Effect of suiphated glycosaminoglycans on albuminuria in patients with overt diabetic (type 1) nephropathy

Decreased expression of heparan sulphate has been shown in theglomerular basement membrane of patients with overt diabeticnephropathy. Low-molecular-weight heparin (LMWH) is a highlysulphated glycosaminoglycan with strong structural and functionalsimilarities to heparan sulphate. In a first study, we set outto assess if LMWH could decrease the urinary albumin excretionrate (AER) in diabetic patients with overt nephropathy. Sixpatients entered a randomized, double-blind, placebo-controlledcrossover study with treatment episodes of 1 month, separatedby a 1-month wash-out. Patients self-administered prefilledsyringes with either placebo or LMWH (enoxaparin 40 mg/0.4 ml)at bedtime. Baseline AER levels before either treatment periodwere similar. In contrast to placebo, AER significantly decreasedfrom 447 (181—1102) to 295 (100—873) µg/minafter 1 month treatment with LMWH (P<0.05). Compared to placebo,the effect of LMWH did not reach statistical significance inthese six patients after 1 month treatment (P=0.16). Haemodynamicvariables including glomerular filtration rate and filtrationfraction did not change during enoxaparin treatment. We observed a favourable effect on AER during LMWH treatmentin diabetic patients with overt nephropathy. These data suggestthat long-term treatment trials in a larger group of patientsmay potentially demonstrate a new therapeutic option for patientswith overt diabetic nephropathy.     

Risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis and diabetic nephropathy: A 3-year follow-up study

This study aimed to explore the risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis (DFO) and diabetic nephropathy (DN). This is a prospective cohort study. Between May 2018 and May 2021, we selected 120 inpatients with comorbid severe diabetic foot infection (PEDIS Grade 3 or above) and DN for inclusion in our study. All cases were followed up for 36 months. The study outcomes were whether foot ulcer recurred and the time to recurrence. The risk factors of ulcer recurrence were analysed by comparing the data of the three groups. According to the recurrence of foot ulcer, the participants were divided into three groups: Group A (no foot ulcer recurrence, n = 89), Group B (foot ulcer recurrence within 12-36 months, n = 19) and Group C (foot ulcer recurrence within 6-12 months, n = 12). The multivariate Cox regression analysis showed that urine albumin-creatinine ratio (UACR) (HR: 1.008, 95% CI: 1.005-1.011, P < .001) and vibration perception threshold (VPT) (HR: 1.064, 95% CI: 1.032-1.096, P < .001) were identified as independent risk factors. Kaplan-Meier curves showed a significant positive association between UACR or VPT and the risk of foot ulcer recurrence (log rank, all P < .05). Areas under the ROC curves for UACR, VPT and the combination of UACR and VPT were 0.802, 0.799 and 0.842, respectively. The best cut-off values of UACR and VPT were 281.51 mg/g and 25.12 V, respectively. In summary, elevated UACR and VPT were independent risk factors. The best clinical cut-off values of UACR and VPT for prediction of foot ulcer recurrence were 281.51 mg/g and 25.12 V, respectively. Besides, our results suggested that microcirculation disorders rather than macrovascular complications play a major role in the recurrence of foot ulcer in patients with comorbid DFO and DN.     

Effect of prednisolone therapy on serum concentrations of soluble interleukin-2 receptor in patients with IgA nephropathy

