雷公藤甲素预处理诱导Tregs细胞减轻小鼠肝脏缺血再灌注损伤的实验研究

Objective To investigate the effect and related mechanism of triptolide pretreatment to prevent from ischemia/reperfusion (I/R) injury in mice liver. Methods Sixty male C57BL/6 mouse were randomized into four groups (15/group): A:sham group with saline , B: sham group with triptolide, C: saline I/R group, D: triptolide I/R group. The mice were pretreated with either saline or triptolide (0. 1 mg/kg/d) through intraperitoneal (ip) injection for one week. The mouse partial liver model of I/R injury was established, and samples were collected at 24 h after the I/R injury. Results Serum ALT and AST levels were significantly decreased and histological damage was significantly alleviated in the triptolide I/R group as compared with the saline I/R group (P＜0.05), the concentration of MDA in the triptolide groups was significantly decreased, while SOD activity was significantly increased compared with that of the saline I/R group (P＜0.05). The percentages of CD4+ CD25+ regulatory T cells (Tregs) cells among CD4+ T cells in groups A, B, C, and D were(7. 55 ± 1.87)%, (12. 59±3. 87)%,(7. 85±1.07)%, and(12. 02±3. 16)% in liver tissue, respectively. The expression levels of Foxp3 mRNA were significantly higher in the triptolide I/R group than those of saline I/R group (P＜0. 05). ELISA showed that triptolide could significantly inhibit the levels of IL-6, IL-Iβ and TNF-αand promoted the level of IL-10 in the serum (P＜0.05). Conclusion Pretreatment with triptolide could effectively prevent from liver I/R injury, which may be related to the induction of Treg cells by triptolide, the increase in the level of IL-10 in serum, and the inhibition of IL-6, IL-1β and TNF-α production in serum.     

热休克蛋白70和血红素加氧酶-1表达在肾缺血后处理减轻肾缺血再灌注损伤中的作用

目的 评价热休克蛋白70(HSP70)和血红素加氧酶-1(HO-1)表达在肾缺血后处理减轻肾缺血再灌注损伤中的作用.方法健康雄性SD大鼠140只,体重250～280 g,采用随机数字表法,将大鼠随机分为4组(n=35):假手术组(S组)仅开腹,游离双侧肾脏,分离双侧肾蒂不夹团;肾缺血再灌注组(I/R组)夹闭双侧肾蒂缺血45 min,恢复灌注;缺血后处理组(IPo组)夹闭双侧肾蒂45 min,再灌注10 s,缺血10 s,反复3次,恢复灌注;HSP抑制剂槲皮黄酮+缺血后处理组(Q+IPo组)缺血前1 h 腹腔注射槲皮黄酮100 mg/kg,余操作同IPo组.于再灌注即刻(T0)、1、3、6、12、24、48 h(T1～6)时各组随机取5只大鼠抽心脏血后取肾,检测肾组织HSP70、HO-1的mRNA和蛋白表达,T3时抽心脏血,测定血清肌酐(Cr)和尿素氮(BUN)浓度、caspase-3 mRNA的表达,TUNNEL法检测肾组织凋亡细胞,计算凋亡指数(AI),光镜下观察肾组织病理学结果.结果 与S组比较,其余组T3时血清Cr和BUN浓度和AJ升高,caspase-3 mRNA表达上调,各时点HSF70、BO-1的mRNA和蛋白表达上调(P＜0.05);与I/R组比较,IPo组T3时血清Cr和BUN浓度和AI降低,caspase-3 mRNA表达下调,T1～5时HSP70、HO-1的mRNA和蛋白表达上调(P＜0.05);与IPo组比较,Q+IPo组T3时血清Cr和BUN浓度和AJ升高,caspase-3mRNA表达上调,T1～5时HSP70、HO-1的mRNA和蛋白表达下调(P＜0.05).IPo组肾组织病理学损伤较I/R组减轻,Q+IPo组肾组织病理学损伤程度与I/R组相似.结论 HSP70和H0-1表达参与了肾缺血后处理减轻肾缺血再灌注损伤的过程.

Abstract：

Objective To evaluate the role of the expression of heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1) in the reduction of renal ischemia-reperfusion (I/R) injury by ischemic postconditioning in tats.Methods One hundred and forty healthy male SD rats weighing 250-280 g were randomized into 4 groups ( n = 35 each) : sham operation group (S group) ; I/R group; ischemic postconditioning group (IPo group); quercetin (an inhibitor of HSP) + ischemic postconditioning group (Q + IPo group). Renal I/R was produced by clamping bilateral renal pedicels for 45 min followed by reperfusion. In group S, bilateral kidneys were only exposed through a midline incision but their- pedicels were not clamped. In IPo and Q + IPo groups, 45 min ischemia was followed by three 10 s episodes of ischemia at 10 s intervals for reperfusion and in addition intraperitoneal quercetin 100 mg/kg was injected at 1 h before ischemia in group Q + IPo. Blood samples from hearts were obtained at 0, 1, 3, 6, 12, 24 and 48 h of reperfusion (T0-6) and the rats were then sacrificed and kidneys removed to detect the expression of HSP70 and HO-1 mRNA and protein in renal tissues. The blood samples obtained at T3 were used to determine serum creatinine (Cr) and urea nitrogen (BUN) concentrations and the expression of caspase-3 mRNA . The apoptosis in the renal tissues was detected using TUNEL and apoptotic index ( AI) was calculated. Microscopic examination was performed with light microscope. Results Compared with group S, the serum Cr and BUN concentrations and AI were significantly increased at T3,the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T0-6 in the other groups (P ＜ 0.05) . Compared with group I/R, the serum Cr and BUN concentrations and AI were significantly decreased at T3, the expression of caspase-3 mRNA was down-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T1-5 in group IPo ( P ＜ 0.05) . Compared with group IPo, the serum Cr and BUN concentrations and AI were significantly increased at T3, the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was down-regulated at T1-5, in group Q + IPo ( P ＜ 0.05) . The microscopic examination showed that the renal I/R injury was significantly attenuated by ischemic postconditioning and the degree of injury in group IPo was similar to that in group I/R. Conclusion The expression of HSP70 and HO-1 is involved in the reduction of renal I/R injury by ischemic postconditioning in rats.     

