降钙素的镇痛作用及其对中脑导水管周围灰质中降钙素受体表达的影响

目的 探讨鲑鱼降钙素的镇痛作用及其对中脑导水管周围灰质(PAG)中降钙素受体表达的影响.方法 采用慢性压迫性损伤(CCI)的方法制备神经性疼痛的大鼠模型,热板实验评估热痛阈(TWL),von Frey纤丝测定机械痛阈(MNT),免疫组化法检测PAG中降钙素受体的分布,Western印迹检测降钙素受体蛋白的表达量.40只雄性SD大鼠被平均随机分配至5组:正常组、假手术组、CCI组、CCI+鲑鱼降钙素皮下注射组和CCI+鲑鱼降钙素PAG微量注射组.结果 (1)假手术组大鼠TWL、MNT及PAG中降钙素受体表达较正常组无显著性差异(P＞0.05);(2)CCI组大鼠TWL和MNT较正常组显著降低(P＜0.05),但降钙素受体表达显著高于正常组(P＜0.05);(3)鲑鱼降钙素治疗后CCI大鼠的TWL、MNT和降钙素受体的表达均显著升高(P＜0.05),且PAG注射较皮下注射效果显著(P＜0.05).结论 鲑鱼降钙素能够提高CCI大鼠的痛阈并增加PAG中降钙素受体的表达量,而且PAG注射鲑鱼降钙素较皮下注射效果明显.

Abstract：

Objective To investigate the analgesic effect of salmon calcitonin(sCT)and its effect on expression of calcitonin receptor(CT-R)in periaqueductal gray(PAG). Methods Rat models of neuropathic pain were prepared by chronic constriction injury(CCI). Thermal withdrawal latency(TWL)and mechanical nociceptive threshold(MNT)were measured using hot plate test and yon Frey monofilaments test. The distribution of CT-R in PAG was detected by immunohistochemical method. CT-R protein was quantitatively determined by western blotting. Fourty male SD rats were randomized into 5 groups: normal group, sham-CCI group, CCI group, CCI plussubcutaneous sCT group, and CCI plus microinjection of sCT into PAG group. Results TWL, MNT, andexpression of CT-R in PAG showed no difference between normal group and sham-CCI group(P＞0. 05). TWL and MNT in CCI group were significantly lower than those in normal group(P＜0.05), and expression of CT-R in CCI group was significantly higher than that in normal group(P＜0.05). TWL, MNT and expression of CT-R in CCI rats increased significantly after sCT therapy(P＜0. 05), and the effect was more marked in PAG injection group than subcutaneous injection group(P＜0.05). Conclusions sCT raises the pain threshold and increase the expression of CT-R in PAG of CCI rats, while PAG injection showed more marked effect than subcutaneous injection.     

追赶生长大鼠胃泌素及内脏脂肪细胞CCK2R表达变化

24只Wistar大鼠分为正常对照组、限食组、追赶生长组,检测所有大鼠血糖、血脂、血清胃泌素,内脏脂肪体脂比、脂肪细胞CCK2R mRNA和蛋白水平.结果 显示限食组和正常组相比,血清胃泌素水平降低54%(P＜0.05),内脏脂肪体脂比减少55%(P＜0.05),脂肪细胞CCK2R mRNA和蛋白表达下降(2.19±0.18对3.2±0.24,0.11±0.03对0.15±0.04,P＜0.05).追赶生长组血清胃泌素水平分别高于限食组72%和正常组31%(P＜0.05),内脏脂肪体脂比高于限食组114%(P＜0.05),达到正常对照组水平;同时脂肪细胞CCK2R mRNA和蛋白表达高于正常对照组(4.09±0.59对3.2±0.24,0.25±0.05对0.15±0.04,P＜0.05).追赶生长大鼠内脏脂肪优先沉积的机制可能和胃泌素分泌增加及脂肪细胞CCK2R表达增加相关.

Abstract：

Wistar rats(n=24) were divided into normal control group(NC), food restriction group(FR), and catch-up group(RN). Serum glucose,lipids, gastrin, the ratio of visceral fat to body fat, adipocyte CCK2R mRNA and protein levels were determined. Compared with NC group, FR rats had lower serum gastrin and visceral fat formation. The adipocyte CCK2R mRNA and protein levels of FR rats were lower than those of NC rats. Serum gastrin level of RN rats was higher than those of FR and NC rats(P＜0.05). The ratio of visceral fat to body fat in RN rats increased compared with FR rats and was close to that of NC rats. The adipocyte CCK2R mRNA and protein levels of RN rats were higher than those of FR and NC rats. Gastrin and its receptor pathway possibly play a role in the mechanism of visceral fat accumulation in catch-up rats.     