Background It is not known whether steroid therapy affects the serum level of soluble interleukin-2 receptor (sIL-2R) in patients with glomerulonephritis. We therefore examined the relationship between serum level of sIL-2R and indicators of disease activity in patients with immunoglobulin (Ig) A nephropathy, and serum levels of sIL-2R in patients before and after prednisolone therapy. Methods Serum sIL-2R levels were determined using an immunoenzymatic technique in 97 patients with IgA nephropathy, and the effects of prednisolone therapy on sIL-2R levels were assessed in 15 patients with IgA nephropathy. Results An increased level of serum sIL-2R was shown at the time of renal biopsy in all patients with IgA nephropathy. Moreover, the serum sIL-2R levels were positively correlated with urinary protein excretion (P<0.01,r=0.32) and negatively correlated with creatinine clearance (P<0.01,r=−0.42) in all patients with IgA nephropathy. Prednisolone therapy (0.8 mg/kg per day) caused a significant reduction in the serum sIL-2R level in accordance with the decrease in daily proteinuria in 15 patients with IgA nephropathy. Conclusions These results suggest that the measurement of serum sIL-2R would be useful in evaluating the degree of disease activity, and the serum sIL-2R levels may be a useful indicator for estimating the effect of prednisolone therapy in patients with IgA nephropathy.     

冠状动脉搭桥术后血浆可溶性Fas及其配体的变化

目的比较在心肺转流（CPB）或非CPB下冠状动脉搭桥术（cABG）后可溶性Fas（sFas）和可溶性Fas配体（sFasL）的血浆浓度变化。方法19例病人分别在CPB（CPB组，n=9）或非CPB（非CPB组，n=10）下行择期cABG。术前、术毕和术后取血测定白细胞介素-6（IL-6）、中性粒细胞弹性蛋白酶、sFas、sFasL血浆浓度。结果术毕、术后4h的IL-6和术毕、术后4、12h的中性粒细胞弹性蛋白酶非CPB组明显低于CPB组（P〈0．05或P〈0．01）。CPB组的sFas在术后4、12h明显升高（P〈0．05或P〈0．01），非CPB组在术后12h明显升高（P〈0．01），术后24h恢复至术前水平。两组的sFasL在术毕和术后4、12h均明显升高（P〈0．05或P〈0．01），术后24h恢复至术前水平；其中术后12h非CPB组明显低于CPB组（P〈0．05）。结论CPB或非CPB下行cABG均导致sFas和sFasL血浆浓度升高，但CPB的应用使sFasL血浆浓度升高得更多。sFasL血浆浓度可反映机体炎性反应的程度。     

Fas-induced apoptosis is a feature of progressive diabetic nephropathy in transgenic (mRen-2)27 rats: attenuation with renin-angiotensin blockade

BACKGROUND AND AIM: Tubular atrophy is a major feature of most renal diseases and is closely associated with the loss of renal function. The present study sought to investigate whether Fas/FasL-induced tubular epithelial cell apoptosis was a feature of experimental diabetic nephropathy. The effects of renoprotective therapy with blockade of the renin-angiotensin (RAS) system were also examined. METHOD: Six-week-old female Ren-2 rats were injected with streptozotocin and maintained diabetic for 12 weeks. Further groups of diabetic rats were treated with the angiotensin-converting enzyme inhibitor, perindopril, for 12 weeks. RESULTS: Widespread apoptosis, identified by using mediated Terminal dUTP nick-end labelling (TUNEL) staining was noted in the tubules of diabetic Ren-2 rats. These changes were associated with an increase in both Fas mRNA and Fas L (ligand) within the tubules (P < 0.01). Treatment of diabetic Ren-2 rats with perindopril (6 mg/kg per day) reduced the apoptosis to control levels and was associated with a reduction in Fas mRNA and Fas L protein (P < 0.05). CONCLUSION: In conclusion, Fas/Fas L-induced tubular apoptosis is a feature of diabetic Ren-2 rats and is attenuated by the blockade of the RAS.     

Risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis and diabetic nephropathy: A 3‐year follow‐up study

This study aimed to explore the risk factors for foot ulcer recurrence in patients with comorbid diabetic foot osteomyelitis (DFO) and diabetic nephropathy (DN). This is a prospective cohort study. Between May 2018 and May 2021, we selected 120 inpatients with comorbid severe diabetic foot infection (PEDIS Grade 3 or above) and DN for inclusion in our study. All cases were followed up for 36 months. The study outcomes were whether foot ulcer recurred and the time to recurrence. The risk factors of ulcer recurrence were analysed by comparing the data of the three groups. According to the recurrence of foot ulcer, the participants were divided into three groups: Group A (no foot ulcer recurrence, n = 89), Group B (foot ulcer recurrence within 12‐36 months, n = 19) and Group C (foot ulcer recurrence within 6‐12 months, n = 12). The multivariate Cox regression analysis showed that urine albumin‐creatinine ratio (UACR) (HR: 1.008, 95% CI: 1.005‐1.011, P < .001) and vibration perception threshold (VPT) (HR: 1.064, 95% CI: 1.032‐1.096, P < .001) were identified as independent risk factors. Kaplan‐Meier curves showed a significant positive association between UACR or VPT and the risk of foot ulcer recurrence (log rank, all P < .05). Areas under the ROC curves for UACR, VPT and the combination of UACR and VPT were 0.802, 0.799 and 0.842, respectively. The best cut‐off values of UACR and VPT were 281.51 mg/g and 25.12 V, respectively. In summary, elevated UACR and VPT were independent risk factors. The best clinical cut‐off values of UACR and VPT for prediction of foot ulcer recurrence were 281.51 mg/g and 25.12 V, respectively. Besides, our results suggested that microcirculation disorders rather than macrovascular complications play a major role in the recurrence of foot ulcer in patients with comorbid DFO and DN.     

Examination of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with burns

The FasL–Fas system is one of the recognized apoptosis-inducing systems, and has been determined to have important functions in relation to homeostasis and biological defense mechanisms. In this study, we investigated the serum levels of soluble Fas (sFas), soluble FasL (sFasL) and tumor necrosis factor (TNF-) in patients with burns. The sFas levels were found to be significantly higher in the patients who eventually died as compared to those in the patients who survived (3.9±1.8 ng/ml versus 2.6±1.0 ng/ml). On the other hand, the sFasL levels were significantly higher in the patients who survived (61.5±29.9 ng/ml versus 37.2±14.4 ng/ml) than in those who eventually died. A positive correlation was noted between the TNF- level and the sFas level, and a negative correlation was observed between the TNF- level and the sFasL level. These findings suggest that worsening of the condition of a burns patient may be related to changes in the Fas–FasL system.     

The correlation of inflammatory markers and plasma vaspin levels in patients with diabetic nephropathy

Vaspin, a recently identified adipokine, is a visceral adipose tissue-derived serine protease inhibitor that may have insulin sensitizing effect on adipose tissue. Herein, we measured vaspin level in patients with different stages of diabetic nephropathy (DNP), and investigated the correlation of the vaspin level with other inflammatory parameters. 106 adult type 2 diabetic patients with no known chronic inflammatory disease were included and grouped according to the stage of DNP: Albuminuria <30?mg/day and estimated glomerular filtration rate (eGFR)?>?60?mL/min/1.73m² (Group-1); albuminuria 30–300?mg/day and eGFR >60?mL/min/1.73m² (Group-2); albuminuria >300?mL/min and eGFR <60?mL/min/1.73m² (Group-3). Demographic, clinical and laboratory data were recorded as well as vaspin, high sensitivity C-reactive protein (hsCRP), interleukin (IL)-1 and tumor necrosis factor (TNF)-α levels. There were 38, 35 and 33 patients in Group 1, 2 and 3, respectively. Groups were similar regarding age and gender. Vaspin level did not differ between groups. When all the groups were considered, vaspin was positively correlated with IL-6 level (r?=?0.215, p?=?0.041). No correlation of vaspin was found with IL-1, TNF-α and hsCRP levels (p?=?0.580, r?=?0.054; p?=?0.463, r?=?0.072; p?=?0.812, r?=?0.025, respectively). Vaspin levels of the patients with GFR ≥60?mL/min/1.73m² was less than that of patients with GFR <60?mL/min/1.73m² (p?=?0.03). Age and IL-6 were found to be the major determinants of vaspin level with linear regression analysis. In patients with DNP, vaspin level does not change within the early stages of DNP; while it is higher in patients with decreased GFR, which may be related with increasing inflammation regardless of the stage of the kidney disease.     