新型铁螯合剂CHGN2957对大鼠肾缺血再灌注损伤的保护作用

目的 观察新型铁螯合剂CHGN2957对大鼠肾缺血再灌注损伤(I/R)急性期的保护作用.方法 选用雄性SD大鼠90只,随机分成5组(n=18):假手术组、CHGN2957高剂量组、低剂量组、溶剂组和阳性对照组.建成大鼠急性肾I/R模型.药物干预均从手术前2 d开始,直至术后12 d结束.观察大鼠死亡率、体质量变化,测定肾功能和肾组织中超氧化物歧化酶(SOD)活力、谷胱甘肽(GSH)还原酶活力和丙二醛(MDA)含量并进行肾组织病理切片观察.结果 术后第3天CHGN2957溶剂组的体质量(242.1±16.2)g、SOD活力(1.23±0.13)U/mg、GSH还原酶活力(336±15)U/L明显低于其他各组(P＜0.05),而血清尿素氮(62.3±3.1)mmol/L、血清肌酐(310.00±21.02)μmol/L、MDA含量(186.5±16.7)nmol/mg明显高于其他各组(P＜0.05).肾组织病理评分显示CHGN2957溶剂组的肾小管损伤明显重于其他各组(P＜0.05).结论 CHGN2957作为一种新型铁螯合剂,在大鼠急性肾I/R过程中能有效减轻大鼠肾I/R,并对肾起保护作用.

Abstract：

Objective To observe the protecgive effects of CHGN2957 on acute renal ischemia/reperfusion injury (I/R) model in rats. Methods Ninety Sprague-Dawley male rats were randomly divided into five groups ( n = 18 each): sham-operative group, high-dose CHGN2957 group, low-dose CHGN2957 group, CHGN2957 vehicle group and positive group. The administration lasted from two days before operation to twelve days after operation. After seccessful establishment of the model, mortality,weight changes, the renal function, superoxide dismutase ( SOD ) activity, glutathione ( GSH ) reductases activity and malondialdehyde (MDA) content in renal tissue were recorded, and the pathological changes were observed. Results Weight (242. 1 ± 16. 2) g, SOD activity (1. 23 ±0. 13) U/mg and GSH reductases activity (336 ± 15 U/L) in CHGN2957 vehicle group were reduced significantly as compared with other groups three days after I/R (P ＜0. 05), but SUN (62. 3 ± 3. 1) mmol/L, SCr (310. 00 ± 21. 02)μmol/L, MDA content ( 186. 5 ± 16. 7 ) nmol/mg were higher than others obviously ( P ＜ 0. 05 ). Pathological observation showed renal tubular injury was more serious in CHGN2957 vehicle group (P＜0.05).Conclusion CHGN2957 as a new iron chelator was effective to ameliorate renal I/R in rats.     

七氟醚预处理对大鼠肾缺血再灌注损伤的影响

目的 评价七氟醚预处理对大鼠肾缺血再灌注损伤的影响.方法 雄性SD大鼠24只,体重250～300 g,采用随机数字表法,将大鼠随机分为3组(n=8):假手术组(S组)、肾缺血再灌注组(I/R组)和七氟醚预处理组(SP组).I/R组和SP组采用切除右肾然后夹闭左侧肾动脉45 min再开放的方法 制备肾缺血再灌注模型.SP组吸入2.2%七氟醚1 h,停止吸入后10 min时进行肾缺血.于再灌注2 h时采集静脉血样,测定血清肌酐(Cr)、尿素氮(BUN)和胱抑素C(Cys C)的浓度,取肾组织,光镜下及透射电镜下观察病理学结果,并根据肾小管病变程度进行Paller评分.结果 与S组比较,I/R组血清Cr和BUN浓度差异无统计学意义(P＞0.05),血清Cys C浓度和Paller评分明显升高(P＜0.05);与I/R组比较,SP组血清Cys C浓度和Paller评分明显降低(P＜0.05).SP组肾组织损伤程度轻于I/R组.结论 七氟醚预处理可减轻大鼠肾缺血再灌注损伤.

Abstract：

Objective To investigate the effects of sevoflurane preconditioning on renal ischemia-reperfusion(I/R)injury in rats.Methods Twenty-four adult male SD rats weighing 250-300 g were randomly divided into 3 groups(n=8 each):sham operation group (group S);I/R group; sevoflurane preconditioning group (group SP). After the rats underwent right nephrectomy, renal I/R was produced by occlusion of left renal artery for 45 min followed by reperfusion in I/R and SP groups.In group SP, the rats inhaled 2.2% sevoflurane for 1 h, then the inhalation was stopped and renal ischemia was performed 10 min later. Venous blood samples were collected at 2 h of reperfusion to determine the concentrations of serum creatinine(Cr), urea nitrogen (BUN), cystatin C (Cys C) . The renal tissues were obtained for microscopic examination, and Paller's score was recorded. Results Compared with group S, there was no significant difference in the serum Cr and BUN concentrations (P＞0.05), while the serum Cys C concentration and Paller's score for acute renal tubular injury were significantly increased in group I/R(P＜0.05). The serum Cys C concentration and Paller's score were significantly lower in group SP than in group I/R(P＜0.05).I/R-induced renal injury was significantly reduced in group SP compared with group I/R. Conclusion Preconditioning with sevoflurane can provide significant protection against renal I/R injury.     

雷公藤甲素预处理诱导Tregs细胞减轻小鼠肝脏缺血再灌注损伤的实验研究

目的 探讨雷公藤甲素(TPT)预处理减轻小鼠肝脏缺血再灌注损伤的作用及其机制.方法 将C57BL/6小鼠随机分为4组(15只/组):假手术对照组;假手术雷公藤甲素组;实验对照组;雷公藤甲素实验组.两个雷公藤甲素组小鼠术前1周每天给予雷公藤甲素0.1 mg/kg腹腔注射,术前1h加用一次,而两对照组同期仅给予等体积无菌生理盐水腹腔注射.肝脏缺血90 min再灌注24 h后分别采集各组小鼠的血液和肝组织,检测血清丙氨酸转移酶(ALT)、天冬氨酸转移酶(AST)水平.以及肝脏组织丙二醛(MDA)和超氧化物歧化酶(SOD)含量.光镜下观察肝组织的病理学变化.采用流式细胞术检测肝组织中CD4+CD25+Foxp3+T淋巴细胞占CD4+T细胞的百分比,采用实时聚合酶链反应检测肝组织中Foxp3 mRNA的表达.采用酶联免疫吸附试验检测血清中IL-10、IL-6、IL-1β和TNF-α细胞因子含量.结果 雷公藤甲素实验组和实验对照组小鼠血清ALT和AST明显升高,且雷公藤甲素实验组水平低于实验对照组(P＜0.05).假手术雷公藤甲素组和雷公藤甲素实验组与相应假手术和实验对照组相比MDA含量降低,SOD活性升高(P＜0.05).两假手术组肝组织结构正常,实验对照组可见明显的肝组织片状坏死,雷公藤甲素实验组肝小叶结构基本正常.假手术对照组、假手术雷公藤甲素组、实验对照组和雷公藤甲素实验组CD4+CD25+Foxp3+T淋巴细胞占CD4+T细胞的百分比分别为(7.55±1.87)%、(12.59±3.87)%、(7.85±1.07)%和(12.02±3.16)%.假手术雷公藤甲素组和雷公藤甲素实验组Foxp3 mRNA相对表达量高于相应的对照组(P＜0.05).雷公藤甲素实验组较实验对照组相比有升高血清IL-10,降低IL-6、IL-1β和TNF-α细胞因子的含量的作用.结论 雷公藤甲素可减轻小鼠肝脏缺血再灌注损伤,其机制可能与雷公藤甲素诱导上调体内CD4+CD25+Foxp3+调节性T淋巴细胞比例及增加IL-10分泌,抑制IL-6、IL-1p和TNF-α炎症细胞因子有关.

