对比剂肾病大鼠肾脏bax及bcl-2表达及羟苯磺酸钙干预研究

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摘要目的研究对比剂肾病（CIN）大鼠肾脏bax及bcl-2的表达情况及羟苯磺酸钙对干预效果。方法将84只大鼠按照随机数字表法分为假手术组（A组）、模型组（B组）和羟苯磺酸钙干预组（C组），每组28只。A组给予生理盐水，B组将INDO、L-NAME、760 g/L泛影葡胺进行注射，C组在B组基础上给予羟苯磺酸钙灌胃。造模后12、24、48、72 h测定血清尿素氮（BUN）和肌酐（Scr），HE染色观察肾病理改变，TUNEL检测肾脏细胞凋亡，免疫组化和Western blot检测肾脏bax及bcl-2的表达。结果 B组和C组造模后12、24、48、72 h血清BUN和Scr水平显著高于A组同期，C组造模后12、24、48、72 h血清BUN和Scr水平显著低于B组同期（P < 0.05）；与造模后12 h比较，B组和C组造模后24、48、72 h血清BUN和Scr水平均显著升高（P < 0.05）；与造模后24 h比较，B组造模后48 h血清Scr水平显著升高，C组造模后48 h血清BUN和Scr水平均显著升高，C组造模后72 h血清Scr水平显著降低（P < 0.05）；与造模后48 h比较，B组和C组造模后72 h血清BUN和Scr水平均显著降低（P < 0.05）。A组病理改变不显著；B组造模后12 h出现广泛肾小管上皮细胞空泡样变性、部分肾小管坏死，肾间质纤维化；C组肾小管间质病变显著改善。A组大鼠肾细胞无明显凋亡；B组大鼠肾细胞凋亡指数显著高于A组，C组肾细胞凋亡指数显著低于B组，但高于A组（P < 0.05）。B组大鼠bax表达水平显著高于A组，C组bax表达水平显著低于B组，但高于A组；B组大鼠bcl-2表达水平显著低于A组，C组bcl-2表达水平显著高于B组，但低于A组（P < 0.05）。B组大鼠bax蛋白相对表达量显著高于A组，C组bax蛋白相对表达量显著低于B组，但高于A组；B组大鼠bcl-2蛋白相对表达量显著低于A组，C组bcl-2蛋白相对表达量显著高于B组，但低于A组（P < 0.05）。结论细胞凋亡可能参与大鼠CIN的发生、发展，且羟苯磺酸钙可能通过减少bax的表达及增加bcl-2表达减轻肾脏细胞凋亡，发挥保护作用。

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	李伟1
	2 于芬芬3 季文萱3 何晓艳1 潘配强1
	3 孙艳4 黄俊彦3

关键词 ：对比剂肾病, 细胞凋亡, bax, bcl-2, 羟苯磺酸钙

Abstract：Objective To investigate the expression of bax and bcl-2 in kidney of rats with contrast-induced nephropathy (CIN) and the effects of Calcium Dobesilate. Methods Eighty-four rats were divided into sham operation group (group A), model group (group B) and Calcium Dobesilate intervention group (group C) according to random number table, 28 rats in each group. Group A was given saline, group B was injected with INDO, L-NAME and 760 g/L Diatrizoate, while group C was given Calcium Dobesilate for intragastric administration on the basis of group B. After 12, 24, 48, 72 h model establishment, blood urea nitrogen (BUN) and creatinine (Scr) were measured. Nephropathy was observed by HE staining. Renal cell apoptosis was detected by TUNEL. The expression of bax and bcl-2 were detected by immunohistochemistry and Western blot. Results The levels of BUN and Scr in group B and group C were significantly higher than those in group A at 12, 24, 48, 72 h after the establishment of model, while those in group C were significantly lower than those in group B at 12, 24, 48, 72 h after the establishment of model (P < 0.05). Compared with 12 h after modeling, serum BUN and Scr levels in group B and group C were significantly higher at 24, 48, 72 h after modeling (P < 0.05). Compared with 24 h after model establishment, serum Scr level in group B increased significantly at 48 h after model establishment, serum BUN and Scr levels in group C increased significantly at 48 h after model establishment, and serum Scr level in group C decreased significantly at 72 h after model establishment (P < 0.05). Compared with 48 h after modeling, serum BUN and Scr levels in group B and group C were significantly lower at 72 h after modeling (P < 0.05). In group A, the pathological changes were not significant; in group B, extensive tubular epithelial cell vacuolation, partial tubular necrosis and renal interstitial fibrosis appeared 12 h after modeling; in group C, the renal tubular interstitial lesions were significantly improved. There was no significant apoptosis in group A; the apoptosis index in group B was significantly higher than that in group A; the apoptosis index in group C was significantly lower than that in group B, but higher than that in group A (P < 0.05). The expression level of bax in group B was significantly higher than that in group A, bax in group C was significantly lower than that in group B, but higher than that in group A; the expression level of bcl-2 in group B was significantly lower than that in group A, and the expression level of bcl-2 in group C was significantly higher than that in group B, but lower than that in group A (P < 0.05). The relative expression of bax protein in group B was significantly higher than that in group A, and the relative expression of bax protein in group C was significantly lower than that in group B, but higher than that in group A; the relative expression of bcl-2 protein in group B was significantly lower than that in group A, and the relative expression of bcl-2 protein in group C was significantly higher than that in group B, but lower than that in group A (P < 0.05). Conclusion Apoptosis has close relationship with CIN, and Calcium Dobesilate may play a protective role by reducing the expression of bax and increasing the expression of bcl-2.

Key words： Contrast-induced nephropathy Apoptosis bax bcl-2 Calcium Dobesilate

基金资助:山东省医药卫生科技发展计划项目（2018WS359）；
山东省青岛市民生科技计划项目（16-6-2-19-nsh）。

通讯作者: 黄俊彦（1963-），男，硕士，主任医师，硕士研究生导师；研究方向：急性肾损伤。

作者简介: 李伟（1982.3-），女，青岛大学医学部2015级肾内科在读硕士研究生；研究方向：急性肾损伤。

引用本文:

李伟1，2 于芬芬3 季文萱3 何晓艳1 潘配强1，3 孙艳4 黄俊彦3. 对比剂肾病大鼠肾脏bax及bcl-2表达及羟苯磺酸钙干预研究[J]. 中国医药导报, 2019, 16(32): 9-13.
LI Wei1,2 YU Fenfen3 JI Wenxuan3 HE Xiaoyan1 PAN Peiqiang1,3 SUN Yan4 Huang Junyan3. Study on the expression of bax and bcl-2 in kidney of rats with contrast-induced nephropathy and the intervention of Calcium Dobesilate. 中国医药导报, 2019, 16(32): 9-13.

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http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2019/V16/I32/9

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