Influence of pregnancy on progression of diabetic nephropathy and subsequent requirement of renal replacement therapy in female type I diabetic patients with impaired renal function.

The influence of pregnancy on the progression of diabetic nephropathy in diabetic women with pre-existing moderate renal insufficiency is a subject of considerable controversy in the literature. In four of five female patients with type I diabetes mellitus with pre-existing impaired renal function (creatinine clearance less than 80 ml/min), significant proteinuria (greater than 2 g/24 h urine) and hypertension we have found a further decline in renal function during pregnancy, with an increased deterioration rate of creatinine clearance in comparison to the time before and after pregnancy. The mean decline of the glomerular filtration rate was 1.8 ml/min per month during pregnancy and 1.4 ml/min per month postpartum until the start of dialysis treatment. The difference in the progression of diabetic nephropathy during and after pregnancy can be explained by increased hypertension during pregnancy, especially in the third trimester, despite an intensified antihypertensive therapy. The long-term effect of pregnancy on renal function in our patients was therefore an earlier requirement for renal replacement therapy than would have been expected without pregnancy.     

洛沙坦对糖尿病大鼠肾脏细胞凋亡及Fas和Fas-L表达的影响

目的：探讨血管紧张素Ⅱ1型受体拮抗剂（AT1Ra）对糖尿病大鼠肾脏细胞凋亡及凋亡相关蛋白的影响。方法：单侧肾切除大鼠腹腔注射链脲佐菌素诱发糖尿病模型,每日灌胃给予AT1Ra洛沙坦（40mg/kg),共12周。采用原位末端标记法检测肾脏细胞凋亡情况,流式细胞术和免疫组织化学检测肾皮质Fas和Fas-L表达。结果：糖尿病组较对照组肾小球、肾小管凋亡细胞数明显增多,Fas和Fas-L的表达亦显著增强;洛沙坦治疗组糖尿病组凋亡细胞数减少,Fas和Fas-L的表达减弱。结论：洛沙坦可能通过影响凋亡相关蛋白Fas和Fas-L的表达而抑制肾脏细胞凋亡,从而发挥肾脏保护作用。     