Abstract：

Objective To investigate the effect and related mechanism of triptolide pretreatment to prevent from ischemia/reperfusion (I/R) injury in mice liver. Methods Sixty male C57BL/6 mouse were randomized into four groups (15/group): A:sham group with saline , B: sham group with triptolide, C: saline I/R group, D: triptolide I/R group. The mice were pretreated with either saline or triptolide (0. 1 mg/kg/d) through intraperitoneal (ip) injection for one week. The mouse partial liver model of I/R injury was established, and samples were collected at 24 h after the I/R injury. Results Serum ALT and AST levels were significantly decreased and histological damage was significantly alleviated in the triptolide I/R group as compared with the saline I/R group (P＜0.05), the concentration of MDA in the triptolide groups was significantly decreased, while SOD activity was significantly increased compared with that of the saline I/R group (P＜0.05). The percentages of CD4+ CD25+ regulatory T cells (Tregs) cells among CD4+ T cells in groups A, B, C, and D were(7. 55 ± 1.87)%, (12. 59±3. 87)%,(7. 85±1.07)%, and(12. 02±3. 16)% in liver tissue, respectively. The expression levels of Foxp3 mRNA were significantly higher in the triptolide I/R group than those of saline I/R group (P＜0. 05). ELISA showed that triptolide could significantly inhibit the levels of IL-6, IL-Iβ and TNF-αand promoted the level of IL-10 in the serum (P＜0.05). Conclusion Pretreatment with triptolide could effectively prevent from liver I/R injury, which may be related to the induction of Treg cells by triptolide, the increase in the level of IL-10 in serum, and the inhibition of IL-6, IL-1β and TNF-α production in serum.     

ω-3多不饱和脂肪酸和淋巴引流对大鼠肠道缺血再灌注损伤时远隔器官的保护作用

目的 研究大鼠肠道缺血再灌注损伤时ω-3多不饱和脂肪酸干预和肠淋巴引流对远隔组织器官的影响.方法 将48只健康雄性SPF级SD大鼠随机分为正常饮食、普通肠内营养(EN)、普通肠内营养加ω-3多不饱和脂肪酸(PUFA)三大组,每组又分为肠淋巴引流组(I/R+D)和非引流组(I/R)(各8只).所有大鼠均行胃造口手术,分别给予不同营养5 d后行肠系膜上动脉夹闭60 min再灌注120 min.引流组在肠道缺血再灌注同时,进行肠淋巴液引流180 min.检测大鼠血清ALT、肺脏中髓过氧化物酶(MPO)、一氧化氮(NO)、总一氧化氮合酶(tNOS)、诱导型一氧化氮合酶(iNOS)的变化,观察肝脏,肺脏损伤程度以及Toll样受体4(TLR4)的内源性配体高迁移率族蛋白1(HMGB1)的表达.结果 PUFA I/R+D组和I/R组、EN I/R+D组血清ALT水平显著低于正常饮食I/R组,分别为(46±20)、(53±15)、(45±21)和(100±60)U/L(P＜0.05).肺脏MPO、NO、tNOS、iNOS在I/R+D组低于不引流I/R组(P＜0.05),分别为MPO(0.73±0.15)U/g湿片比(0.85±0.10)U/g湿片、NO(0.72±0.51)μmol/gprot比(1.79±1.32)μmol/gprot、tNOS(0.46±0.15)U/mgprot比(0.78±0.27)U/mgprot、iNOS(0.06±0.04)U/mgprot比(0.11±0.07)U/mgprot;PUFA I/R组tNOS显著低于正常饮食I/R组,分别为(0.56±0.13)和(0.78±0.27)U/mgprot(P＜0.05);PUFA组中MPO、NO、iNOS均小于EN和正常饮食组.HE染色以及免疫组化显示I/R组肺和肝组织均较I/R+D组损伤严重,I/R组细胞出现大量黄染,HMGB1的表达增加;PUFA组较另外两组损伤减轻,HMGB1的表达减少.结论 大鼠肠道缺血在灌注损伤时引流淋巴液能够减少HMGB1到达远隔器官组织从而减轻损伤,ω-3PUFA具有增加机体抗打击和促进恢复的能力.