帕金森病运动并发症发生机制中G蛋白偶联受体激酶6与N-甲基-D-天冬氨酸受体关系的研究

目的 研究G蛋白偶联受体激酶6(GRK6)在帕金森病(PD)运动并发症发生机制中与N-甲基-D-天冬氨酸(NMDA)受体的关系.方法 建立PD运动并发症大鼠模型,25只大鼠分为3组.异动症(LID)组10只,腹腔注射左旋多巴甲酯23 d;MK-801处理组10只,第23天左旋多巴甲酯注射前腹腔注射MK-801;PD组5只,腹腔注射0.2%维生素C液.另设假手术组5只为对照组.观察MK-801处理左旋多巴诱导的运动并发症模型大鼠的行为变化,并用免疫组织化学方法和Western印迹方法检测大鼠纹状体区GRK6蛋白的表达情况.结果 PD组大鼠长期使用左旋多巴后出现明显的异常不自主运动,与人类LID具有相似特征.免疫组化结果显示.PD组损伤侧纹状体区GRK6阳性细胞指数减少至(4.81±1.31)×103(P＜0.05),LID组损伤侧GRK6阳性细胞指数进一步减少至(3.23±0.41)×103(P＜0.01).MK-801组LID大鼠异常不自主运动评分减少,药效期延长,同时损伤侧纹状体区GRK6阳性细胞指数增多至(4.64±1.39)×103(P＜0.05).Western印迹法检测结果同免疫组化基本相符,PD组损毁侧/未损毁侧纹状体区GRK6含量比值为(83.7±2.1)%,LID组GRK6值降低为(76.8±2.2)%,而MK-801组GRK6值升高至(91.1±2.7)%(P＜0.01).结论 NMDA受体拮抗剂能逆转大鼠运动并发症的发生,其机制可能与GRK6增多,抑制了谷氨酸受体的过度活化有关.

Abstract：

Objective To investigate the relationship between G protein-coupled receptor kinase 6 (GRK6) and N-methyl-D-aspartate (NMDA) receptor in the mechanism study underlying motor complications in Parkinson's disease (PD).Methods The rat models (n= 25) of Parkinsonism related motor complications were established and were randomly divided into levodopa-induced dyskinesia (LID) group (n= 10,intraperitoneal injection of levodopa for 23 d),MK-801 treatment group (n= 10,intraperitoneal injection of MK-801 at day23 after intraperitoneal injection with levodopa for 22 days) and PD group (n= 5,intraperitoneal injection of vitamin C).Another 5 rats were served as controls (sham-operation group).The behavior changes of rats in MK-801 treatment group were observed,and the expression of GRK6 in the striate of rats was detected by immunohistochemistry and Western blot.Results After the chronic treatment with levodopa methyl ester,PD rats displayed abnormal involuntary movements,which was similar to levodopainduced dyskinesia in PD patients.Immunohistochemistry showed that GRK6-positive cells of lesion side were decreased in LID rats as compared with PD rats [(3.23±0.41 ) × 103 vs.(4.81 ± 1.31 ) ×103,P＜0.01].Rats in MK-801 treatment group displayed the decreased AIM scores and increased peak rotation,and the increased GRK6-positive cells of lesion side as compared with LID rats (P＜0.05).Western blot showed that the levels of GRK6 was 83.7% ±2.1% in PD group (presented as lesion side/unlesion side),76.8% ± 2.2% in LID group and 91.1% ± 2.7% in MK-801 treatment group (intergroups comparison:all P＜0.05).These results were in accordance with the results of immunohistochemistry.Conclusions Antagonist of NMDA receptor can be used to reduce the motor complications in rats.It may be due to increased GRK6 which inhibits the overactivation of glutamic acid receptors.     

腺病毒过表达TXNIP对INS-1胰岛细胞损伤和凋亡的影响

目的 观察TXNIP过表达是否可以引起正常培养条件下的胰岛细胞发生损伤和凋亡,分析TXNIP引起细胞损伤和凋亡的下游途径,以进一步明确TXNIP在糖尿病中引起各器官细胞损伤的机制.方法 将使用正常糖脂浓度培养的胰岛细胞分为4组：正常培养组、Ad-eGFP空病毒组、Ad-TXNIP过表达组和Ad-TXNIPC247S点突变过表达组.结果 与Ad-eGFP空病毒组相比,两个过表达组的TXNIP mRNA水平、TXNIP蛋白水平均显著升高,表明病毒转染成功,TXNIP成功过表达.与空病毒组相比,Ad-TXNIP过表达组TXNIP与Trx的结合量升高（1.67±0.08比1.06±0.05,P＜0.05）,而Trx活性显著降低（0.42±0.11比1.13±0.14,P＜0.01）;LDH活性[（498.8±55.62）U/L比（266.2±36.49）U/L,P＜0.01]、caspase-3活性[（3.799±0.330）nmol·h-1·mg-1 pro比（0.979±0.100）nmol·h-1·mg-1 pro,P＜0.01]及p38激酶活性（2.45±0.22比1.10±0.08,P＜0.01）均显著增高.与Ad-TXNIP过表达组相比,C247S点突变的TXNIP过表达组TXNIP与Trx的结合量减少,Trx活性明显升高,LDH活性[（421.6±41.31）U/L比（498.8±55.62）U/L,P＜0.05]和caspase-3活性[（3.377±0.290）nmol·h-1·mg-1 pro比（3.799±0.330）nmol·h-1·mg-1 pro,P＜0.01]较低,p38激酶活性（0.80±0.07比2.45±0.22,P＞0.05）明显降低.结论 单纯过表达TXNIP可以引起正常糖脂浓度培养条件下的胰岛细胞发生损伤和凋亡.TXNIP介导细胞损伤和凋亡的机制一方面与其抑制Trx活性、增加自由基损伤、增加ASK1-p38激酶依赖的凋亡途径有关,同时也有不依赖结合抑制Trx的途径存在.TXNIP的表达上调是糖尿病引起胰岛细胞损伤和凋亡的重要原因.     

