冬虫夏草对他克莫司诱导肾脏和胰腺损伤的保护作用

发布时间：2018-04-24 19:23

本文选题：冬虫夏草 + 他克莫司　；参考：《延边大学》2017年硕士论文

【摘要】：目的探讨冬虫夏草(Cordyceps sinensis,CS)对他克莫司(Tacrolimus,TAC)诱导的大鼠肾及胰腺损伤的保护作用及其机制。方法36只雄性SD大鼠随机分为4组(每组9只),正常对照组(VH):0.25 mL/kg/d橄榄油皮下注射和生理盐水1ml/kg/d灌胃;CS组:CS 5g/kg/d灌胃;TAC组:TAC 1.5mg/kg/d皮下注射和生理盐水1ml/kg/d灌胃;TAC+CS组:TAC1.5mg/kg/d皮下注射,CS5g/kg/d灌胃,各组4周。4周后利用糖耐量实验(An intraperitoneal Glucose tolerance test,IPGTT)检测大鼠血糖,糖促胰岛素分泌实验(Glucose-stimulated insulin secretion,GSIS)检测胰岛素浓度。利用免疫组化法检测胰岛大小。采用ELISA法测定各组大鼠血浆尿8-轻基脱氧鸟苷(8-hydroxy-2'-deoxyguanosine,8-OHdG)的表达和尿8-OHdG排泄率,应用免疫组化法检测肾组织中骨桥蛋白(Osteopontin,OPN)、外胚层发育不良抗体(Ectodermal Dysplasial,ED-1)、'肾母细胞瘤蛋白抗体(Wilms Tumor protein,WT1)、锰超氧化物歧化酶(Manganese-containingsu2 peroxidedismutase,MnSOD)和含半胱氨酸的天冬酸蛋白水解酶 3(Cysteinyl aspartate specific proteinase,Caspase-3)的表达;Masson三色染色法观察肾间质纤维化程度(Tubulointerstitial fibrosis,TIF),应用免疫印迹法检测MnSOD和Caspase-3的表达,采用TUNEL染色法观察肾组织中的凋亡细胞。结果1.与VH组相比,TAC组大鼠的体重明显降低,24小时尿量及摄水量、血肌酐和尿素氮均升高(p0.05),与TAC组相比,CS+TAC组大鼠体重增加,血肌酐和尿素氮均降低(p0.05)。2.与VH相比,TAC组血糖明显升高,血浆胰岛素浓度和胰岛大小均明显降低;与TAC组相比,CS+TAC组胰岛功能明显改善(均p0.05)。3.与VH组相比,TAC组TIF和WT1表达明显升高;与TAC组相比,CS+TAC组 TIF(10.8 ± 2.4 vs 5.2± 0.6)和 WT1表达明显下降(15.8±0.6 vs 10.3± 0.9,p0.05)。4.与VH组相比,TAC组大鼠肾组织中OPN和ED-1表达明显升高;与TAC组相比,CS+TAC组OPN(9.3±0.8 vs 3.4±0.3)和 ED-1表达明显降低(23.1±2.5 vs 14.8 ± 1.8,p0.05)。5.与VH组相比,TAC组尿8-OHdG排泄率增加,血清8-OHdG含量增加;与 TAC 组相比,CS+TAC 组尿 8-OHdG(220.0 ± 12.0ng/d vs 145.2± 14.2 ng/d)和血清 8-OHdG(3±0.06ng/ml vs 1.34±0.05ng/ml,均p0.05)均显著降低。与VH组相比,TAC组MnSOD表达降低,与TAC组相比,CS+TAC组MnSOD(1.3±0.5 vs 5.6±0.7,p0.05)表达升高。6.与VH组相比,TAC组Caspase-3的表达增高、TUNEL阳性细胞数增高,与TAC组相比,CS+TAC组两者表达均明显下降(Caspase-3:8.9± 0.8 vs 3.6±0.6,TUNEL:11.6±2.4cell/mm2 vs 5.2± 1.2cell/mm2,p0.05)。结论1.冬虫夏草对他克莫司诱导胰腺损伤的保护作用,其机制与冬虫夏草改善胰岛β细胞功能相关。2.冬虫夏草对他克莫司诱导肾脏损伤的保护作用,其机制与冬虫夏草抗炎、抗纤维化、抗凋亡、抗氧化应激、减轻肾小球足细胞损伤有关。
[Abstract]:Objective to investigate the protective effect of Cordyceps sinensis CSC on renal and pancreatic injury induced by tacrolimus in rats. Methods Thirty-six male Sprague-Dawley rats were randomly divided into 4 groups (n = 9 in each group), normal control group (n = 9), normal control group (n = 9) and control group (n = 9). The control group was given subcutaneous injection of VH: 0.25 mL/kg/d olive oil and normal saline 1ml/kg/d respectively. The rats in the control group were given subcutaneous injection of mL/kg/d 1.5mg/kg/d of 1: CS 5g/kg/d and 1. 5 mg / kg / d of CS 5 mg / kg 路d in the control group and the normal saline 1ml/kg/d group (n = 1. 5 mg / kg / d). After 4 weeks, glucose tolerance test (an intraperitoneal tolerance test) and glucose stimulating insulin (Glucose-stimulated insulin secretion) were used to detect blood glucose and insulin concentration in rats. The size of islets was detected by immunohistochemical method. The expression of 8-hydroxy-2-deoxyguanosine 8-hydroxy-2-deoxyguanosine 8-OHdG and urinary 8-OHdG excretion rate were measured by ELISA assay. Immunohistochemical method was used to detect osteopontin, ectodermal dysplasia antibody (Ectodermal dysplasia), Wilms