Abstract：

Objective To investigate the sheltering effects of ω-3 polyunsaturated fatty acid (ω-3PUFA)and lymphatic drainage on distant organs in intestinal ischemia-reperfusion injury in rats.Methods Forty-eight healthy Sprague-Dawley(SD)male rats(SPF grade)were randomly divided into 3 groups(16 rats in each group): normal diet group(N), enteral nutrition group(EN), enteral nutrition and ω-3PUFA group(PUFA group). Each group was divided into lymphatic drainage(I/R + D)group and no-drainage(I/R)group(n = 8). Each rats received gastrostomy. After given different nutrition for five days, the rats subjected to 60 min ischemia and 120 min reperfusion injury of the superior mesenteric artery.When the rats subjected to ischemia-reperfusion injury, drained intestinal lymph for 180 min in the I/R + D group. The serum level of alanine aminotransferase(ALT)and level of myeloperoxidase(MPO), nitric oxide(NO), total of nitric oxide synthase(tNOS), inducible nitric oxide synthase(iNOS)of lung were detected. The organ injury of lung and liver and the expression of high mobility group box 1(HMGB1, the endogenous ligand of TLR4)in these organs were investigated too. Results The serum level of ALT in PUFA I/R + D and I/R group and EN I/R + D group were significantly lower than that in normal diet I/R group:(46 ±20),(53 ± 15),(46 ±21)and(100 ±60)U/L(P ＜0. 05), respectively. The level of MPO, NO, tNOS, iNOS in lung in the I/R + D group were significantly lower than those in I/R group(P ＜0.05):MPO(0.73 ±0. 15):(0.85 ±0. 10)unit/grams wet slice; NO(0.72 ±0.51):(1.79 ± 1.32)μmol/gprot; tNOS(0.46 ±0. 15):(0.78 ±0.27)U/mgprot; iNOS(0.06 ±0.04):(0. 11 ±0.07)U/mgprot, respectively. The level of tNOS in PUFA I/R group was significantly lower than that in normal diet I/R group:(0. 56 ±0. 13):(0. 78 ±0. 27)U/mgprot(P ＜0. 05). MPO, NO, INOS levels in PUFA group were reduced compared with those in EN and normal diet group. HE stained sections and HMGB1 immunohistochemistry results showed that the organ injury in L/R group was severer than that in I/R + D group. The expression of HMGB1 increased in I/R group. The organ injury and the expression of HMGB1 in PUFA group were less than that in the other two main groups. Conclusions Lymphatic drainage can alleviate injury of distant organs after intestinal ischemia-reperfusion in rats. ω-3 polyunsaturated fatty acids can increase body resistance to injury and promote recovery.     

硼替佐米对小鼠急性移植物抗宿主病的抑制作用

目的 探讨硼替佐米对小鼠急性移植物抗宿主病(aGVHD)的抑制作用及其可能机制.方法 以C57BL/6小鼠为供鼠,获取骨髓细胞及脾细胞.以Balb/c小鼠为受鼠,共分为5组:空白对照组(n=17)小鼠不予任何处理;单纯照射组(n=17)小鼠仅接受7.0 Gy X射线全身照射(TBI);药物对照组(n=17)小鼠也接受TBI,并且由尾静脉注射硼替佐米;aGVHD组(n=8)小鼠TBI后注射供鼠骨髓细胞及脾细胞;实验组(n=8)小鼠TBI后输注供鼠骨髓细胞及脾细胞,并予以硼替佐米.观察各组小鼠aGVHD的发生情况、存活时间及嵌合状态,蛋白印迹法测定空白对照组、单纯照射组和药物对照组小鼠肝脏及小肠组织细胞核中核因子κB(NF-κB)p65的表达.结果 aGVHD组和实验组的临床aGVHD评分分别为7.37±0.32和5.85±0.40,实验组明显低于aGVHD组(P＜0.05).aGVHD组受鼠肝脏、小肠及皮肤组织为Thomas GVHD病理分级Ⅲ～Ⅳ级改变.实验组受鼠肝脏、小肠及皮肤组织为Ⅰ～Ⅲ级GVHD改变,较aGVHD组有所减轻.实验组存活时间长于aGVHD组(P＜0.05).aGVHD组和实验组小鼠移植后12 d时外周血细胞中H-2Kb分子阳性细胞的百分率均＞90%.药物对照组肝脏及小肠组织细胞核内NF-κB p65表达均高于单纯照射组(P＜0.05).结论 硼替佐米可能通过在一定程度上抑制照射预处理损伤所致肝脏及小肠组织中NF-κB的激活起到减轻aGVHD的作用.

Abstract：

Objective To observe the effect of bortezomib on acute graft-versus-host disease (aGVHD) in an aGVHD model of mice and investigate the related mechanism. Methods Male C57BL/6( H-2Kb)mice were used as donors and female Balb/c (H-2Kd) mice used as recipients. Balb/c mice received total body irradiation (TBI) by 7.0 Gy X-radiation, and randomly divided into five groups. normal (group A), TBI (group B), TBI + bortezomib (group C), TBI + bone marrow cells (BMC) + spleen cells (SC) (group D) and TBI + bortezomib + BMC + SC (group E). The physical signs and the pathological damage of aGVHD, mean survival time, and chimerism were observed in recipients. The NF-κB p65 levels in nuclei of the liver and small intestine tissues of groups A,B and C were analyzed by Western blot. Results ( 1 ) The clinical aGVHD score in group D was (7.37±0. 32), significantly higher than in group E (5.85 ± 0.40) (P＜0. 05). Histopathology of the gut, liver and skin illuminated that the Ⅲ-Ⅳ degree GVHD occurred in group D. The occurrence of aGVHD in group E was later than in group D. The symptoms and the pathological damage of aGVHD in group E were milder than in group D. The average survival time in group E was significantly longer than that in group D (P＜0.05). The percentage of donor-derived cells in recipient mice was above 90% at day 12 after transplantation; (2) NF-κB p65 levels in nuclei of the liver and small intestine tissues in group B was significantly higher than in group C on the day 1,3 and 5 (P＜0. 05). Conclusion Bortezomib can inhibit the activation and expression of NF-κB,which may be the underlying mechanism for it to relieve aGVHD.     

线粒体ATP敏感性钾通道阻断剂对在体大鼠肺缺血后处理的作用及其机制

目的 探讨缺血后处理(IPO)对大鼠在体肺缺血-再灌注损伤(I/R)的保护作用及线粒体ATP敏感性钾通道(mitoKATP)在缺血后处理效应中的作用.方法 将Wistar大鼠35只随机分为5组:假手术组(Sham组)、缺血再灌注损伤组(I/R组)、缺血后处理组(IPO组)、缺血再灌注损伤+5-羟基葵酸盐组(I/R+5-HD组)、缺血后处理+5-羟基葵酸盐组(IPO+5-HD组).观察各组肺组织中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、湿/干比值(W/D)以及病理形态学改变.结果 I/R组与Sham组比较MDA含量增加[(5.07±1.60)nmol/mg prot比(1.43±0.41)nmol/mgprot,P＜0.01],SOD活性减低[(12.38±2.24)U/mg prot比(45.51±5.42)U/mg prot,P＜0.01],W/D比值增高(5.45±0.82比3.05±0.47,P＜0.01),肺组织形态及超微结构明显受损;IPO+5-HD组与IPO组比较MDA含量增加[(3.74±0.71)nmol/mg prot比(2.60±0.43)nmol/mg prot,P＜0.01],SOD活性减低[(22.91±2.71)U/mg prot比(28.74±2.03)U/mg prot,P＜0.01],W/D比值增高(4.64±0.79比3.89±0.60,P＜0.01),肺组织形态及超微结构明显受损;IPO组与I/R组比较,肺组织MDA含量减少[(2.60±0.43)nmol/mg prot比(5.07±1.60)nmol/mg prot,P＜0.01],SOD活性增高[(28.74±2.03)U/mg prot比(12.38±2.24)U/mg prot,P＜0.01],W/D比值减低(3.89±0.60比5.45±0.82,P＜0.01),肺组织病理形态学改变轻于I/R组;I/R+5-HD组与I/R组比较,肺组织MDA含量[(5.14±1.30)mol/mg prot比(5.07±1.60)mol/mg prot,P＞0.05)、SOD活性[(11.65±1.82)U/mg prot比(12.38±2.24)U/mg prot,P＞0.05]、W/D比变化(5.54±0.61比5.45±0.82),差异无统计学意义(P＞0.05),肺组织病理形态学改变无明显差异.IPO+5-HD组的各项指标介于IPO组和I/R组之间.结论 缺血后处理能减轻大鼠在体肺缺血再灌注损伤,mitoKATP参与了肺缺血后处理效应.