外周血可溶性Fas蛋白及其配体、髓过氧化物酶水平对重症肺炎患儿预后不良的预测价值

目的观察重症肺炎患儿外周血可溶性Fas蛋白（sFas）、可溶性Fas蛋白配体（sFasL）和髓过氧化物酶（MPO）水平变化,并探讨三者对重症肺炎预后不良的预测价值。 方法选取四川中医药高等专科学校绵阳富临医院2016年2月至2020年5月收治的182例重症肺炎、196例轻症肺炎患儿和178例健康儿童,分别为重症组、轻症组和对照组;重症组患儿再根据预后分为预后不良组（29例）和预后良好组（153例）。采用单因素方差分析比较重症组、轻症组治疗前和对照组外周血sFas、sFasL和MPO水平;采用单因素和多因素Logistic回归分析重症组患儿预后不良的影响因素,采用受试者工作特征（ROC）曲线评价外周血sFas、sFasL和MPO水平以及联合指标预测重症组患儿预后不良的价值。 结果三组研究对象的性别、年龄和体重,重症组与轻症组患儿病原微生物分布、肺炎分期差异均无统计学意义（P均> 0.05）。重症组患儿治疗前外周血sFas、sFasL和MPO水平分别为（104.63 ± 19.75）ng/L、（1 062.36 ± 179.85）ng/L和（1 020.26 ± 59.71）U/L,轻症组患儿分别为（80.52 ± 13.66）ng/L、（703.57 ± 127.66）ng/L和（796.75 ± 43.02）U/L,对照组儿童分别为（58.78 ± 10.16）ng/L、（577.83 ± 121.22）ng/L和（632.59 ± 38.71）U/L;重症组和轻症组患儿以上3个指标水平均高于对照组（sFas：重症组 vs.对照组：t = 27.605、P < 0.001;轻症组vs.对照组：t = 17.322、P < 0.001;sFasL：重症组 vs.对照组：t = 29.908、P < 0.001,轻症组vs.对照组：t = 9.744、P < 0.001;MPO：重症组 vs.对照组：t = 71.920、P < 0.001;轻症组vs.对照组：t = 38.647、P < 0.001）,重症组患儿以上3个指标水平均显著高于轻症组（t = 13.885、22.488、41.973,P均< 0.001）。重症组患儿预后不良发生率为15.93%（29/182）。预后不良组患儿双重/多重感染占比（χ² = 12.081、P = 0.001）、多肺叶感染占比（χ² = 32.378、P < 0.001）和外周血白细胞计数（WBC）（t = 6.432、P < 0.001）、中性粒细胞百分比（N%）（t = 3.658、P = 0.001）、C-反应蛋白（CRP）（t = 19.415、P < 0.001）、降钙素原（PCT）（t = 26.101、P < 0.001）、sFas（t = 13.717、P < 0.001）、sFasL（t = 5.357、P < 0.001）和MPO（t = 5.435,P < 0.001）水平均显著高于预后良好组患儿;多因素Logistic回归分析显示以上指标均为重症组患儿预后不良的危险因素,差异均有统计学意义（OR = 5.969、95%CI：4.857~6.304、P = 0.029,OR = 7.485、95%CI：6.785~8.126、P = 0.014,OR = 5.332、95%CI：4.593~5.567、P = 0.010,OR = 4.959、95%CI：4.246~5.337、P = 0.015,OR = 5.143、95%CI：4.879~5.695、P = 0.003,OR = 6.126、95%CI：5.630~6.558、P = 0.008,OR = 8.325、95%CI：6.452~9.902、P = 0.005,OR = 8.469、95%CI：7.879~8.653、P = 0.001,OR = 9.132、95%CI：8.882~9.594,P = 0.003）。外周血sFas、sFasL和MPO水平预测重症组预后不良的Cut-off值分别为125.07 ng/L、1 171.21 ng/L和1 053.04 U/L;sFas、sFasL和MPO以及3个指标联合预测的曲线下面积（AUC）分别为0.875、0.890、0.897和0.955,3个指标联合预测AUC均显著高于sFas、sFasL、MPO水平单独预测,差异均有统计学意义（Z = 5.693、P = 0.005,Z = 5.192、P = 0.007,Z = 4.982、P = 0.009）。 结论重症肺炎患儿外周血sFas、sFasL和MPO水平均偏高,且在预后不良重症患儿中水平均更高,其联合应用可预测患儿不良预后。     

Serum levels of soluble Fas and disease activity in patients with IgA nephropathy

Using a sandwich ELISA, we studied 48 patients with IgA nephropathy and 10 patients with diffuse mesangial proliferative glomerulonephritis without IgA deposition (non-IgA PGN) to determine if levels of serum soluble Fas (s-Fas) might reflect the disease activity. The levels of serum s-Fas in patients with the advanced stage of IgA nephropathy were significantly higher than those in patients with the mild stage of the disease, in non-IgA PGN or in healthy controls. The results showed that advanced stage IgA nephropathy patients who showed heavy proteinuria and the presence of urinary casts revealed high levels of serum s-Fas. It was thus suggested that the measurement of serum s-Fas is useful in evaluating the degree of renal injury in patients with IgA nephropathy.     

