Huang Bao-Ying, Cao Luo-Yuan, Fu Xian-Guo
The Affiliated Ning-de Municipal Hospital of Fujian University of Traditional Chinese Medicine, Fujian.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 Jul;32(7):965-9.
To observe the expressions of Wnt/beta-catenin and the effects of tanshinone IIA (TII A) on Wnt/beta-catenin signaling pathway in high glucose induced renal tubular epithelial cell transdifferentiation.
Human kidney proximal tubular epithelial cells (HK-2) were divided into three groups, i. e., the normal glucose group, the high glucose group, and the high glucose plus tanshinone IIA group. The expression of beta-catenin was observed using immunocytochemical staining. The protein expression of beta-catenin, E-cadherin, and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot. The mRNA levels of beta-catenin and E-cadherin were detected by RT-PCR.
Compared with the normal glucose group, both the protein and the mRNA expressions of beta-catenin were significantly enhanced (P < 0.01), the expression of E-cadherin significantly decreased (P < 0.01), the expression of beta-catenin increased in the cytoplasm and nucleus in the high glucose group. TIIA at the final concentration of 100 micromol/L significantly reduced the ectopic expression of beta-catenin. At that concentration, the protein and mRNA expressions of beta-catenin in the nucleus significantly decreased, while the protein and mRNA expressions of E-cadherin were up-regulated. Meanwhile, the expression of alpha-SMA obviously decreased.
Wnt/beta-catenin signaling pathway participated in the high glucose induced renal tubular epithelial cell transdifferentiation. TIIA inhibited the transdifferentiation process possibly through down-regulating the activities of Wnt/beta-catenin signaling pathway, thus further playing a role in renal protection.
观察Wnt/β-连环蛋白的表达以及丹参酮IIA(TIIA)对高糖诱导的肾小管上皮细胞转分化中Wnt/β-连环蛋白信号通路的影响。
将人肾近端小管上皮细胞(HK-2)分为三组,即正常葡萄糖组、高糖组和高糖加丹参酮IIA组。采用免疫细胞化学染色观察β-连环蛋白的表达。通过蛋白质印迹法检测β-连环蛋白、E-钙黏蛋白和α-平滑肌肌动蛋白(α-SMA)的蛋白表达。通过逆转录-聚合酶链反应(RT-PCR)检测β-连环蛋白和E-钙黏蛋白的mRNA水平。
与正常葡萄糖组相比,高糖组β-连环蛋白的蛋白和mRNA表达均显著增强(P<0.01),E-钙黏蛋白表达显著降低(P<0.01),β-连环蛋白在细胞质和细胞核中的表达增加。终浓度为100μmol/L的TIIA显著降低了β-连环蛋白的异位表达。在该浓度下,细胞核中β-连环蛋白的蛋白和mRNA表达显著降低,而E-钙黏蛋白的蛋白和mRNA表达上调。同时,α-SMA的表达明显降低。
Wnt/β-连环蛋白信号通路参与了高糖诱导的肾小管上皮细胞转分化。TIIA可能通过下调Wnt/β-连环蛋白信号通路的活性抑制转分化过程,从而进一步发挥肾脏保护作用。