Abstract：

Objective To investigate the protective effect of ischemic postconditioning (IPO) on lung ischemic reperfusion (L/R) in rats in vivo and the mechanism of mitochondrial ATP-sensitive potassium channel (mitoKATP) blocker in the ischemic postconditioning. Methods Thirty five Wistar rats were randomly divided into 5 groups: sham group, I/R group, ischemic postconditioning (IPO) group, I/R +5-hydroxydecanoate (I/R + 5-HD) group, IPO + 5-HD group. The concentration of malondialdehyde (MDA) and activity of superoide dismutase (SOD) were determined in the lung homogenate, wet to dry weight ratio (W/D) was measured and pathological changes were also observed. Results The levels of MDA[(5.07±1.60) vs (1.43 ±0.41) nmol/mg prot,P＜0. 01]and W/D (5.45 ±0.82 vs 3.05 ±0. 47,P ＜0. 01 ) were increased significantly in I/R group as compared with sham group, while the activity of SOD[( 12. 38 ±2. 24) vs (45.51 ±5.42) U/mg prot,P ＜0. 01]was decreased, and the injury of lung tissues was significantly aggravated in IPO + 5-HD group as compared with IPO group[MDA: (3.74 ±0. 71 ) nmol/mg prot vs (2. 60 ± 0. 43 ) nmol/mg prot , P ＜ 0. 01]; W/D: 4. 64 ± 0. 79 vs 3. 89 ± 0. 60,P＜0.01; SOD:[(22.91 ±2.71) U/mg prot vs (28.74±2.03) U/mg prot,P＜0. 01]. The levels of MDA[(2.60±0.43) vs (5.07 ±1.60) nmol/mg prot,P＜0. 01]and W/D (3.89 ±0.60 vs 5.45 ±0. 82,P ＜0. 01 ) were decreased significantly in IPO group as compared with I/R group, the activity of SOD[(28.74±2.03) vs (12.38 ±2.24) U/mg prot,P＜0. 01]increased and lung tissue histological damage attenuated. The difference in MDA[(5.14 ± 1.30) vs (5.07 ± 1.60) nmol/mg prot, P ＞ 0. 05],W/D (5.54±0.61 vs5.45 ±0.82,P＞0.05) and SOD[(11.65 ±1.82) vs (12.38 ±2.24) U/mgprot,P ＞ 0. 05]levels had no statistical significance between I/R + 5-HD group and I/R group, and the injury of lung tissues had no significant difference too. Each index in IPO + 5-HD group was between IPO and I/R groups. Conclusion Ischemic postconditioning can attenuate the lung I/R injury, and mitoKATP plays a vital role in the protective procession of ischemic postconditioning on lung ischemic reperfusion.     

不同剂量瑞芬太尼对大鼠肾脏缺血再灌注损伤的影响

目的 探讨不同剂量瑞芬太尼对大鼠肾脏缺血再灌注损伤的影响.方法 健康成年雄性SD大鼠60只,体重220～250 g,采用随机数字表法,将大鼠随机分为5组(n=12):假手术组(S组)、模型组(M组)、低、中和高剂量瑞芬太尼组(RL组、RM组和RH组).除S组外均采用夹闭双侧肾动脉45 min后恢复再灌注法建立肾脏缺血再灌注模型.RL组、RM组和RH组于缺血前15min分别经尾静脉输注瑞芬太尼0.2、0.6、1.0μg·kg-1·min-1至再灌注30 min;S组和M组给予等容量生理盐水替代.于再灌注30 min及24 h时经股静脉采集血样1 ml,测定血清BUN及Cr浓度;于再灌注24h时取肾组织,测定MDA含量及SOD、Ca2+-ATP酶活性,光、电镜下观察肾组织病理学结果.结果 与S组比较,其余4组血清BUN和Cr浓度、肾组织MDA含量升高,SOD和Ca2+-ATP酶活性降低(P＜0.05或0.01),肾组织有不同程度的病理学损伤.与M组比较,RL组、RM组和RH组血清BUN和Cr浓度、肾组织MDA含量降低,SOD和Ca2+-ATP酶活性升高(P＜0.05或0.01),肾组织病理学损伤减轻o RL组、RM组和RH组随瑞芬太尼剂量增加,血清BUN和Cr浓度、肾组织MDA含量逐渐降低,SOD和Ca2+-ATP酶活性逐渐升高(P＜0.05或0.01),肾组织病理学损伤逐渐减轻.结论 瑞芬太尼可减轻大鼠肾脏缺血再灌注损伤,且与剂量有关,其机制与抑制脂质过氧化反应、提高Ca2+-ATP酶活性有关.