丁基苯酞对血管性痴呆大鼠记忆及海马Bcl-2、Bax的影响

目的 研究丁基苯酞(NBP)对血管性痴呆(VD)大鼠记忆、海马病理变化及抗凋亡蛋白(Bcl-2)和促凋亡蛋白(Bax)蛋白表达的影响.方法 采用永久性结扎双侧颈总动脉方法制备VD大鼠模型.SD大鼠被随机分成假手术组、VD模型组、NBP治疗组、尼莫地平治疗组.应用Morris水迷宫检测大鼠记忆能力,苏术精-伊红(HE)染色观察海马神经元形态,免疫组化检测海马Bcl-2和Bax的表达.结果 VD模型组与假手术组大鼠比较记忆能力显著下降,逃避潜伏期分别为(78.79±21.93) s与(16.96±7.44) s (P＜0.05),海马神经元病理改变严重,免疫阳性细胞数量显著增加,其中Bax变化更为显著 (43.00±6.72与6.00±1.29,P＜0.05),Bcl-2与Bax的比值较对照组明显降低;NBP治疗组与模型组比较记忆能力显著改善,逃避潜伏期分别为(47.13±21.75) s与(78.79±21.93) s (P＜0.05),海马神经元病理形态明显改善,Bcl-2免疫阳性细胞数显著增多(33.14±8.05与21.81±4.97,P＜0.05),Bax免疫阳性细胞数显著减少(32.93±4.99与43.00±6.72,P＜0.05).NBP治疗组与尼莫地平治疗组之间比较,各项指标均无显著差异 (P＞0.05).结论 NBP能改善VD大鼠记忆能力,抑制海马细胞凋亡,对VD大鼠有一定的治疗作用.

Abstract：

Objective To study the effects of butylphthalide (NBP) on memory and apoptosis related protein as well as neuronal pathology in hippocampus of vascular dementia (VD) rats. Methods VD model was generated by the permanent occlusion of bilateral common carotid arteries in SD rats to produce the forebran ischemia. Male SD rats were randomly allocated into sham-operation group, VD model group, NBP treatment group and nimodipine treatment group. The function of memory was tested by the Morris water maze. The neuronal pathological changes and the expression of Bcl-2 and Bax proteins in the hippocampus were observed with hematoxylin-eosin (HE) staining and immunohistochemical staining, respectively. Results The impaired memory of VD rats was proved by the lengthened mean escape latency [(78.79±21.93)vs.(16.96±7.44),P＜0.05] and the neuron in hippocampus was severely damaged. The decveased ratio of Bcl-2/Bax resulted from the overexpression of Bax proteins in VD model group versus the sham-operation group [(43.00±6.72)vs.(6.00±1.29),P＜0.05]. The treatment of NBP notably improved the memory function of VD rats and reduced the hippocampus pathological injury (P＜0.05). The expression of Bcl-2 protein raised [(33.14±8.05)vs.(21.81±4.97),P＜0.05] along with reduced expression of Bax protein [(32.93±4.99)vs.(43.00±6.72),P＜0.05] after NBP treatment. However, there was no significant difference in the treatment effects between nimodipine and NBP group (P＞0.05). Conclusions NBP treatment could improve memory of VD rats and reduce the hippocampus pathological lesion by inhibiting the apoptosis related protein.     

瑞舒伐他汀改善糖尿病大鼠血管内皮功能的实验研究

目的 评价不同剂量瑞舒伐他汀对糖尿病大鼠血管内皮功能的保护作用.方法 24只糖尿病大鼠随机分为3组:糖尿病对照组、瑞舒伐他汀20 mg组(RV 20 mg组)和10 mg组(RV 10mg组);8只健康SD大鼠作为正常对照组.给药共8周,分别于给药前和给药后第8周测定血浆内皮素-1(ET-1)、一氧化氮(NO)、血糖、血脂含量.结果 用药前糖尿病大鼠各组血糖均显著高于正常对照组(P＜0.01);RV 20 mg和RV 10 mg组在给药后第8周血糖稍低于糖尿病对照组,但差异无统计学意义(P＞0.05).糖尿病对照组血浆NO浓度明显低于SD正常对照组(P＜0.05),给药8周后,用药组血浆NO浓度明显高于糖尿病对照组(P＜0.01或P＜0.05).糖尿病对照组血浆ET-1浓度高于SD正常对照组(P＜0.05);给药8周后,用药组血浆ET-1浓度明显低于糖尿病对照组(P＜0.01或P＜0.05).RV 20 mg组和RV 10 mg组比较,血浆E-1浓度降低及血浆NO浓度升高(P＜0.05).同时,RV 20 mg组和RV 10 mg组血胆固醇水平明显低于糖尿病对照组(P＜0.01).结论 瑞舒伐他汀可明显降低糖尿病大鼠的血胆固醇水平,同时通过提高血浆NO浓度和降低E-1浓度等机制改善血管内皮功能.

Abstract：

Objective To evaluate the effects of two different dosage of rosuvastatin on endothelial dysfunction in diabetic rats. Methods The 24 diabetic rats were randomly divided into three groups (n=8,each): diabetic control group, 20 mg rosuvastatin daily (RV 20 mg group) and 10mg rosuvastatin daily for 8 weeks (RV 10 mg group) and normal control group (SD group). The levels of blood glucose, lipid, nitric oxide(NO) and endothelin-1 (ET-1) were measured before and 8 weeks after treatment. Results The levels of blood glucose were higher in all diabetic rats groups than in SD group before experiment (P＜0. 01). Compared with diabetic rats control group, blood glucose was slightly lower in RV 10 mg group and RV 20 mg group at 8 weeks (P＞0. 05). The plasma NO level was significantly lower in diabetic rats control group than in SD group (P＜0. 05).After 8 weeks, plasma NO levels were significantly higher in RV 20 mg and RV 10 mg groups than in diabetic rats control group (P＜0. 01 or P＜0. 05). The plasma levels of ET-1 was significantly higher in diabetic rats control group than in SD group (P＜0. 01). After 8 weeks, plasma ET-1 levels were significantly lower in RV 20 mg and RV 10 mg group than in diabetic rats control group (P＜0. 01).Meanwhile, the plasma lipids were lower in RV 20 mg and RV 10 mg group than in diabetic control group (P＜0. 05 or P＜0. 01). Conclusions Rosuvastatin can adjust blood lipids and significantly improve endothelial function in diabetic rats by increasing plasma NO level and decreasing plasma ET-1 level.     

