【目的】探讨藁本内酯对脂多糖 （LPS） 诱导小鼠肾足细胞损伤的改善作用及机制。 【方法】（1） 体内实验：将小鼠随机分 为空白对照组，LPS诱导组，藁本内酯低、高剂量组及藁本内酯高剂量+pcDNA-NC （RhoA阴性对照） 组、藁本内酯高剂量+ pcDNA-RhoA （RhoA过表达） 组。除空白对照组，其他各组小鼠采用LPS诱导法构建急性肾损伤 （AKI） 模型。干预结束后，检 测小鼠血清中尿素氮 （BUN） 和肌酐 （Cr） 水平，高碘酸-希夫 （PAS） 染色法观察小鼠肾脏组织的病理变化，Western Blot法检测 肾脏组织中RhoA、Rho相关卷曲螺旋形成蛋白激酶 （ROCK） 蛋白的表达。 （2） 体外实验：将培养的小鼠足细胞MPC5分为空白 对照组，LPS诱导组，藁本内酯低、高剂量组，藁本内酯高剂量+pcDNA-NC组，藁本内酯高剂量+pcDNA-RhoA组，细胞计 数试剂盒8 （CCK-8） 法检测足细胞增殖活性，流式细胞术检测细胞凋亡，鬼笔环肽染色法分析细胞骨架，Western Blot法检测 细胞RhoA、ROCK蛋白的表达。 【结果】与空白对照组比较，LPS诱导组小鼠血清中BUN和Cr的水平、肾小管损伤评分、 RhoA和ROCK蛋白表达水平升高 （P＜0.05） ；与LPS诱导组比较，藁本内酯低、高剂量组小鼠血清中BUN和Cr水平、肾小管 损伤评分、RhoA和ROCK表达水平降低 （P＜0.05） 。与空白对照组比较，LPS诱导组MPC5细胞增殖活性、F-肌动蛋白荧光 强度、RhoA和ROCK的表达降低 （P＜0.05） ，凋亡率升高 （P＜0.05） ；与LPS诱导组比较，藁本内酯低、高剂量组MPC5细胞 增殖活性、F-肌动蛋白荧光强度、RhoA和ROCK的表达升高 （P＜0.05） ，凋亡率降低 （P＜0.05） 。利用过表达RhoA进行回补 实验发现，RhoA过表达逆转了藁本内酯对LPS诱导小鼠肾组织和足细胞损伤的缓解作用 （P＜0.01） 。 【结论】藁本内酯可能通 过抑制RhoA/ROCK信号通路改善LPS诱导小鼠肾足细胞损伤。

Objective To investigate the improvement effect and mechanism of ligustilide on lipopolysaccharide （LPS） -induced renal podocyte injury in mice. Methods （1） In vivo experiment：the mice were randomly divided into blank control group，LPS induction group，ligustilide low- and high- dose groups，ligustilide high-dose + pcDNA-NC （RhoA negative control） group，ligustilide high-dose + pcDNA-RhoA （RhoA overexpression） group. Except for the blank control group，mice in other groups were subjected to LPS-induced acute kidney injury （AKI） model. After the intervention，the levels of blood urea nitrogen （BUN） and creatinine （Cr） in the serum of mice were detected，the pathological changes of kidney tissue in mice were observed by periodic acid-schiff （PAS） staining，and the protein expressions of RhoA and Rho-associated coiled-coil forming protein kinase （ROCK） in kidney tissue were detected by Western Blot. （2） In vitro experiments：the cultured mouse podocyte MPC5 was divided into blank control group，LPS induction group，ligustilide low- and high- dose groups， ligustilide high-dose + pcDNA-NC group，ligustilide high-dose + pcDNA-RhoA group. Cell count kit 8 （CCK-8） method was used to detect the proliferation activity of podocytes，flow cytometry was used to detect apoptosis， phalloidin staining method was used to analyze cytoskeleton，and Western Blot was used to detect the expressions of RhoA and ROCK proteins in the cells. Results Compared with the blank control group，the levels of BUN and Cr in serum，renal tissue injury score，and the protein expressions of RhoA and ROCK in the LPS-induced group were increased （P＜0.05） . Compared with the LPS-induced group，the levels of BUN and Cr in serum，renal tissue injury score，and the expression levels of RhoA and ROCK in the low- and high-dose ligustilide groups were decreased （P＜0.05） . Compared with the blank control group， the cell proliferation activity， F-actin fluorescence intensity，RhoA and ROCK expressions in the LPS-induced group were decreased（P＜0.05） ，and the apoptosis rate was increased （P＜0.05） . Compared with the LPS-induced group， the cell proliferation activity， F-actin fluorescence intensity， RhoA and ROCK expressions were increased（P＜0.05）， and the apoptosis rate was decreased in the low- and high-dose ligustilide groups（P＜0.05） . Overexpression of RhoA reversed the alleviation effect of ligustilide on LPS-induced renal tissue and podocyte injury in mice（P＜0.01） . Conclusion Ligustilide may improve LPS-induced renal podocyte injury in mice by inhibiting RhoA/ROCK signaling pathway.

R285.5

江苏省高等学校自然科学研究面上项目 （编号：21KJB350024）