Abstract：

Objective To investigate the effects of different doses of remifentanil on the renal ischemiareperfusion (I/R) injury in rats. Methods Sixty male SD rats weighing 220-250 g were randomly divided into 5 groups ( n = 12 each): sham operation group (group S), model group (group M), low, median and high doses of remifentanil groups (RL, RM and RH groups). The rats were anesthetized with intraperitoneal 5% chloral hydrate 6 ml/kg. Renal ischemia was induced by clamping the bilateral renal arteries for 45 min using an atraumatwere infused via the caudal vein 15 min before ischemia respectively and the infusion was stopped at 30 min of reperfusion, while S and M groups received equal volume of normal saline instead. Blood samples were collected from the femoral vein at 30 min and 24 h of reperfusion for measurement of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations. The rats were sacrificed at 24 h of reperfusion and the renal tissues were removed for determination of MDA content, SOD and Ca2+ -ATPase activities. Pathological changes in renal tissues were observed with light and electron microscopes. Results Compared with group S, the concentrations of serum Cr and BUN and content of MDA were significantly increased, while activities of SOD and Ca2+ -ATPase were significantly decreased in the other 4 groups ( P ＜ 0.05 or 0.01). Compared with group M, the concentrations of serum Cr and BUN and content of MDA were significantly decreased, activities of SOD and Ca2+ -ATPase were significantly increased (P ＜0.05 or 0.01) and the pathological changes were reduced in RH, RM and RL groups. The plasma BUN and Cr concentrations and MDA content were decreased gradually and SOD and Ca2+ -ATPase activities were increased gradually with the increase in the doses of remifentanil in RL, RM and RH groups ( P ＜ 0.05 or 0.01 ).Remifentanil infusion significantly attenuated the pathologic changes in a dose-dependent manner. Conclusion Remifentanil can reduce the renal I/R injury in a dose-dependent manner by inhibiting lipid peroxidation and increasing Ca2+ -ATPase activity.     

骨髓间充质干细胞干预对缺血再灌注诱导肾小管上皮细胞凋亡的影响

目的：观察骨髓间充质干细胞（mesenchymal stemcells,MSCs）对缺血再灌注（ischemia/reperfusion,I/R）诱导的急性肾损伤模型小鼠肾小管上皮细胞（renal tubular epithelial cells,RTECs）凋亡的保护作用及机制。方法：Percoll密度梯度离心结合贴壁培养法有效分离出C57BL/6小鼠的骨髓间充质干细胞（mMSCs）。雄性C57BL/6小鼠45只,分为正常对照组（15只）、I/R组（15只、夹闭双侧肾蒂30min开放）、I/R＋mMSCs组（15只、夹闭双侧肾蒂30min开放的同时尾静脉注射mMSCs）。造模并干预后,于1d、2d、3d、7d、14d分别处死部分小鼠,留取动脉血及肾组织,检测血尿素氮（BUN）及肌酐（Scr）,制作肾组织切片行HE染色进行肾小管损伤程度评分,TUNEL法检测RTECs的凋亡,Western Blot法检测建模后第2天肾小管组织中Caspase-3、Bcl-2蛋白水平的表达。结果：I/R后小鼠的BUN及Scr均显著高于正常对照组,但同一时间点I/R＋mMSCs组小鼠的BUN及Scr水平较I/R组为低,以术后第7天差异最为显著（P〈0.01）,肾小管损伤程度评分也有着显著降低。TUNEL检测表明I/R可诱导RTECs发生明显的凋亡,以术后第2天凋亡指数（apoptosis index,AI）最高（46.71±11.20）（P〈0.01）,而mMSCs干预可显著降低各时间点RTECs的凋亡水平（P〈0.05或P〈0.01）,至14d时AI基本恢复至对照组水平。Western blot进一步显示：I/R＋mMSCs组小鼠的肾组织中Caspase-3蛋白水平明显下降（1.16±0.33,P〈0.01）,而Bcl-2水平却有明显增高（0.94±0.27,P〈0.01）。结论：移植的MSCs对I/R诱导的肾损害小鼠RTECs凋亡具有抑制作用,从而有利于肾小管损伤的早期恢复,其保护机制可能与抑制Caspase-3表达,上调Bcl-2表达有关,但其细胞学和分子机制还有待进一步研究。     

骨髓间充质干细胞干预对急性肾损伤鼠肾脏中细胞因子的影响

目的：建立缺血再灌注（I/R）诱导的急性肾损伤小鼠模型,探讨外源性地给予骨髓间充质干细胞（MSCs）干预对模型小鼠肾损伤微环境中细胞因子水平的影响,并探讨其在肾损害修复中的作用。方法：采用Percoll密度梯度离心法分离出C57BL/6小鼠的骨髓间充质干细胞（mMSCs）。雄性C57BL/6小鼠45只,分为正常对照组（15只）、I/R组（15只、夹闭双侧肾蒂30min开放）、I/R＋mMSCs组（15只、夹闭双侧肾蒂30min开放的同时尾静脉注射mMSCs）。于建模后1d、2d、3d、7d、14d分别处死部分小鼠（每次每组均处死3只）,留取动脉血及肾组织,检测血尿素氮（BUN）及肌酐（Scr）水平,制作肾组织切片行HE染色,观察肾组织病理变化,行肾小管坏死程度评分（ATN评分）。ELISA法检测肾组织匀浆肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、单核细胞趋化蛋白-1（MCP-1）、白细胞介素-10（IL-10）、肝细胞生长因子（HGF）、骨形态发生蛋白-7（BMP-7）的水平。结果：I/R组小鼠的BUN及Scr均显著高于正常对照组,肾小管损伤严重;而I/R＋mMSCs组小鼠的BUN及Scr水平较I/R组为低,以术后第7天差异最为显著（P〈0.01）,肾小管损伤病理明显减轻,ATN评分也有着显著降低。ELISA结果显示单纯I/R组各时间点肾组织匀浆中TNF-α、IL-1β、MCP-1的水平显著升高（P〈0.01或P〈0.05）,而IL-10、HGF、BMP-7的水平却有着显著降低（P〈0.01或P〈0.05）。但同时给予MSCs干预的I/R小鼠肾组织匀浆中前述细胞因子的水平却向着相反方向改变。结论：MSCs对I/R肾组织中细胞因子的分泌具有调控作用,而这些调控了的细胞因子进而可通过旁分泌作用发挥促进肾损害的修复作用。     