N-乙酰半胱氨酸对糖尿病肾病大鼠肾皮质转化生长因子表达的影响

目的 探讨N-乙酰半胱氨酸(NAC)对糖尿病肾病(DN)大鼠肾皮质转化生长因子(TGF-β1)表达的影响.方法 制备DN大鼠模型,将动物随机分为正常对照组、DN组和NAC组.8周后测定大鼠尿白蛋白排泄率,免疫组化和RT-PCR方法检测肾皮质TGF-β1的表达水平,观察肾脏标本病理变化.结果 (1)DN组和NAC组尿白蛋白排泄率与正常对照组比较明显增多[(1268.3±297.5)μg/24 h和(315.9±86.8)μg/24 h比(31.2±8.9)μg/24 h,q值分别为29.85、16.76,均P＜0.01];肾皮质TGF-β1表达水平升高,肾小球系膜区免疫组化指数7.35±1.17和3.87±0.71比1.95±0.34(q值分别为l0.75、5.82,均P＜0.01),肾小管间质区免疫组化指数21.21±3.78和10.67±1.86比3.62±0.79(q值分别为15.20、11.36,均P＜0.01),肾皮质mRNA表达量0.72±0.06和0.45±0.05比0.23±0.04(q值分别为9.13、7.45,均P＜0.01),肾脏病理改变明显;(2)与DN组比较,NAC组尿白蛋白排泄率减少(q=8.17,P＜0.01),肾皮质TGF-β1的表达水平降低[肾小球系膜区免疫组化指数(q=4.97,P＜0.01),肾小管间质区免疫组化指数(q=6.86,P＜0.01),肾皮质mRNA表达量(q=3.69,P＜0.05)]和肾脏病理病变轻;(3)NAC组大鼠肾皮质TGF-β1 mRNA的表达量与尿白蛋白排泄率水平正相关(r=0.749,P＜0.05).结论 NAC对DN大鼠肾脏有保护作用,其部分机制与抑制TGF-β1表达有关.

Abstract：

Objective To investigate the effects of N-acetylcysteine (NAC) on the expression of transforming growth factor-β1 (TGF-β1) in renal cortex of diabetic nephropathy rats.Methods A rat model of DN was established.The rats were randomly divided into control group,DN group and NAC group.After 8 weeks treatment,urinary albumin excretion rate (UAER) was determined.The expression of TGF-β1 in renal cortex was detected by immunohistochemistry and RT-PCR analysis.Pathomorphological changes of renal cortex were observed.Results (1)The levels of UA ER were significantly higher in DN group and NAC group [(1268.3±297.5) μg/24 h and (315.9-±86.8) μg/24 h] than in control group [(31.2±8.9) μg/24 h,q-29.85,16.76,both P＜0.01].The groups of DN and NAC versus group of control showed the increased levels of activity of TGF-β1 in renal cortex [immune-histochemistry index of glomerular mesangial area:7.35±1.17 and 3.87 ± 0.71 vs.1.95±0.34,q= 10.75,5.82,both P＜0.01];immune-histochemistry index of renal tubulointerstitium [21.21± 3.78 and 10.67±1.86 vs.3.62±0.79,q=15.20,11.36,both P＜0.01];the expression of mRNA in renal cortex[0.72±0.06 and 0.45±0.05 vs.0.23±0.04,q=9.13,7.45,both P＜0.01].The pathomorphological changes were significant in DN group and NAC group.(2)The NAC group versus DN group showed a decreased levels of UAER (q=8.17,P＜0.01),activity of TGF-β1 in renal cortex [immune-histochemistry index of glomerular mesangial area:q= 4.97,P＜0.01]immune-histochemistry index of renal tubulointerstitium (q = 6.86,P ＜ 0.01 );the expression of mRNA in renal cortex (q= 3.69,P＜0.05) and showed improvement of pathomorphology in renal cortex.(3) There was a significantly positive correlation between expression quantity of TGF-β1 mRNA in renal cortex and UAER level in NAC group(r= 0.749,P＜0.05).Conclusions The protective effects of NAC on the kidney of DN rats may be partly related with inhibition on the expression of TGF-β1.     