2型糖尿病肾病并发骨质疏松大鼠模型研究

目的构建2型糖尿病肾病并发骨质疏松大鼠模型。方法选用SD大鼠,通过喂养高糖高脂饲料4周后,予以链脲佐菌素(STZ 35 mg/kg)进行腹腔注射,继续喂养4周后观察血糖、尿量、尿蛋白及Ca、P等生化指标变化,通过双能X线测量骨密度以及进行生物力学检测骨强度,并取肾、骨组织进行病理检测。结果造模组在高糖高脂饲料喂养后,血糖、尿量及尿蛋白较正常组均逐渐升高,在第4周腹腔注射链脲佐菌素后,造模组的血糖、尿量及尿蛋白较正常组显著升高。在第8周时,造模组的血糖、尿量、尿蛋白及尿Ca、P均明显高于正常组,差异有统计学意义。造模组的骨密度及骨骼强度低于正常组。病理显示造模组大鼠出现肾脏肥大,肾小球毛细血管袢肥大,系膜基质增多肾脏病变以及骨小梁稀疏、变细、连续性中断等骨骼病变。结论通过腹腔注射链脲佐菌素联合8周高糖高脂饮食可建立2型糖尿病肾病并发骨质疏松的大鼠模型,该模型具有高糖、多尿、尿蛋白增加、骨密度下降以及肾小球病变和骨吸收增加的特点。     

The predictive value of diabetic retinopathy on subsequent diabetic nephropathy in patients with type 2 diabetes: a systematic review and meta-analysis of prospective studies

This systematic review and meta-analysis aimed to assess the predictive value of diabetic retinopathy (DR) on further diabetic nephropathy (DN) risk in patients with type 2 diabetes (T2D) based on the prospective cohort studies. PubMed, Embase, and the Cochrane Library were systematically searched for eligible prospective cohort studies through March 2020. The predictive value of DR was assessed using sensitivity, specificity, positive likelihood ratio (PLR) and negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and area under the receiver operating characteristic curve (AUC) through the bivariate generalized linear mixed model and the random-effects model. Ten prospective cohort studies recruited 635 patients with T2D. The pooled sensitivity and specificity of DR for predicted DN were noted to be 0.64 (95% CI, 0.54–0.73) and 0.77 (95% CI, 0.60–0.88), respectively. The pooled PLR and NLR of DR for predicted DN were 2.72 (95% CI, 1.42–5.19) and 0.47 (95% CI, 0.33–0.67), respectively. The summary DOR for the relationship between DR and subsequent DN for T2D patients was 5.53 (95% CI, 2.00–15.30), and the AUC of DR for predicted DN was 0.73 (95% CI, 0.69–0.77). This study found significant associations between DR and subsequent DN risk for patients with T2D. Moreover, the predictive value of DR on subsequent DN risk was relatively lower.     

BPH患者膀胱逼尿肌细胞凋亡及Fas、FasL 表达的研究

目的观察BPH膀胱逼尿肌平滑肌细胞的凋亡和凋亡相关基因Fas、FasL的表达及它们之间的相互关系.方法对25例BPH患者和15例同龄无膀胱流出道梗阻(BOO)患者的膀胱逼尿肌,用末端脱氧核糖核酸转移酶(TUNEL)法检测凋亡指数,免疫组化方法检测Fas、FasL蛋白的表达水平.并用光镜和透射电镜观察了逼尿肌结构改变,电镜下的细胞凋亡情况.结果BPH组膀胱逼尿肌平滑肌细胞凋亡指数(AI)为19.07±4.52,较对照组8.57±3.20显著升高(P＜0.01).BPH组Fas的阳性表达率为96.0%,较对照组73.3%明显增加(P＜0.01).BPH组FasL的阳性表达率为96.0%,较对照组66.7%明显增加.(P＜0.01).光镜和电镜下BPH组逼尿肌细胞形态紊乱,细胞问结缔组织增生,电镜下BPH组凋亡细胞相对增多.结论BPH患者逼尿肌较同龄正常逼尿肌细胞凋亡率上升,凋亡基因Fas和FasL的表达升高,提示此改变是引起凋亡上升的原因.除去超微结构的改变等影响了BPH患者膀胱逼尿肌功能外,逼尿肌细胞凋亡增加也可能是导致逼尿肌功能减退的原因.     

