冬虫夏草预防糖尿病大鼠对比剂肾病机制研究

目的探讨冬虫夏草预防糖尿病大鼠对比剂肾病的机制。方法SD大鼠80只,腹腔注射链脲佐菌素(60 mg/kg)建立糖尿病模型,并随机分为对照组(C组)、对比剂肾病组(CIN组)、冬虫夏草低剂量组(L组)、冬虫夏草标准剂量组(M组)和冬虫夏草强化剂量组(H组)。L组、M组、H组大鼠分别给予冬虫夏草(2.0、2.5、3.0 g/kg)灌胃,C组、CIN组则以等量生理盐水灌饲,连续7天。7天后检测SCr、BUN、尿蛋白排泄率(UAE)、一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)、中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、肾损伤分子1(KIM-1)及IL-18。取左肾测病理和氧化应激指标;右肾用于Western Blot检测抗凋亡蛋白和促凋亡蛋白(包括:Bcl-2、Bax、Caspase-3、Caspase-9、p-Akt、p-mTOR、p-ERK、p-JNK、p-P70S6K)表达。结果与C组比较,CIN组SCr、BUN、UAE、MDA、NAGL、KIM-1、IL-18均明显升高,NO、T-SOD降低(P<0.05)。与CIN组比较,L组、M组、H组SCr、BUN、UAE、MDA均明显降低,NO、T-SOD升高(P<0.05);L组、H组NAGL、IL-18、降低(P<0.01),其中H组KIM-1均明显降低(P<0.01)。与L组、M组比较,H组SCr、BUN、UAE、MDA、NGAL、KIM-1、IL-18明显降低,NO、T-SOD升高(P<0.05)。与C组比较,CIN组Bcl-2、P-Akt、P-mTOR、P-P70S6K、P-ERK蛋白表达增加,Bax、P-JNK、Caspase-3、Caspase-9蛋白表达减少(P<0.05)。与CIN组比较,Bcl-2、P-Akt、P-mTOR、P-P70S6K、P-ERK蛋白表达在L组、M组、H组逐渐增加,其中H组最高(P<0.05),Bax、P-JNK、Caspase-3、Caspase-9蛋白表达L组、M组、H组逐渐减少,H组表达量最少(P<0.05)。结论冬虫夏草通过抗氧化应激、抑制脂质过氧化反应、抗凋亡等途径对糖尿病大鼠对比剂肾病发挥较好的保护作用,冬虫夏草强化剂量组优于低剂量组和标准剂量组。

Study on the Mechanism of Cordyceps Sinensis in Preventing Diabetic Rats with Contrast Induced Nephropathy

Objective To investigate the mechanism of cordyceps sinensis in preventing contrast induced nephropathy(CIN)in diabetic rats.Methods Totally 80 SD rats were intraperitoneal injected with streptozotocin(60 mg/kg)to establish a diabetes model and were randomly divided into control group(group C),CIN group,cordyceps sinensis low dose group(group L),cordyceps sinensis standard dose group(group M),and cordyceps sinensis enhanced dose group(group H).The rats in group L,group M and group H were administered with cordyceps sinensis(2.0,2.5 and 3.0 g/kg)by gavage,while the rats in group C and CIN group were given the equal volume of normal saline for 7 consecutive days.7 days later,the level of SCr,BUN,urinary albamin excretion rate(UAE),nitric oxide(NO),superoxide dismutase(SOD),malondialdehyde(MDA),total antioxidant capacity(T-AOC),neutrophil gelatinase-associated lipocalin(NGAL),kidney injury molecule(KIM-1)and IL-18 were detected.Pathological and oxidative stress indexes were measured in the left kidney.The right kidney was used to detect the expression levels of anti-apoptotic and pro-apoptotic proteins(including Bcl-2,Bax,Caspase-3,Caspase-9,p-AKT,P-mTOR,P-ERK,P-JNK,p-p70s6k)by Western Blot.Results Compared with group C,the contents of SCr,BUN,UAE,MDA,NAGL,KIM-1 and Il-18 in CIN group were significantly increased,while the contents of NO and T-SOD were decreased(P<0.05).Compared with CIN group,the contents of SCr,BUN,UAE and MDA in group L,M and H were significantly decreased,while the contents of NO and T-SOD were increased(P<0.05).The values of NAGL and IL-18 in group L and H were significantly lower(P<0.01),and the values of KIM-1 in group H were significantly lower(P<0.01).Compared with group L and M,SCr,BUN,UAE,MDA,NGAL,KIM-1 and Il-18 levels in group H were significantly decreased,while NO and T-SOD levels were increased(P<0.05).Compared with group C,the expression of Bcl-2,P-AKT,P-mTOR,P-P70S6K and P-ERK increased in CIN group,while the expression of Bax,p-JNK,Caspase-3 and Caspase-9 decreased(P<0.05).Compared with CIN group,the protein expressions of Bcl-2,P-AKT,P-mTOR,P-P70S6K and P-ERK increased gradually in group L,M and H,with the highest expression in group H(P<0.05),and the protein expressions of Bax,p-JNK,Caspase-3 and Caspase-9 gradually decreased in group L,M and H,with the lowest expression in group H(P<0.05).Conclusion Cordyceps sinensis has a better protective effect on diabetic rats with CIN by anti-oxidative stress,inhibition of lipid peroxidation,and anti-apoptosis,and the enhanced dose group of cordyceps sinensis is better than the low-dose group and the standard dose group.