Protective effect of sodium tanshinon Ⅱ A on lung injury induced by limb ischemia-reperfusion in rats

Objective To investigate the protective effect of sodium tanshinone Ⅱ A on lung injury induced by limb ischemia-reperfusion in rats. Methods 60 SD rats were randomly divided into three groups:control group (group A,n =20),limb-ischemia-reperfusion group (group B,n =20),and tanshinone Ⅱ A group (group C,n =20). The levels of malondialdehyde (MDA),superoxide dismutase (SOD),xanthine oxidase (XOD),interleukin-8 (IL-8),and tumor necrosis factor-α (TNF-α) in plasma and lung tissue were measured. The levels of TXB2 and 6-kcto-PGF1α in serum were measured by radioimmunoassay. The morphological and ultrastructure changes of lung tissues were observed under light microscope and electron microscope,respectively. Results ①Under light microscope,compared with group B,expansion of pulmonary vascular,aggregation of inflammatory cell,pulmonary edema relieved in group C. Under electron microscopy,compared with group B,mitochondrial swelling or degeneration,edema of basement membrane,swelling of pulmonary capillary endothelial cell,and pulmonary interstitial edema relieved in group C. ②MDA and XOD in plasma and lung tissue of group B were significantly higher than those in group A (all P ＜0. 01),and those in group C were significantly lower than those in group B (all P ＜0. 01). SOD in plasma and lung tissue of group B was significantly lower than that in group A (all P ＜0. 01). and that in group C was significantly higher than that in group B (all P ＜0. 01). ③IL-8、TNF-α in serum and lung tissue of group B were significantly higher than those in group A (all P ＜0. 01),those in group C were lower than those in group B ( P ＜0. 05 or P＜0. 01),and there was no statistical significance between group C and group A. ④TXB2 and 6-keto-PGF1α in serum of group B were significantly higher than those in group A (all P ＜0. 01),and those in group C were significantly lower than those in group B (all P ＜0. 01). TXB2 in serum of group C was higher than that in group A ( P ＜0. 05). There was no statistical significance on serum 6-keto-PGF1α between group C and group A. Conclusions Tanshinone Ⅱ A can alleviate lung injury of rats with limb ischemia-reperfusion.     

还原型谷胱甘肽对糖尿病大鼠肾脏保护作用的机制研究

目的 探讨还原型谷胱甘肽(reduced glutathione,GSH)对链脲佐菌素(streptozotocin,STZ)诱导的糖尿病大鼠肾脏线粒体保护及机制.方法 STZ诱导糖尿病大鼠模型,将糖尿病大鼠随机分为糖尿病非干预组(DM组)和GSH干预组(DM+GSH组),并以正常组(NC组)作对照.干预8周后,测定各组大鼠尿白蛋白排泄率(UAER)、血肌酐(SCr)、血尿素氮(BUN)水平,光镜观察肾脏组织形态学变化,检测血清和肾皮质中丙二醛(MDA)、超氧化物歧化酶(SOD)的含量以及各组肾脏线粒体膜电位和肿胀度的变化.结果 DM组大鼠UAER、SCr、BUN较NC组显著升高(均P＜0.05),肾脏组织发生糖尿病肾病病理改变,血清MDA(μmol/L)和肾皮质中MDA(μmol/g)显著升高(5.59±1.03 vs 2.97±0.77;4.80±0.83 vs 2.98±0.75;均P＜0.01),血清SOD(U/ml)和肾皮质中SOD(U/mg)含量显著降低(89.13±22.73 vs 124.1±9.27;46.05±10.24 vs 89.89±17.62;均P＜0.01),肾脏线粒体膜电位明显降低(495.79±124.71 vs 965.77±246.48,P＜0.05),线粒体肿胀度趋势明显减弱.与DM组相比,DM+GSH组大鼠UAER、SCr、BUN显著降低(均P＜0.05),肾脏病理形态得到一定改善.血清MDA(μmol/L)和肾皮质中MDA(μmol/g)降低(4.15±0.59 vs 5.59±1.03;3.39±0.61 vs 4.80±0.83;均P＜0.05),血清SOD(U/ml)及肾皮质中SOD(U/mg)升高(112.92±8.93 vs 89.13±22.73;83.15±16.75 vs 46.05±10.24;均P＜0.05),肾脏线粒体膜电位显著升高(715.97±188.65 vs 495.79±124.71,P＜0.05),线粒体肿胀度趋势增强.结论 还原型谷胱甘肽对STZ诱导的糖尿病大鼠肾脏病变有一定程度的保护作用,其机制可能与线粒体的功能改变有一定关系.     

糖尿病周围神经病变患者足底压力变化

目的 了解2型糖尿病合并周围神经病变患者足底压力变化特点.方法 根据患者有无合并周围神经病变将2004年1月至2009年12月诊治的1103例2型糖尿病患者分为合并周围神经病变组（DPN组,n=301）和无周围神经病变组（DC组,n=802）.记录患者一般资料;测定血脂谱、空腹血糖和糖化血红蛋白水平及尿蛋白排泄率;用足底压力测量仪一步法测量两脚各5次动态足底压力,计算足底压力参数值.对研究数据采用独立样本t检验或Mann-Whitney U检验进行统计分析.结果 相比DC组,DPN组患者年龄大、腰臀围比值大、收缩压高、空腹血糖和糖化血红蛋白水平高、尿白蛋白排泄率高（均P＜0.05）.两组足底峰值压力差异无统计学意义（P＞0.05）;但DPN组接触时间延长[分别为（1484±412）和（1241±281）ms,t=-9.414,P＜0.05],压力-时间积分[分别为（333±115）和（278±89）kPa·s,t=-7.446,P＜0.05]和应力-时间积分[分别为（628±187）和（536±149）N·s,t=-7.707,P＜0.05]增加.与DC组相比,DPN组足跟[分别为（396±101）和（411±105）kPa,t=2.163,P＜0.05]和第2跖骨头[分别为（240±87）和（269±95）kPa,t=4.563,P＜0.05]、第3跖骨头[分别为（241±75）和（262±77）kPa,t=4.046,P＜0.05]峰值压力降低,但足弓[分别为（122±48）和（115±31）kPa,t=-2.487,P＜0.05]和第5跖骨头[分别为（218±116）和（195±99）kPa,t=-3.131,P＜0.05]及第3～5趾区域[分别为（108±50）和（98±46）kPa,t=-3.315,P＜0.01]峰值压力增高.而与DC组相比,DPN组足跟[分别为（228±100）和（189±67）kPa·s,t=-6.201,P＜0.05]、足弓[分别为（82±45）和（66±26）kPa·s,t=-6.151,P＜0.05]及前足底各个区域的压力-时间积分均明显增高.结论糖尿病周围神经病变患者足底压力分布异常、承受压力时间延长;两种因素共同作用致足底压力-时间积分增高,后者可致糖尿病足压力性溃疡.     

