川芎嗪对肾间质纤维化中TGF-β1/miR-29表达的影响
| 作者: |
1,2徐梅,
1,2孙升云
1 暨南大学医学院中医学院，广州 510632 2 暨南大学第一附属医院广州华侨医院中医科，广州 510632 |
| 通讯: |
孙升云
Email: gzssy@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2017.05.002 |
| 基金: | 广东省中医药局科研项目, 20171075 广东省科技计划项目, 2014A020221015 |
摘要
目的：观察川芎嗪(tetramethylpyrazine，TMP)对单侧输尿管结扎(unilateral ureteral obstruction，UUO)大鼠模型肾间质纤维化(renal interstitial fibrosis，RIF)、间质损伤及TGF-β1/miR-29表达的影响。方法：50只雄性Wistar大鼠随机分为5组：正常对照组(Ctrl组)、假手术组(Sham组)、模型组(UUO组)、缬沙坦组(VAN组)和TMP组各10只。经相应处理后，通过HE，Masson染色观察肾组织的病理学表现；同时采用SP免疫组织化学法检测各组病变组织中TGF-β1的表达情况；最后采用RT-PCR技术比较各组TGF-β1，miR-29基因的表达。结果：HE染色结果显示Ctrl组、Sham组、UUO组、VAN组及TMP组的肾间质损伤评分分别为0.27±0.10，0.30±0.12，8.79±1.03，6.23±0.81及4.57±0.74。Masson染色胶原半定量分析结果显示Ctrl组、Sham组、UUO组、VAN组及TMP组的评分分别为0.21±0.03，0.23±0.06，2.49±0.33，1.63±0.07及1.32±0.04。SP免疫组织化学染色显示模型组TGF-β1的蛋白表达高于对照组；而缬沙坦和TMP可部分逆转TGF-β1蛋白的表达，且TMP的效果优于缬沙坦。RT-PCR结果显示模型组的TGF-β1 mRNA表达增高、miR-29 mRNA表达降低。TMP组和缬沙坦组TGF-β1 mRNA表达明显低于模型组，而miR-29 mRNA表达则明显高于模型组；且TMP组TGF-β1 mRNA表达高于缬沙坦组。结论：缬沙坦和TMP可明显减轻UUO致RIF中肾组织学损伤，且TMP的疗效优于缬沙坦。此外TMP还可抑制TGF-β1而促进miR-29的表达。
关键词：
单侧输尿管结扎术
肾间质纤维化
川芎嗪
TGF-β1
miR-29
Effect of tetramethylpyrazine on TGF-β1/miR-29 expression in renal interstitial fibrosis
CorrespondingAuthor: SUN Shengyun Email: gzssy@163.com
DOI: 10.3978/j.issn.2095-6959.2017.05.002
Abstract
Objective: To observe the effect of tetramethylpyrazine (TMP) on renal interstitial fibrosis, interstitial injury and TGF-β1/miR-29 expression in UUO model rats. Methods: Fifty male Wistar rats were randomly divided into five groups: normal control group (Ctrl), sham operation group (Sham), model group (UUO), valsartan group (VAN) and TMP group (TMP) with 10 in each group. After corresponding treatment, all rats were sacrificed and the left renal were used to evaluate pathology changes of rats’ kidney tissue with HE and Masson staining; while using SP immunohistochemical method to detect the lesions in the expression of TGF-β1; and finally compare the gene expression of TGF-β1 and miR-29 in each group of by RT-PCR assay. Results: HE staining results showed that the interstitial injury scores of Ctrl, Sham, UUO, VAN and TMP group were 0.27±0.10, 0.30±0.12, 8.79±1.03, 6.23±0.81 and 4.57±0.74, respectively. The results of Masson staining showed that scores of Ctrl, Sham, UUO, VAN and TMP group were 0.21±0.03, 0.23±0.06, 2.49±0.33, 1.63±0.07 and 1.32±0.04, respectively. SP immunohistochemical staining showed that compared with the control group, model group has a higher expression of TGF-β1, while treatment with valsartan and TMP could reverse the up-regulation of TGF-β1 and TMP is more effective than valsartan. RT-PCR results showed that TGF-β1 mRNA expression was increased, while miR-29 mRNA expression decreased in model group. VAN group and TMP group have a significantly lower expression of TGF-β1 mRNA level compared to the model group, while miR-29 mRNA expression was significantly higher in both VAN group and TMP group; and TGF-β1 mRNA expression in the TMP group was higher than that in VAN group. Conclusion: Valsartan and TMP could significantly reduce the renal histological damage in renal interstitial fibrosis and TMP is more effective than valsartan. Furthermore, TMP could inhibit TGF-β1 gene expression, while promoting the miR-29 expression.