摘要：
目的 初步探讨雷公藤甲素对C57BL/6－Ins2Akita小鼠的安全剂量和延缓糖尿病肾病的作用效果.方法 选择8周龄雄性C57BL/6－Ins2Akita小鼠和C57BL/6J野生鼠,每4周检测体血糖和体质量,每8周收集24 h尿液和血液,检测和对比糖尿病肾病相关指标,确定C57BL/6－Ins2Akita小鼠糖尿病肾病的评估和形成时机.另取8周龄C57BL/6－Ins2Akita小鼠以100、200、400、800 μg· kg－1· d－1分成4个剂量组灌胃14 d,观察急性毒性反应,通过改良寇氏法计算 LD50.以 25、50、100 μg· kg－1· d－1这 3 个剂量干预 24 周龄C57BL/6－Ins2Akita小鼠,并设置空白对照组和野生型对照组,干预8周,期间收集尿液检测24 h尿蛋白排泄率,处死后收集血液检测肌酐、尿素氮、血清清蛋白等指标,组织取材进行病理检查.结果 C57BL/6－Ins2Akita小鼠的24 h尿蛋白排泄率、血糖、三酰甘油在8周龄开始即高于C57BL/6J野生型小鼠(P＜0.05),血清蛋白低于C57BL/6J野生型小鼠(P＜0.05),且随周龄增加差距逐渐明显,但肌酐未出现升高趋势.急性毒性实验中改良寇氏法计算LD50值为356.45 μg· kg－1· d－1.雷公藤甲素干预高、中、低剂量组的24 h尿清蛋白排泄率均较对照降低,其中高剂量组与对照组差异有统计学意义(P＜0.05).随蛋白尿的减少,血清清蛋白水平得到提高, HDG组较CON组血清清蛋白水平提高约2.5 g/L,差异有统计学意义(P＜0.05).对于体质量、24 h尿量、肾脏重量、血糖、血肌酐、尿素氮、三酰甘油、胆固醇等指标,实验组与空白对照组之间未发现差异有统计学意义.与此同时,病理检查发现高剂量组的系膜面积/毛细血管襻面积和肾小球系膜扩张比例均明显低于空白对照组(P＜0.05),中、低剂量组差异无统计学意义.结论 100 μg· kg－1· d－1的雷公藤甲素可减少C57BL/6－Ins2Akita小鼠的蛋白尿,提高血清蛋白水平,减少系膜区系膜基质的聚积,增大剂量可能引起毒性反应.%Objective To investigate the safe dosage of triptolide for C 57BL/6－Ins2Akita mice and its effect on delaying the progression of diabetic nephropathy.Methods The male C57BL/6－Ins2Akita mice and C57BL/6J mice were selected in this experient ,From 8weeks old they were examed blood glucose ,blood pressure every 4－5 weeks,24 hour urine albumin,serum creatinine,urea,lipids and liver function were checked every 8 weeks,in order to determine the time for the occurrence and assessment of diabetic nephropathy .At the same time,we intragastrically administrated 8－week－age C57BL/6－Ins2Akita mice with 100,200,400,800 μg· kg－1· d－1for 14 days.Observing the acute toxic reac－tions and calculating the LD50by bliss.Based on above experiments and references ,we decided to intervene 24－week－age C57BL/6－Ins2Akita mice with 25,50,100 μg· kg－1· d－1for 8 weeks.And set the blank control and wild control groups.Meanwhile,we collect urine to detect the urinary albumin excretion rate for 24 h,and urinary protein/creatinine, following by collecting the blood to detect some indexes such as the creatinine ,urea nitrogen and serum albumin after killing the mice.Results The level of urinary albumin excretion rate , blood sugar, triglyceride and Cholesterol in C57BL/6－Ins2Akita mice were higher than those in C57BL/6J mice at all ages(P＜0.05),while the level of serum al－bumin was lower than that of C 57BL/6J wild mice.The gap was gradually drawn apart with age increasing.However,the creatinine did not show the ascendant tendency.The value of LD50is 356.45 μg· kg－1· d－1.Compared with the con－trol,the high,medium and low triptolide experiment groups have less urinary albumin excretion rate with a statistically significant difference in high dose group ( P＜0.05).Serum albumin increased following the reduction of albuminuria. The serum albumin of HDG group increased by 2.5 g/L compared with CON group ,which is statistically significant (P＜0.05).There was no statistically significant difference in body weight ,urine amount for 24h,kidney weight,blood sugar, blood creatinine,urea nitrogen,triglyceride and cholesterol etc between experiment and control group.Meanwhile,me－sangial area/capillary loop area as well as the glomerular mesangial membrane expansion rate in high dose group are ob －viously under the blank control group(P＜0.05)while no statistically significant difference was observed in low and me－dium groups.Conclusion Within safe dosages,high dose triptolide can reduce the albuminuria ,alleviate accumulation of mesangial matrix and increase blood albumin in C57BL/6－Ins2Akita mice,toxic reactions are prone to occur with higher doses.