Effect of activin A on tubulointerstitial fibrosis in diabetic nephropathy

Aim:   The effect of activin A on tubulointerstitial fibrosis in diabetic nephropathy (DN) using streptozotocin (STZ)-induced diabetic rats and high glucose-cultured HK-2 cells was investigated.
Methods:   Male Wistar rats were randomized into a normal control group (NC) and diabetes mellitus group (DM). Diabetes was induced by i.p. injection of STZ. Six rats were respectively killed 4, 8, 12 and 16 weeks after model establishment in each group. The changes of kidney weight/bodyweight (KW/BW), urine albumin excretion rate (AER) and creatinine clearance rate (Ccr) were determined. The morphology of tubulointerstitium was observed by light microscopy. Further biochemical analysis was provided using immunohistochemistry and real-time polymerase chain reaction. The different parameters in high glucose-cultured HK-2 cells were monitored by western blotting or enzyme-linked immunosorbent assay (ELISA) and the intervention of rh-follistatin on them was investigated.
Results:   Compared with the NC group, there was marked enlargement in the levels of KW/BW, AER, Ccr and interstitial fibrosis index, and the production of P-Smad2/3 and fibronectin in the DM group from 8 to 16 weeks. Activin βA, mainly located in tubular epithelial cells, was significantly higher in the DM group than that in the NC group throughout the study periods. Follistatin was abundant in the NC group, but was diminished gradually in the DM group. High glucose may facilitate the synthesis of activin βA, transforming growth factor (TGF)-β, P-Smad2/3 and fibronectin in HK-2 cells while rh-follistatin inhibited them except TGF-β.
Conclusion:   Activin A is involved in tubulointerstitial fibrosis in DN by inducing the production of fibronectin through Smad signal pathway.     

Fas基因在胰腺癌细胞凋亡中的研究

目的 探讨雌激素受体拮抗剂三苯氧胺诱导雌激素受体阳性胰腺癌细胞凋亡过程中与Fas基因间的关系。方法 流式细胞仪分离：Fas^( )和：Fas^(-)胰腺癌细胞，通过TAM诱导Fas^( )和Fas^(-)胰腺癌细胞株Bxpc-3凋亡，MTT法检测细胞活度，TUNEL法检测细胞凋亡，采用RT-PCR-ELISA法检测端粒酶活性，进行相关性分析。结果 TAM可以诱导出Fas^( )胰腺癌细胞凋亡，不能诱导出Fas^(-)胰腺癌细胞凋亡。结论 TAM诱导雌激素受体阳性胰腺癌细胞株Bxpc-3凋亡，必须有Fas^( )抗体的介导，不能单纯用表面膜受体拮抗剂的理论来解释TAM诱导雌激素受体阳性胰腺癌细胞株Bxpc-3凋亡。     

Disparate phospho-Smad2 levels in advanced type 2 diabetes patients with diabetic nephropathy and early experimental db/db mouse model

Uncontrolled activation of transforming growth factor beta (TGF-β) family members is hypothesized to participate in type 2 diabetes (T2D) dependent diabetic nephropathy (DN). We evaluated and compared downstream activation of the Smad2-signaling pathway in kidney samples from T2D patients to kidneys from the T2D model of leptin receptor deficient db/db mouse. Furthermore, expression of TGF-β family members was evaluated to elucidate molecular mechanisms in the mouse model. Kidney samples from patients with advanced stages of DN showed elevated pSmad2 staining whereas db/db mouse kidneys surprisingly showed a decrease in pSmad2 in the tubular compartment. Structurally, kidney tissue showed dilated tubules and expanded glomeruli, but no clear fibrotic pattern was found in the diabetic mice. Selective TGF-β family members were up-regulated at the mRNA level. Antagonists of bone morphogenetic protein (BMP) ligands, such as Gremlin1, USAG1 and Sclerostin, were strongly up-regulated suggesting a dampening effect on BMP pathways. Together, these results indicate a lack of translation from T2D patient kidneys to the db/db model with regards to Smad signaling pathway. It is plausible that a strong up-regulation of BMP antagonizing factors account for the lack of Smad1/5/8 activation, in spite of increased expression of several BMP members.     

