丹参酮ⅡA联合DAPT对UUO模型大鼠肾组织 Notch/Jagged通路相关蛋白表达的影响

Effect of TanshinoneⅡA Combined with DAPT on Notch/Jagged Pathway Proteins in Renal Tissue of UUO Rat Models

目的观察丹参酮ⅡA配伍γ-分泌酶抑制剂3,5-二氟苯乙酰-L-丙氨酰-S-苯基甘氨酸-t-丁酯(DAPT)对单侧输尿管梗阻(UUO)模型大鼠肾组织Notch/Jagged通路相关蛋白NICD、Hey1、Rbp-Jk及ColⅠ表达的影响,探讨其防治肾纤维化的分子机制。方法取24只大鼠结扎右侧输尿管建立UUO模型,随机分为模型组、丹参酮ⅡA组、DAPT组、配伍组,每组6只。另取6只大鼠设为正常组。正常组及模型组给予羧甲基纤维素钠(CMC-Na)灌胃,其余各给药组分别给予丹参酮ⅡA(25 mg/kg)、DAPT(12 mg/kg)、丹参酮ⅡA(25 mg/kg)+DAPT(12 mg/kg)灌胃,2周后处死大鼠取右肾组织。采用Masson染色观察肾组织胶原纤维,免疫组织化学法检测各组大鼠肾组织NICD、Hey1表达,Western Blot技术检测各组大鼠肾组织Rbp-Jk、ColⅠ表达。结果与正常组比较,模型组Masson染色评分升高(P<0.05),NICD、Hey1、Rbp-Jk、ColⅠ蛋白表达均升高(P<0.05)。与模型组比较,各给药组Masson染色评分、NICD、Hey1、Rbp-Jk、ColⅠ蛋白表达均降低(P<0.05)。与丹参酮ⅡA组及DAPT组比较,配伍组Masson评分、NICD、Hey1、Rbp-Jk、ColⅠ蛋白表达均降低(P<0.05)。结论丹参酮ⅡA配伍DAPT可改善UUO模型肾间质病理损害,减轻肾纤维化过程中胶原纤维合成沉积,其机制可能与抑制NICD、Rbp-Jk、Hey1及ColⅠ在肾组织中的表达,影响Notch/Jagged信号途径转导相关。

Objective To observe the effects of TanshinoneⅡA combined with γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-L-alanyl]-s-phenylglycine t-butyl ester(DAPT) on the protein expressions of Notch/Jagged pathway, i.e., NICD, Hey1, Rbp-Jk and ColⅠ in renal tissue of UUO rat models, and to explore it′s molecular mechanism in preventing and treating renal fibrosis. MethodsTwenty-four rats were recruited for establishing UUO models by ligating the right ureter. The UUO model rats were randomly divided into model group, Tanshinone ⅡA group, DAPT group and combination group, 6 in each group. Six rats were recruited as the normal group. The normal group and the model group were administered with CMC-Na by gavage. The medication groups were administered with Tanshinone ⅡA(25 mg/kg), DAPT(12 mg/kg), Tanshinone ⅡA(25 mg/kg) combined with DAPT(12 mg/kg) by gavage respectively. The rats were sacrificed after 2 weeks. The collagen fibers in renal tissue were observed by Masson staining. The expressions of NICD and Hey1 in renal tissues were detected by immunohistochemical. The expressions of Rbp-Jk and ColⅠin renal tissues were determined by Western Blot. Results Compared with the normal group, the Masson score was increased(P<0.05), the protein expressions of NICD, Hey1, Rbp-Jk and ColⅠwere increased in the model group(P<0.05). Compared with the model group, the Masson scores were decreased(P<0.05), the protein expressions of NICD, Hey1, Rbp-Jk and ColⅠwere decreased in the medication groups(P<0.05). Compared with TanshinoneⅡA group and DAPT group, the Masson score was decreased(P<0.05);the protein expressions of NICD, Hey1, Rbp-Jk and ColⅠwere decreased in combination group(P<0.05). Conclusions TanshinoneⅡcombined with DAPT can improve the pathological damage of renal interstitial in UUO model, and alleviate collagen fiber synthesis deposition during renal fibrosis. The mechanism might be correlated to inhibiting the expression of NICD, Rbp-Jk, Hey1 and ColⅠin renal tissue, and affecting the Notch/Jagged signaling pathway.