红细胞生成素对模拟急性肾损伤微环境下培养骨髓间充质干细胞增殖的影响及机制探讨

目的 观察经红细胞生成素（EPO）干预后,体外模拟急性肾损伤（AKI）微环境下培养骨髓间充质干细胞（MSC）的分裂增殖情况,并探讨产生这种变化的可能机制。 方法 抽取C57BL/6小鼠的骨髓,经Percoll密度梯度离心联合贴壁培养法分离纯化出小鼠MSC（mMSC）,以流式细胞仪鉴定。夹闭雄性C57BL/6小鼠双侧肾蒂30 min再开放30 min的方法制作AKI鼠模型,即刻取双侧肾脏皮质制作缺血再灌注（IR）肾脏匀浆上清。取扩增第3代的mMSC分组培养：A组：含10%胎牛血清的低糖DMEM培养基;B组：含10%胎牛血清的低糖DMEM培养基＋IR肾脏匀浆上清,体外模拟AKI微环境干预mMSC;C组：含10%胎牛血清的低糖DMEM培养基＋IR肾脏匀浆上清+不同浓度EPO（1、5、10、50 U/ml）。培养1、3、5、7 d。CCK-8法检测各组培养mMSC的增殖,TUNEL法检测mMSC凋亡,Western印迹法检测mMSC表面EPO受体（EPOR）及增殖凋亡相关信号通路蛋白的表达。 结果 CCK-8法检测显示,经IR肾脏匀浆上清干预后,不同时间点mMSC的增殖效应显著减弱（P ＜ 0.01）,而TUNEL检测表明其胞核染色阳性细胞的百分比显著高于A组（P ＜ 0.01）;给予EPO干预后,mMSC的增殖能力增强,胞核染色阳性细胞百分比降低,具有浓度依赖效应。Western印迹结果显示,第3代mMSC表面存在EPOR表达,培养5 d后EPOR相对表达量为0.092±0.015。EPO以剂量和时间依赖性降低AKI微环境下凋亡相关蛋白caspase-3的表达,同时上调凋亡抑制蛋白Bcl-2的表达。用10 U/ml EPO干预5 d后,相对于B组,磷酸化蛋白酪氨酸激酶2及磷酸化信号转导和转录因子的表达均显著上调（均P < 0.01）。 结论 EPO能促进体外AKI微环境干预下培养mMSC的增殖,减轻其凋亡,该效应经EPOR介导,并与增殖凋亡相关信号通路蛋白有关。     

大网膜包裹明胶-间充质干细胞复合物对马兜铃酸肾病保护作用的研究

目的研究大网膜包裹明胶-间充质干细胞(MSCs)复合物对马兜铃酸肾病(AAN)小鼠肾脏的修复和保护作用。 方法使用明胶与间充质干细胞共培养构建间充质干细胞-明胶复合物。用CCK-8法评价MSCs与明胶的生物相容性以及MSCs的增殖能力。将体重为20~25 g的健康雄性C57BL/6小鼠24只随机分为4组：正常对照组(Control组)、马兜铃酸肾病模型组(AAN组，腹腔注射马兜铃酸溶液)、大网膜包裹MSCs治疗组(MSCs组)、大网膜包裹MSCs-明胶复合物治疗组(MSCs-Gel组)。利用小动物活体成像系统观察MSCs在小鼠体内的分布和存活情况；检测各组小鼠的血肌酐、尿素氮水平变化，观察肾脏组织学变化及相关纤维化指标如α-SMA、TGF-β和胶原变化情况。 结果明胶对MSCs的增殖能力无明显影响。与单纯MSCs组相比，MSC-Gel组的MSCs在小鼠肾脏内存活时间更长，且血肌酐、尿素氮明显降低，肾组织病理改变纤维化程度明显减轻，α-SMA、TGF-β、Col-Ⅰ表达亦显著降低(均P<0.05)。 结论大网膜包裹明胶-MSCs复合物对AAN小鼠肾脏具有良好的修复和保护作用。     

骨髓间充质干细胞促进大鼠IgA肾病修复

目的 观察骨髓间充质干细胞（MSC）对大鼠IgA肾病有无修复作用，并探讨其可能的机制。 方法 SD大鼠随机分为MSC注射组、生理盐水（NS）组及健康对照组。前两组以牛血清白蛋白(BSA)+葡萄球菌肠毒素B（SEB）＋皮下注射四氯化碳（CCl4）的改良法建立IgA肾病模型。体外连续培养SD大鼠MSC并通过流式细胞仪和成骨成脂细胞诱导分化鉴定MSC，用5-溴脱氧尿嘧啶核苷（BrdU）体外标记培养的MSC。移植后1周及4周分别观察3组的体质量、尿蛋白量（24 h）、肾功能、肾脏病理变化、IgA荧光沉积变化；ELISA法检测尿中的MCP-1、TGF-β1量；RT-PCR法检测肾组织中MCP-1、TGF-β1 mRNA的表达情况；免疫组化观察细胞因子及BrdU标记的MSC在肾组织中的分布情况。 结果 移植后1周，MSC组尿蛋白量（24 h）（36.86±4.78） mg，Scr（53.50± 6.28） μmol/L；NS组尿蛋白量（24 h）(66.98±5.86) mg，Scr (82.50±8.36) μmol/L，两组差异有统计学意义（均P < 0.05）；同时，MSC组MCP-1、TGF-β1在尿中的含量及肾脏中表达均显著低于NS组（均P < 0.05）。移植后4周，MSC组体质量、肾脏病理变化、IgA荧光沉积与NS组差异有统计学意义；MCP-1、TGF-β1在尿中的含量及肾脏中的表达与健康对照组差异无统计学意义。随时间延长，BrdU标记的MSC在肾组织中分布却逐渐减少。 结论 MSC输注可促进大鼠IgA肾病的修复，其作用机制可能并不完全是依赖于MSC的直接分化，而是通过调节肾组织中细胞因子的分泌和（或）其他的功能进行修复。     

上调intermedin表达对大鼠肾脏缺血再灌注的保护作用

目的 观察上调肾脏intermedin( IMD)表达对大鼠肾脏缺血再灌注(I/R)的影响.方法 24只健康雄性Wistar大鼠随机分为假手术组、肾脏I/R组、IMD+I/R组、空质粒+I/R组,每组6只.所有动物于I/R术24h后杀检,取肾组织进行光镜检查,留取血清测定尿素氮(BUN)和血清肌酐(Scr)的浓度.免疫组织化学方法、半定量RT-PCR、Western印迹检测肾组织IMD表达及部位.Western印迹测定内皮素1(ET-1)、肿瘤坏死因子α(TNF-α)蛋白的表达.结果 HE、PAS染色结果显示,I/R组肾小管及间质病理损伤显著重于假手术组,IMD+I/R组小管间质损伤程度较肾脏I/R模型组及空质粒+I/R模型组明显减轻(1.5±0.8比7.6±2.3和7.0±1.8,均P＜0.05].与假手术组[(BUN 3.85±0.21 mmol/L,Scr(48.67±3.61) μmol/L相比,I/R组、IMD+I/R组以及空质粒+I/R组BUN(10.13±2.14) mmol/L,( 7.73±1.03) mmol/L,( 9.77±1.92) mmol/L和Scr(80.33±7.15) μmol/L,(58.50±:3.27)μmol/L,(75.67±7.58) μmol/L均明显升高(均P＜ 0.05),其中IMD+I/R组较I/R组以及空质粒+I/R组BUN和Scr水平显著降低(均P＜ 0.05).免疫组化结果显示,假手术组IMD呈弱阳性表达,主要位于肾小管间质细胞胞质内,I/R组肾组织IMD在肾小管上皮细胞和间质表达较假手术组上调;IMD+I/R组肾组织中IMD表达较I/R组明显上调(P＜0.01).与I/R组及空质粒+I/R组相比IMD+I/R组肾组织IMD mRNA,相对含量分别增加了60.7％、66.1％,蛋白相对含量分别增加了51.4％、55.9％.此外,与I/R组相比,IMD+I/R组肾组织ET-1、TNF-α蛋白表达显著降低(ET-1:0.17±0.02比0.08±0.02;TNF-α:0.35±0.02比0.21±0.04,均P＜0.05).结论 在大鼠肾脏I/R前上调IMD表达可能通过抑制ET-1、TNF-α表达保护肾脏结构及功能.     