2型糖尿病慢性肾脏疾病患者血浆心钠素水平的变化及其临床意义

目的 探讨不同时期糖尿病慢性肾脏疾病（CKD）患者血浆心钠素（ANP）水平变化及其临床意义. 方法 选取T2DM患者93例,根据UAER分为单纯糖尿病（DM）组30例、CKD 3期（CKD3）组31例及CKD 4期（CKD 4）组32例,另选健康体检者30名作为正常对照（NC）组.采用ELISA测定各组血浆ANP水平. 结果 （1）DM、CKD 3和CKD 4组ANP水平均高于NC组,且随UAER增加而增加[（280.89±43.66） vs （356.87±45.45） vs （414.95±57.52） vs （164.09±36.64） ng/L,P＜0.05];（2）ANP水平与病程、HbA1c、FPG、TG、BUN、Cr呈正相关（r=0.77、0.82、0.73、0.71、0.80、0.74,P均＜0.05）,与HDL-C呈负相关（r=-0.68,P＜0.05）. 结论 血浆ANP与CKD患者UAER正相关,FPG、HbA1c、BUN、Cr和TG是血浆ANP的独立正相关因子.     

普罗布考对糖尿病大鼠大血管病变的影响

目的 探讨普罗布考对糖尿病大鼠大血管病变的影响及机制.方法 44只健康Sprague Dawley(SD)雄性大鼠随机分为对照组(NC组,n=8)、糖尿病组(DM组,n=18)、普罗布考干预组(PB组,n=18)3组,四氧嘧啶50 mg/kg行尾静脉注射制备糖尿病大鼠模型.糖尿病造模成功后PB组给予普罗布考170 mg...     

2型糖尿病大鼠肺组织内皮素-1、降钙素基因相关肽及P物质水平变化及其对肺动脉高压的影响

目的 探讨2型糖尿病大鼠肺组织内皮素-1(ET-1)、降钙素基因相关肽(CGRP)、P物质(SP)的变化.方法 3个月龄健康雄性Wistar大鼠10只,采用高脂饲料喂养联合30μg/g链脲佐菌素尾静脉注射制备2型糖尿病大鼠模型.另设正常对照组(n=8),给予普通饮食.造模成功后12周末股动脉放血处死大鼠,取右下肺组织行Weigert来复红弹力纤维染色,计算与呼吸性细支气管、肺泡管伴行的肺小动脉管壁厚度占血管外径百分比(WT%)以及管壁面积占血管总面积百分比(WA%).应用免疫组织化学方法观察肺组织ET-1、CGRP、SP表达水平.数据统计采用独立样本t检验.结果 与正常对照组比较,糖尿病组大鼠肺泡隔面积与视场总面积比(0.42±0.10 vs 0.29±0.06,t=5.842,P＜0.05)、肺动脉WT%(26%±8%vs 19%±9%,t=3.023,P＜0.05)和WA%(42%±8% vs 36%±9%,t=2.526,P＜0.05)均显著增加,ET-1、SP平均吸光度(ET-1:86±5 vs 83±4,t=2.402,P＜0.05;SP:101±4 vs 100±3,t=2.530,P＜0.05)和阳性面积/视场总面积比(ET-1:0.016±0.017 vs0.010±0.008,t=2.501,P＜0.05;SP:0.014±0.014 vs 0.009±0.009,t=2.127,P＜0.05)显著增加,CGRP平均吸光度和阳性面积/视场总面积比差异无统计学意义.结论 糖尿病大鼠存在肺动脉高压的病理结构改变,可能与其肺组织ET-1表达增加有关.     

Chronic intermittent hypoxia induces thioredoxin system changes in a gender-specific fashion in mice

Obstructive sleep apnea (OSA) is an independent risk factor of multisystem injury including liver and cardiovascular system. Chronic intermittent hypoxia (CIH) associated with recurrent apneas in patients with OSA is one of the most important causes of the increased various systems injury and oxidative stress induced by CIH is an important pathogenic mechanism. Reports indicated that females are less susceptible to oxidative stress injury. The goal of this study was to explore if there exists gender deference of thioredoxin system (Trx/Txnip) alterations by CIH and to clarify a clue for studying gender disparity of OSA-related multisystem injury. C57BL/6J mice of each gender were exposed to CIH with a fractional inspired O2 (FiO2) nadir of 5%. The oxidative and antioxidant biomarkers were evaluated, including serum OxLDL level and Trx/Txnip expression of liver tissue. Male mice exposed to CIH exhibited significant increases in serum OxLDL level than that of the male control (73.24 ± 22.43 μg/dL, 45.20 ± 28.53 μg/dL, P = 0.032) but no significant difference in the females. Male mice exposed to CIH also exhibited decreased expression of Trx than the female (0.4460 ± 0.1023 versus 1.0454 ± 0.1777, P = 0.013) and increased expression of Txnip than the female (0.0123 ± 0.0476 versus 0.0065 ± 0.0058, P = 0.022). These data suggest that CIH induces thioredoxin system change in a gender-specific fashion in mice.     

