【目的】观察丹酚酸B对肾纤维化小鼠的改善作用及机制。【方法】将36只BALB/c小鼠随机分为假手术组，模型组，丹酚酸B低、中、高剂量组及VX765[半胱氨酸天冬氨酸蛋白酶1（caspase-1）抑制剂]组，每组6只。丹酚酸B低、中、高剂量组分别以相应剂量50、100、200mg•kg-1•d-1丹酚酸B生理盐水混悬液灌胃，VX765组以50mg•kg-1•d-1 VX765生理盐水混悬液灌胃，假手术组和模型组灌胃等体积生理盐水，每日1次。7d后，除假手术组外，其他各组小鼠采用单侧肾脏缺血再灌注损伤联合对侧肾切除术建立急性肾损伤后肾纤维化模型。造模后各组继续灌胃相应体积药物21d。给药结束后，采用苏木素-伊红（HE）染色及马松（Masson）染色观察肾组织病理学变化，采用肌氨酸氧化酶法检测血清肌酐（SCr），采用脲酶法检测尿素氮（BUN），酶联免疫吸附分析（ELISA）检测血清中性粒细胞明胶酶相关脂质运载蛋白（NGAL）水平，蛋白质免疫印迹（Western Blot）法检测肾组织纤维化蛋白α-平滑肌肌动蛋白（α-SMA）、波形蛋白（vimentin）、转化生长因子β（TGF-β）及焦亡相关蛋白裂解型半胱氨酸天冬氨酸蛋白水解酶1（cl-caspase-1）caspase-1前体（pro-caspase-1）、NOD样受体热蛋白结构域相关蛋白3（NLRP3）、裂解型白细胞介素1β（cl-IL-1β）、白细胞介素1β（IL-1β）、消皮素D-N端片段（GSDMD-N）、消皮素D（GSDMD）表达。【结果】与假手术组比较，模型组小鼠血清SCr、BUN、NGAL水平升高（P＜0.05或P＜0.01），HE及Masson染色显示肾间质有炎性细胞浸润及大量胶原沉积，纤维化相关蛋白α-SMA、vimentin、TGF-β表达水平升高（P＜0.01），焦亡相关蛋白cl-caspase-1/pro-caspase-1、NLRP3、cl-IL-1β/IL-1β、GSDMD-N/GSDMD表达水平亦升高（P＜0.05或P＜0.01）；与模型组比较，丹酚酸B高剂量组血清SCr、BUN、NGAL水平显著降低（P＜0.05或P＜0.01），肾间质炎性细胞浸润及胶原沉积减轻，纤维化蛋白α-SMA、vimentin、TGF-β及焦亡相关蛋白cl-caspase-1/pro-caspase-1、NLRP3、cl-IL-1β/IL-1β、GSDMD-N/GSDMD表达水平降低（P＜0.05或P＜0.01）。丹酚酸B高剂量组上述各指标与VX765组比较，差异均无统计学意义（P＞0.05）。【结论】丹酚酸B可能通过抑制NLRP3/caspase-1/GSDMD通路介导的细胞焦亡，减轻小鼠肾纤维化。

Objective To observe the improvement effect and mechanism of salvianolic acid B on renal fibrosis mice. Methods Thirty-six BALB/c mice were randomly divided into sham operation group，model group，salvianolic acid B low-，medium- and high- dose groups and VX765 （caspase-1 inhibitor）group，with 6 mice in each group. Salvianolic acid B low-，medium- and high- dose groups were intragastrically administered with corresponding doses of 50，100，200 mg•kg-1•d-1 salvianolic acid B normal saline suspension，respectively，VX765 group was intragastrically administered with 50 mg•kg-1•d-1 VX765 normal saline suspension，and the sham operation group and the model group were intragastrically administered with the same volume of normal saline，once a day. After seven days，except for the sham operation group，the mice in other groups were treated with unilateral renal ischemia-reperfusion injury combined with contralateral nephrectomy to establish an acute kidney injury-induced renal fibrosis model. After modeling，each group continued to be administered with the corresponding volume of drugs for 21 days. After the administration，the pathological changes of renal tissue were observed by hematoxylin-eosin（HE）staining and Masson staining. The serum creatinine（SCr）was detected by sarcosine oxidase method. The urea nitrogen（BUN）was detected by urease method. The level of serum neutrophil gelatinase-associated lipocalin（NGAL）was detected by enzyme-linked immunosorbent assay（ELISA）. The expression of fibrosis-related proteins fibrosis-related proteins α-smooth muscle actin（α-SMA），vimentin and transforming growth factor-β（TGF-β）and pyroptosis-related proteins cleaved cysteine aspartate proteolytic enzyme 1（cl-caspase-1），pro-caspase-1（pro-caspase-1），NOD-like receptor hot protein domain-related protein 3（NLRP3），cleaved interleukin-1β（cl-IL-1β），interleukin-1β（IL-1β），GSDMD-N，GSDMD in renal tissue was detected by Western Blot. Results Compared with the sham operation group，the serum levels of SCr，BUN and NGAL in the model group were increased （P＜0.05 or P＜0.01）. HE and Masson staining showed inflammatory cell infiltration and a large amount of collagen deposition in the renal interstitium，and the expression levels of fibrosis-related proteins α-SMA，vimentin and TGF-βwere increased（P＜0.01） . The expression levels of pyroptosis-related proteins cl-caspase-1/pro-caspase-1，NLRP3，cl-IL-1β/IL-1β，GSDMD-N/GSDMD were also increased（P＜0.05 or P＜0.01）. Compared with the model group，the levels of serum SCr，BUN and NGAL in the high-dose salvianolic acid B group were significantly decreased （P＜0.05 or P＜0.01），the infiltration of renal interstitial inflammatory cells and collagen deposition were reduced，and the expression levels of fibrosis proteins α-SMA，vimentin，TGF-βand pyroptosis-related proteins cl-caspase-1/pro-caspase-1，NLRP3，cl-IL-1β/IL-1β，GSDMD-N/GSDMD were decreased（P＜0.05 or P＜0.01）. There was no significant difference in the above indexes between the high-dose salvianolic acid B group and the VX765 group（P＞0.05）. Conclusion Salvianolic acid B may alleviate renal fibrosis in mice by inhibiting pyroptosis mediated by NLRP3/caspase-1/GSDMD pathway.

R285.5

国家自然科学基金资助项目 （编号：81803824，82174061）