骨髓间质干细胞通过影响PI3K/Akt信号通路抑制肝星状细胞的增殖

目的 探讨在体外非接触共培养条件下,大鼠骨髓间质干细胞(MSCs)如何调控肝星状细胞(HSCs)的增殖。方法 实验组将MSCs/HSCs按2×104/2×104(细胞/孔)的比例接种于Transwell共培养板上下层,建立非接触共培养体系;对照组为HSCs(2 ×104细胞/孔)单独培养;阳性对照组为加入抑制剂LY294002 (20 μmol/ml)观察抑制效果。共培养24、48、72h后应用流式细胞仪分析细胞周期,Western blot检测HSCs的p-Akt及Akt蛋白的表达。结果 共培养24、48、72 h后HSCs的S期细胞与对照组比较明显减少,且抑制程度呈时间依赖性(11.24±0.34 ＜15.73±0.76＜19.14±0.91 ＜23.16±1.80,P＜0.05);加入LY294002后可明显增加共培养组抑制效果,与单独使用抑制剂比较S期细胞减少更加明显(8.2±0.8＜11.7±1.6,P＜0.05);共培养组及共培养+抑制剂组p-Akt蛋白表达较单独培养组均显著下调,而Akt蛋白表达差异无统计学意义(P＞0.05);共培养+抑制剂组较共培养组p-Akt蛋白表达下调。结论 MSCs可明显抑制HSCs的增殖,可能是通过影响PI3K/Akt信号通路达到这一作用,磷酸化的Akt蛋白在其中起关键作用。     

mito-KATP通道在缺血后处理减轻大鼠肾缺血再灌注损伤中的作用

目的 评价线粒体ATP敏感性钾通道(mito-KATP通道)在缺血后处理减轻大鼠肾缺血再灌注损伤中的作用.方法 健康成年雄性SD大鼠35只,体重250～280 g,随机分为5组(n=7):假手术组(S组)仅分离双侧肾蒂,暴露45 min不夹闭;肾缺血再灌注组(I/R组)夹闭双侧肾蒂缺血45 min,再灌注6 h制备大鼠肾缺血再灌注模型;缺血后处理组(Ipo组)夹闭双侧肾蒂缺血45 min,再灌注10 s,缺血10 s,反复3次,再灌注6 h;mito-KATP通道阻断剂5-羟葵酸+I/R组(5-HD+I/R组)缺血前30 min腹腔注射5-HD 10 mg/kg,余处理同I/R组;缺血后处理+5-HD组(5-HD+Ipo组)缺血前30 min腹腔注射5-HD 10 mg/kg,余处理同Ipo组.于再灌注6 h时采集心脏血样,取肾并分离肾小管上皮细胞,测定血清Cr和BUN的浓度、肾小管上皮细胞线粒体膜电位、细胞内活性氧(ROS)含量和游离Ca2+浓度.结果 与S组比较,I/R组、Ipo组、5-HD+I/R组和5-HD+Ipo组血清Cr和BUN的浓度、肾小管上皮细胞内游离Ca2+浓度和ROS含量升高,线粒体膜电位降低(P＜0.05);与I/R组比较,Ipo组血清Cr和BUN的浓度、肾小管上皮细胞内游离Ca2+浓度和ROS含量降低,线粒体膜电位升高(P＜0.05),5-HD+I/R组和5-HD+Ipo组上述指标差异无统计学意义(P＞0.05);与Ipo组比较,5-HD+I/R组和5-HD+Ipo组血清Cr和BUN浓度、肾小管上皮细胞内游离Ca2+浓度和ROS含量升高,线粒体膜电位降低(P＜0.05).结论 mito-KATP通道的开放参与了缺血后处理减轻大鼠肾缺血再灌注损伤的过程.     

党参皂甙减轻大鼠移植肾缺血再灌注损伤中细胞凋亡的作用及其机制

目的 探讨党参提取物皂甙减轻移植肾缺血再灌注损伤中细胞凋亡的作用及其机制.方法 将雌、雄各半SD大鼠随机分成三组,每组20只,即假手术组、缺血再灌注组和皂甙干预组.假手术组不进行肾移植,仅切除右肾,游离左肾动静脉,暴露左肾1 h后关闭腹腔.缺血再灌注组和皂甙干预组建立大鼠移植肾缺血再灌注损伤模型.皂甙干预组分别于肾移植前48、24和0.5 h经腹腔注入皂甙溶液(每千克体重80 mg).移植肾再灌注后24 h,取大鼠外周血和移植肾组织待测.检测各组血尿素氮(BUN)和肌酐(Cr)水平;采用脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测各组肾组织原位细胞凋亡指数(AI);采用逆转录聚合酶链反应(RT-PCR)检测与细胞凋亡有关的基因Bcl-2和Bax mRNA在各组肾组织中的相对表达量.结果 与假手术组相比,缺血再灌注组和皂甙干预组血BUN和Cr水平都显著升高(P＜0.05);移植肾细胞凋亡指数也显著增高(P＜0.05);移植肾组织中Bcl-2 mRNA表达显著降低,Bax mRNA表达显著增高(P＜0.05).与缺血再灌注组比较,皂甙干预组血BUN和Cr值明显下降(P＜0.05);移植肾细胞凋亡指数明显下降(P＜0.05);移植肾组织中Bcl-2 mRNA表达显著增加,Bax mRNA表达明显下降(P＜0.05).结论 党参皂甙在移植肾缺血再灌注损伤中能显著减轻细胞凋亡.其机制可能是通过对Bcl-2基因表达的上调和对Bax基因表达的下调,从而抑制细胞的凋亡.