普罗布考与普伐他汀预防经皮冠状动脉介入治疗后再狭窄的对比研究

目的评价普罗布考预防经皮冠状动脉介入治疗（PCI）后再狭窄的作用。方法将准备行PCI的82例稳定型心绞痛患者随机分为普罗布考组（n=42）和普伐他汀组（n=40）。两组患者于术前4周开始服用普罗布考1 000 mg/d或普伐他汀40 mg/d。服药4周后行金属裸支架置入术。术后继续原剂量及方法服用药物至24周。出院后定期随访,术后24周复查冠状动脉造影。结果PCI后随访至24周,两组严重临床事件发生率（死亡、急性心肌梗死、卒中及紧急血运重建术）差异无统计学意义（P〉0.05）。复查冠状动脉造影结果显示普罗布考组再狭窄率（22.5%）低于普伐他汀组（36.8%,P〈0.05）,普罗布考组管腔直径狭窄率及晚期管腔丢失指数（分别为23.25%±10.08%及0.25±0.41）均低于普伐他汀组（分别为34.76%±16.99%及0.42±0.68,P〈0.05）,纯获得（2.11±1.02 mm）大于普伐他汀组（1.51±0.96 mm,P〈0.05）。而两组晚期管腔丢失比较则差异无统计学意义（P〉0.05）。结论PCI前4周应用普罗布考降低PCI后再狭窄率的作用优于普伐他汀。     

乙肝后肝硬化合并肝性脑病的影响因素

目的:探讨影响乙肝后肝硬化合并肝性脑病(hepatic encephalopathy,HE)发生、发展的相关因素,为预防HE发生及改善预后提供依据.方法:回顾性分析2002-03/2011-05在大连医科大学附属第一医院收治的乙肝后肝硬化合并HE患者共87例.同时选取同期住院的乙肝后肝硬化未合并HE患者52例作为对照.分别统计入院时的临床表现,根据WestHaven标准和Child-Pugh分级进行HE分期和肝功能分级,记录入院后各生化指标的第一次结果,包括血氨、血钠、血清胆碱酯酶、血清总胆红素、血清白蛋白、血尿素氮、血清肌酐、凝血酶原时间.对以上资料进行对比分析研究,寻找影响乙肝后肝硬化HE发生、发展的相关因素.结果:因HE入院患者中,感染位居HE诱因首位(35.6%);HEⅡ期所占比例最高(36.8%),Ⅳ期居第2位(25.3%).合并HE组与未合并HE组比较肝功能分级、腹水、血氨、血钠、血清胆碱酯酶、血清总胆红素、血清白蛋白、血尿素氮、凝血酶原时间均有统计学差异(0:8:79vs0:21:31,19:68vs36:16,20.74±70.44vs46.53±10.67,136.01±6.65vs141.48±3.34,1927.34±70.52vs3342.01±38.03,91.75±84.56vs58.98±14.46,25.53±3.84vs29.48±2.58,13.57±14.76vs6.41±1.57,21.34±7.12vs18.59±1.66;bothP<0.05).肝功能程度与HE分期无关,合并HE患者血氨水平在Ⅳ期较其他各期显著升高(156.91±62.94vs104.13±73.07,112.69±60.87,104.67±82.00;bothP<0.05).血钠水平在Ⅱ期、Ⅲ期、Ⅳ期较Ⅰ期显著降低(135.22±6.05,134.91±7.79,134.55±6.25vs139.73±5.15;bothP<0.05),血清胆碱酯酶水平在Ⅳ期较Ⅰ期显著降低(1605.19±76.01vs2325.46±71.31,P<0.05).血尿素氮水平在Ⅳ期较Ⅰ期、Ⅱ期显著升高(21.65±23.69vs6.53±3.41,10.62±7.37;bothP<0.05).结论:在因HE入院患者中,感染居乙肝后肝硬化发生HE诱因的首位.肝功能C级、合并腹水、高血氨、高血清总胆红素、高血尿素氮、低血钠、低血清胆碱酯酶、低血清白蛋白和凝血酶原时间延长可增加HE的发生率.高血氨、高血尿素氮、低血钠和低血清胆碱酯酶可加重HE的程度,并影响预后.     