Tumor protein (WT1), manganese superoxide dismutase (Manganese-containing su2 peroxidedismutase) and 3(Cysteinyl aspartate specific in renal tissues. The expression of Caspase-3 in renal interstitial fibrosis was observed by Masson trichrome staining, and the expression of MnSOD and Caspase-3 were detected by Western blot. Apoptotic cells in renal tissue were observed by TUNEL staining. Result 1. Compared with the VH group, the body weight of the rats in the TAC group decreased significantly, the urine volume and water intake decreased significantly, and the levels of serum creatinine and urea nitrogen increased (P 0.05). Compared with the TAC group, the weight of the rats in the CS TAC group increased, and the levels of serum creatinine and urea nitrogen decreased. Compared with the VH group, the plasma insulin concentration and islet size were significantly decreased, and the islet function in the CS TAC group was significantly improved compared with the VH group (all p 0.05. 3). Compared with the VH group, the expression of TIF and WT1 was significantly increased, the expression of TIF(10.8 卤2.4 vs 5.2 卤0.6) and the expression of WT1 decreased significantly (15.8 卤0.6 vs 10.3 卤0.9 p0.05 路4) in the CS TAC group compared with the VH group, and the expression of TIF(10.8 卤2. 4 vs 5.2 卤0. 6 in the CS TAC group was significantly lower than that in the VH group. Compared with the VH group, the expression of OPN and ED-1 in the renal tissue was significantly increased, and the expression of OPN(9.3 卤0.8 vs 3.4 卤0.3 in the CS TAC group was significantly lower than that in the TAC group, and the expression of ED-1 was significantly lower than that in the VH group (23.1 卤2.5 vs 14.8 卤1.8 p0.05 路5). Compared with VH group, urinary 8-OHdG excretion rate increased and serum 8-OHdG content increased, and serum 8-OHdG(220.0 卤12.0ng/d vs 145.2 卤14.2ng / d in CS TAC group and 8-OHdG(3 卤0.06ng/ml vs 1.34 卤0.05ng / ml (p0.05) in CS TAC group were significantly lower than those in VH group. Compared with the VH group, the expression of MnSOD was decreased in the MnSOD group, and the MnSOD(1.3 卤0.5 vs 5.6 卤0.7 p0.05) expression in the CS TAC group was significantly higher than that in the TAC group. Compared with VH group, the expression of Caspase-3 was increased and the number of Tunel positive cells was increased. Compared with TAC group, the expression of Caspase-3: 3: 8. 9 卤0. 8 vs 3.6 卤0. 6% Tunel: 11.6 卤2.4cell/mm2 vs 5.2 卤1.2? Conclusion 1. The protective effect of Cordyceps sinensis on pancreatic injury induced by tacrolimus was related to the improvement of islet 尾-cell function by Cordyceps sinensis. The protective effect of Cordyceps sinensis on the renal injury induced by tacrolimus is related to its anti-inflammatory, anti-fibrosis, anti-apoptosis, anti-oxidative stress and reducing the damage of glomerular podocytes.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R979.5

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