厄贝沙坦对糖尿病肾病大鼠病理改变及足细胞凋亡的影响

目的 观察厄贝沙坦对糖尿病肾病大鼠病理改变及足细胞凋亡情况的影响,探讨厄贝沙坦对糖尿病肾损伤的保护作用机制.方法 腹腔注射链脲佐菌素建立糖尿病肾病大鼠模型.将36只雄性Wistar清洁级健康大鼠按随机数字表法分为正常对照组(N组)、糖尿病肾病组(DN组)和厄贝沙坦干预组(E组),于用药第12周取肾组织行苏木素-伊红染色及PAS染色,光镜下观察各组肾脏病理改变情况,采用TUNEL法计数凋亡足细胞数目.结果 12周时,DM组、E组大鼠24 h尿蛋白定量与N组比较差异有统计学意义(P＜0.01);E组大鼠24 h尿蛋白定量与DM组比较差异有统计学意义(P＜0.01);光镜下各组肾脏病理改变显示,N组大鼠肾小球形态未见异常变化,采用TUNEL法观察足细胞形态结构正常,未见凋亡足细胞;DN组大鼠肾脏体积较N组增大,系膜区增宽,可见不同程度的系膜细胞增生;采用TUNEL法显示足细胞体积肿胀增大,凋亡足细胞数目明显增多(P＜0.01);E组大鼠肾小球毛细血管基底膜增厚明显改善,凋亡足细胞数目较DM组明显减少(P＜0.01).结论 厄贝沙坦对糖尿病肾病的保护作用机制可能是通过改善肾小球滤过膜结构、抑制足细胞凋亡机制来减少糖尿病肾病大鼠大量蛋白尿,改善糖尿病肾病的预后.

Abstract：

Objective To investigate the influnce of irbesartan on the structure of glomerular filtration memberance and apoptosis of podocytes in diabetic nephropathy (DN) rats.Methods Give intraperitoneal injection of streptozotocin(STZ) to establish models of DN rats. 36 male Wistar clean rats were divided the into 3 groups,Normal Comparison group (N group), diabetic nephropathy group (DN group) and treatment group with Irbesartan (50 mg/kg. d E group) . After 12 weeks we observe 24 h urinary Protein count and the pathological changes of the kidney by extracting renal tissue to do HE staining and PAS staining; to count the apoptosis of podocytes of rats using TUNEL method. Results the level of 24 hour Protein in group E and group DM increased compared with those in the group N.,while the level of 24 hour Protein in group E decreased compared with that in the group DN after 12 weeks (P ＜ 0.01). For N group rats: glomerulus shapes are normal by optical microscopes, the structures of the podocytes are normal, using TUNEL method and there is no apoptotic podocytes. For DM group rates, the volume of kidney is bigger than N group under optical microscope, mesangial region becomes wider and there is different degrees of mesangial hyperplasia. Apoptotic podocytes increased than that of N group( P ＜ 0.01). As for the E group, basement membrane of glomerulus capillary improved significantly, the number of apoptotic podocytes decreased noticeably compared with those of the DM group (P ＜ 0.01). Conclusion The protection mechanism of the irbesartan to the diabetic nephropathy (DN) is probably through the improvement of the structure of glomerular filtration memberance, refraining the mechanism of the apoptosis of podocytes.