76±0.77) vs.(1.74±0.24),P=0.000,血清—氧化氮水平增高水平D組低于C組,(123.13 ±4.24) vs.(188.30±5.18),P=0.000.在肺組織中錶達,兩組均顯示齣高NO水平(53.62 ±6.70),(23.63±3.92) vs.(10.37±1.29),P=0.00,且D組近2倍高于C組(P=0.00)A、B、C組eNOS錶達各組之間差異無統計學意義,D組低于A組,(0.07 ±0.02) vs.(0.38 ±0.05),P=0.017; iNOS錶達C、D組均高于A組,(80.23±2.49),(32.48 ±5.37) vs.(1.74 ±0.23),均P=0.00；D組高于C組(P=0.00).DDAH2mRNA水平D及C組均低于A組,(0.49±0.13), (7.26±0.50) vs.(11.96±0.55),P均=0.00；D組低于C組(P=0.00).結論 糖尿病膿毒癥機體錶現為更為嚴重內皮細胞損傷,嚴重NO繫統調節失衡是糖尿病膿毒癥血管內皮損傷加重的可能機製.
목적 탐토당뇨병농독증대서폐미혈관내피세포손상급일양화담계통재기발생궤제중적작용.방법 중국의과대학동물실험중심,청길급Wistar대서64지수궤(수궤수자법)분위A、B、C、D사조,(건강대조조,n=16、당뇨병조,n=16、단순농독증조,n=16、당뇨병발생농독증조,n=16).당뇨병모형,일차성복강주사련뇨좌균소60 mg/kg,48 h후수궤측정미정맥말소혈당≥16.67 mmol/L,위모형성공,사양4주후하일보시험.농독증모형:일차성미정맥주사대장간균내독소10 mg/kg체질량.측정전혈Tie-2mRNA표체,측정폐장조직대이문사람삼투성급폐장간습질량비、혈청급폐조직NO함량,real-time PCR측정폐조직유생형일양화담합매(inducible nitric oxide synthase,iNOS)、내피형일양화담합매(endothelial nitric oxide synthase,eNOS)、DDAH2 mRNA수평.수거채용단인소방차분석,병용LSD법진행조간량량비교,P＜0.05위차이유통계학의의.결과 당뇨병농독증조교단순농독증조전혈Tie-2mRNA표체증다(19.72±0.70) vs.(3.99-±0.92),P=0.00,폐간습질량비경위강저(0.19±0.01) vs.(0.22±0.01),P =0.000,폐장대EBD통투성증가경위엄중(3.76±0.77) vs.(1.74±0.24),P=0.000,혈청—양화담수평증고수평D조저우C조,(123.13 ±4.24) vs.(188.30±5.18),P=0.000.재폐조직중표체,량조균현시출고NO수평(53.62 ±6.70),(23.63±3.92) vs.(10.37±1.29),P=0.00,차D조근2배고우C조(P=0.00)A、B、C조eNOS표체각조지간차이무통계학의의,D조저우A조,(0.07 ±0.02) vs.(0.38 ±0.05),P=0.017; iNOS표체C、D조균고우A조,(80.23±2.49),(32.48 ±5.37) vs.(1.74 ±0.23),균P=0.00；D조고우C조(P=0.00).DDAH2mRNA수평D급C조균저우A조,(0.49±0.13), (7.26±0.50) vs.(11.96±0.55),P균=0.00；D조저우C조(P=0.00).결론 당뇨병농독증궤체표현위경위엄중내피세포손상,엄중NO계통조절실형시당뇨병농독증혈관내피손상가중적가능궤제.
Objective To investigate the pulmonary microvascular responsiveness of diabetic animals to sepsis and the potential mechanism of NO system.Methods Sixty-four Wistar rats of clean grade were randomly (random number) divided into 4 groups,namely normal control group (group A,n =16),diabetes group (group B,n =16),sepsis group (group C,n =16),diabetes and sepsis group (group D,n =16).Diabetic mellitus model was made in rats with injection of streptozotocin,STZ (65 mg/kg).Successful model was defined as the blood glucose value≥ 16.67 mmol/L 48 hours after injection of STZ.All animals were fed 4 weeks before initiation of next experiment.The sepsis model was established by intravenous injection of LPS (10 mg/kg) in rats.RT-PCR was used to determine the mRNA expression of Tie-2 in rats'blood.The ratio of dry/wet of lung tissue and the extravasation of Evans blue dye into the lung were detected.Quantitation of NO in lung tissue and serum was measured by using Griess method.RT-PCR was also used for determination of iNOS,eNOS,DDAH2 mRNA expressions in lung tissue.Data were analyzed with ANONA and LSD method for comparison between groups,and P ＜ 0.05 was considered statistically significant.Results Compared with septic group.,the diabetic rats with sepsis group demonstrated higher expression of Tie-2 mRNA in blood (19.72 ± 0.70) vs.(3.99 ± 0.92),P =0.00,lower ratio of dry/wet in lung tissue (0.19 ±0.01) vs.(0.22 ±0.01),P =0.000,higher permeability of Evans blue dye into lung tissue (3.76 ± 0.77) vs.(1.74 ± 0.24),P =0.000.Serum NO level was lower in group D than that in group C (123.13 ±4.24) vs.(188.30 ±5.18),P =0.000,however,NO levels in lung tissue of both group D and group C were higher than that in control group (53.62 ± 6.70),(23.63± 3.92) vs.(10.37 ± 1.29),P =0.00,and NO level in group D was higher in 2 times than that in group C (P =0.00).However,there were no differences in eNOS expression among groups A,B and C,but the difference in eNOS expression was present between group D with lower expression and group A,that lower in group D (0.07 ±0.02) vs.(0.38 ±0.05),P=0.017.Compared with group C,the expression of iNOS was higher in group D (80.23 ±2.49),(32.48±5.37) vs.(1.74±0.23),P=0.00),and the expression of DDAH2 was lower in group D (0.49 ±0.13),(7.26 ±0.50) vs.(11.96 ±0.55).Conclusions Diabetic rats with sepsis enhanced endothelial cell damages.Diabetes deteriorates the regulatory activity of NO system,suggesting the potential mechanism of the worsened damages of EC in diabetic sepsis host.
