摘要目的 观察TDZD-8干预大鼠重症急性胰腺炎(SAP)肾损伤的量效关系,并将三种常用GSK-3β 抑制剂TDZD-8、氯化锂(LiCL)、SB216763对该模型的作用效果进行对比,以探讨针对SAP并发肾损伤大鼠模型最有效的GSK-3β 抑制剂类别及其有效、安全的最佳剂量.方法 96只SPF级雄性Wistar大鼠,随机(随机数字法)分为8组(n=12):假手术组(SO组)、重症急性胰腺炎组(SAP组)、TDZD-8预处理组0.25、0.5、1.0、2.0 mg/kg(TD组,分别标记为TD1、TD2、TD3、TD4组),LiCL预处理组(L组)和SB216763预处理组(SB组),胰胆管逆行注射5％ 牛磺胆酸钠制作SAP模型.术后12 h剖杀各组大鼠,测定各组大鼠腹水量、血清AMY、Cr、BUN和ALT水平,并观察胰腺、肾脏组织病理学变化.结果 SAP组腹水量、AMY、Cr、BUN、ALT水平以及胰腺、肾脏病理评分均较SO组显著升高(P<0.05);TD1组几乎对SAP无缓解作用,TD2、TD3、TD4、L组和SB组均能不同程度地减少SAP大鼠的腹水量,降低血清AMY、Cr、BUN水平,并显著降低胰腺组织病理评分,差异有统计学意义(P<0.05);TD2、TD3均能不同程度地减少ALT水平(P<0.05),而TD4组ALT水平较高,与SAP组相似;TD2对各个指标的效果不如TD3组作用显著,两者比较各项指标差异均有统计学意义(P<0.05).对SAP大鼠腹水量和ALT水平的改善作用,TD组中最佳剂量组TD3组与L组和SB组比较,差异无统计学意义(P>0.05);而TD3组的AMY、Cr、BUN水平以及胰腺、肾脏病理评分均较L组和SB组降低更显著,差异有统计学意义(P<0.05);L组Cr、BUN水平和胰腺、肾脏病理学评分与SB组比较均降低更显著,差异有统计学意义(P<0.05);Western blot检测发现各组中GSK-3β 蛋白表达均处于较一致的水平,差异无统计学意义(P>0.05);而p-GSK-3βser9在SAP组表达低于SO组,差异有统计学意义(P<0.05);TD3、L、SB三组中的p-GSK-3βser9均较SAP组明显增强,其中以TD3组表达最强,同时L组表达强于SB组,组间比较差异均有统计学意义(P<0.05).结论 通过对TDZD-8、LiCL和SB216763对大鼠SAP肾损伤模型的作用比较可知TDZD-8是针对该模型最有效的GSK-3β 抑制剂.对于牛磺胆酸钠诱导的SAP并发肾损伤大鼠模型,静脉给予TDZD-81 mg/kg预处理对该模型是安全有效的最佳剂量.

abstractsObjective To observe the dose-response relationship of the GSK-3β inhibitor TDZD-8 in severe acute pancreatitis (SAP) associated kidney injury in rats. In order to identify the most effective class of GSK-3β inhibitor and its effective and reasonable safe dose in SAP associated kidney injury model in rats by comparing three kinds of frequently-used GSK-3β inhibitor TDZD-8, lithium chloride (LiCL), SB216763 in this model. Methods Totally 96 SPF male Wistar rats were randomly(random number) divided into 8 groups (n=12): sham operation group (SO group), severe acute pancreatitis group (SAP group), TDZD-8 pretreatment groups (TD group, marked TD1, TD2, TD3 and TD4 group, respectively) at different dosage (0.25, 0.5, 1.0 and 2.0 mg/kg), LiCL pretreatment groups (L group, 40 mg/kg), and SB216763 pretreatment group (SB group, 1 mg/kg). SAP model was induced by retrograde infusion of 5％ sodium taurocholate into the biliopancreatic duct. Rats in each group were sacrificed at 12 h after operation. Then the mortality, quantity of ascites, serum AMY, Cr, BUN and ALT were recorded, and the pathological changes of pancreatic tissues and kidney tissues were observed. Results Compared with the SO group, the levels of ascites, serum AMY, Cr, BUN, ALT and pancreatic and renal pathologic score in the SAP group were all significantly increased (P<0.05). Compared with the TD1 group, quantity of ascites, serum AMY, Cr, BUN,ALT and pancreatic tissue pathological grading were reduced in different degrees in the TD2, TD3 and TD4 groups with statistically significant difference (P<0.05); ALT values were reduce in different degrees in the TD2 and TD3 groups as compared with the SAP group (P<0.05), while ALT value in the TD4 group was similar to that in the SAP group; compared with the TD2 group, all the indexes in the TD3 group were significant better (P<0.05); Compared with TD3 group (the best group in TD group), the levels of ascites and serum ALT in the L group and SB group had no significant difference (P>0.05), but the levels of AMY, Cr, BUN, ALT, pancreatic and renal pathologic score were significantly reduced in the TD3 group than those in the L and SB groups (P<0.05); compared with the SB group, the values of Cr, BUN, pancreatic and renal pathologic score in the L group were lower (P<0.05). GSK-3βprotein expression in all groups showed no obvious difference (P>0.05), while p-GSK-3β ser9 protein expression in the SAP group was lower than that in the SO group (P<0.05), and p-GSK-3β ser9 protein expression in the TD3, L and SB groups were stronger than that in the SAP group. Among them, p-GSK-3βser9 protein expression was highest in the TD3 group, followed by the L group, finally the SB group, and the differences were statistically significant (P<0.05). Conclusions Among the three different GSK-3βinhibitors, TDZD-8 is the most effective GSK-3β inhibitor for SAP associated with kidney injury in rats. The GSK-3β inhibitor TDZD-81 mg/kg administered intravenously is safe, effective and optimal dosage for attenuating the severity of severe acute pancreatitis associated with kidney injury.