雷公藤甲素对2型糖尿病大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及机制

目的 观察雷公藤甲素对2型糖尿病(T2DM)大鼠肾组织足细胞Nephrin、Podocin蛋白表达的影响及其机制探讨.方法 SPF级雄性8周龄Wistar大鼠50只,体重(200±20)g,按随机数字表法分为2组:对照组10只、模型组40只.模型组高脂高糖喂养8周联合小剂量链脲佐菌素(STZ)30mg/kg建立T2DM模型鼠,正常组大鼠喂养常规饲料.将造模成功的T2DM大鼠(28只)随机分成2组:糖尿病组(14只)和雷公藤甲素治疗组(14只),雷公藤甲素治疗8周后,检测大鼠生化指标、肾重/体重和尿白蛋白.利用光镜、电镜观察肾脏病理改变.采用实时定量PCR、免疫组化及Western blot法检测肾组织Nephrin及Podocin蛋白、骨桥蛋白、转化生长因子-β1(TGF-β1)及单核/巨噬细胞表面特异性标志抗原(ED-1)的mRNA和蛋白表达分布.采用单因素方差分析法进行统计学分析.结果 糖尿病大鼠尿白蛋白明显高于对照组(F=181.51,P＜0.01),治疗组尿白蛋白明显低于DM组(F=181.51,P＜0.01).与对照组比较,糖尿病大鼠肾组织Nephrin(F=36.82,P＜0.05)和Podocin(F=32.57,P＜0.05)蛋白表达下调,Nephrin(F=88.45,P＜0.01)、Podocin(F=55.43,P＜0.01)mRNA表达下调;雷公藤甲素组干预8周后Nephrin[(1.82±0.06)和(1.63±0.06),P＜0.05]和Podocin[(1.80±0.06)和(1.60±0.06),P＜0.05]蛋白质表达上调,肾脏组织Nephrin[(0.73±0.12)和(0.19±0.08),P＜0.01]和Podocin[(0.60±0.12)和(0.26±0.07),P＜0.01]mRNA表达上调.与对照组比较,T2DM大鼠肾组织骨桥蛋白(F=40.06,P＜0.05)和TGF-β1的(F=28.84,P＜0.05)蛋白质表达上调,肾组织骨桥蛋白(F=177.46,P＜0.01)和TGF-β1(F=161.27,P＜0.01)mRNA表达上调,雷公藤甲素治疗后肾组织骨桥蛋白[(1.27±0.03)和(1.39±0.05),P＜0.05]和TGF-β1[(1.23±0.03)和(1.44±0.02),P＜0.05]的蛋白表达下调,肾组织骨桥蛋白[(2.05±0.16)和(3.26±0.26),P＜0.01]和TGF-β1[(4.0±0.70)和(6.5±0.71),P＜0.01]mRNA表达下调.结论 雷公藤甲素对T2DM大鼠足细胞有保护作用,其机制可能与其?     

链霉蛋白酶对胃组织中治疗幽门螺杆菌药物浓度的影响

目的 明确链霉蛋白酶应用对胃组织中阿莫西林和甲硝唑浓度的影响.方法 C57BL/6小鼠随机分为实验组和对照组(每组70只),给予奥美拉唑(138.2 mg/kg)+阿莫西林(28.6 mg/kg)+甲硝唑(22.5 mg/kg),并分别予链霉蛋白酶(110 mg/kg)或等量的无菌PBS,上述药物灌胃1次,给药后15～360 min共10个时间点(每组7只/时间点)取血浆和胃组织,高效液相色谱法检测阿莫西林和甲硝唑浓度,并检测给药后120、360min的胃组织黏膜层胃炎指数(HE法)和黏蛋白5AC的相对表达量(蛋白质印迹法).结果 阿莫西林和甲硝唑的胃组织浓度达峰时间早于在血浆出现的时间(均为15 min比60 min),实验组的胃组织浓度明显高于对照组(P＜0.05),主要集中在15～90 min.实验组药物血浆浓度在15和30 min较对照组高(P＜0.05).给药后120、360 min,实验组与对照组的胃黏膜层胃炎指数的差异均无统计学意义(0.28±0.18比0.14±0.14,P＞0.05;0.43 ±0.20比0.28 ±0.18,P＞0.05);黏蛋白5AC的相对表达量实验组较对照组低(0.036 ±0.006比0.197±0.058,P＜0.05;0.039±0.008比0.208±0.072,P＜0.05).结论 链霉蛋白酶明显增加了抗菌药物从胃腔向胃黏膜方向的弥散量,使局部胃组织和血浆浓度升高,且以局部胃组织浓度升高为主,持续时间更长.

Abstract：

Objective To evaluate the effect of pronase on amoxicillin and metronidazole concentrations in gastric tissue. Methods C57BL/6 mice were randomly divided into experimental group ( n = 70 ) and control group ( n = 70 ) . Amoxicillin ( 28. 6 mg/kg ) , metronidazole ( 22. 5 mg/kg) and omeprazole (138.2 mg/kg) were administered orally to C57BL/6 mice, combined with pronase (110 mg/kg) or same amount of sterile PBS. Gastric tissue and blood plasma samples were taken at 10 point-in time (7 mice/time) from 15 min up to 360 min after administration. Concentrations of amoxicillin and metronidazole were detected by high performance liquid chromatography. Gastritis index of gastric mucosa ( hematoxylin-eosin staining) and the gastric tissue expressions of mucin 5 AC (Western blot) were detected at 120 min and 360 min after administration. Results The time to peak concentration of amoxicillin and metronidazole in gastric tissue appeared earlier than that in blood plasma (15 min vs 60 min). Tissue concentrations of amoxicillin and metronidazole of experimental group were significantly higher than those of control, and they were mainly at 15 min to 90 min (P ＜0. 05). Plasma concentrations of amoxicillin and metronidazole of experimental group at 15 min and 30 min were higher than those of control ( P ＜ 0. 05 ). There was no difference in gastritis index between experimental group and control at 120 min and 360 min after administration (0.28±0. 18 vs 0. 14 ±0. 14,P＞0.05; 0. 43 ±0. 20 vs 0. 28 ±0. 18,P ＞0. 05). The expressions of mucin 5 AC in experimental group were lower than those of control ( 0. 036 ± 0. 006 vs 0. 197 ± 0. 058; P ＜0. 05; 0. 039 ± 0. 008 vs 0. 208 ± 0. 072, P ＜ 0. 05 ). Conclusions Pronase can significantly enhance the drugs penetration from mucus into gastric tissue. Concentrations of amoxicillin and metronidazole of experimental group in local gastric tissue and plasma are higher than those of control, especially in improving concentrations of gastric tissue and prolongation of exposed time.