三七总皂苷通过下调lncRNA-ANRIL抑制4型心肾综合征中NLRP3炎性小体介导的细胞焦亡。

Panax notoginseng saponins inhibits NLRP3 inflammasome-mediated pyroptosis by downregulating lncRNA-ANRIL in cardiorenal syndrome type 4.

作者信息

Xu Ying, Cao Luxi, Zou Wenli, Yu Rizhen, Shen Wei

机构信息

Urology & Nephrology Center, Department of Urology, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, 310014, Zhejiang, China.

Urology & Nephrology Center, Department of Nephrology, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, No. 158, Shangtang Road, Hangzhou, 310014, Zhejiang, China.

出版信息

Chin Med. 2023 May 8;18(1):50. doi: 10.1186/s13020-023-00756-2.

OBJECTIVE

Cardiorenal syndrome type 4 (CRS4) is a complication of chronic kidney disease. Panax notoginseng saponins (PNS) have been confirmed to be efficient in cardiovascular diseases. Our study aimed to explore the therapeutic role and mechanism of PNS in CRS4.

METHODS

CRS4 model rats and hypoxia-induced cardiomyocytes were treated with PNS, with and without pyroptosis inhibitor VX765 and ANRIL overexpression plasmids. Cardiac function and cardiorenal function biomarkers levels were measured by echocardiography and ELISA, respectively. Cardiac fibrosis was detected by Masson staining. Cell viability was determined by cell counting kit-8 and flow cytometry. Expression of fibrosis-related genes (COL-I, COL-III, TGF-β, α-SMA) and ANRIL was examined using RT-qPCR. Pyroptosis-related protein levels of NLRP3, ASC, IL-1β, TGF-β1, GSDMD-N, and caspase-1 were measured by western blotting or immunofluorescence staining.

RESULTS

PNS improved cardiac function, and inhibited cardiac fibrosis and pyroptosis in a dose-dependent manner in model rats and injured H9c2 cells (p < 0.01). The expression of fibrosis-related genes (COL-I, COL-III, TGF-β, α-SMA) and pyroptosis-related proteins (NLRP3, ASC, IL-1β, TGF-β1, GSDMD-N, and caspase-1) was inhibited by PNS in injured cardiac tissues and cells (p < 0.01). Additionally, ANRIL was upregulated in model rats and injured cells, but PNS reduced its expression in a dose-dependent manner (p < 0.05). Additionally, the inhibitory effect of PNS on pyroptosis in injured H9c2 cells was enhanced by VX765 and reversed by ANRIL overexpression, respectively (p < 0.05).

CONCLUSION

PNS inhibits pyroptosis by downregulating lncRNA-ANRIL in CRS4.

目的

心肾综合征4型（CRS4）是慢性肾脏病的一种并发症。三七总皂苷（PNS）已被证实对心血管疾病有效。本研究旨在探讨PNS在CRS4中的治疗作用及机制。

方法

用PNS处理CRS4模型大鼠和缺氧诱导的心肌细胞，同时分别使用和不使用焦亡抑制剂VX765以及ANRIL过表达质粒。分别通过超声心动图和酶联免疫吸附测定法（ELISA）测量心功能和心肾功能生物标志物水平。通过Masson染色检测心肌纤维化。使用细胞计数试剂盒-8和流式细胞术测定细胞活力。采用逆转录定量聚合酶链反应（RT-qPCR）检测纤维化相关基因（I型胶原、III型胶原、转化生长因子-β、α-平滑肌肌动蛋白）和ANRIL的表达。通过蛋白质免疫印迹法或免疫荧光染色测定NLRP3、凋亡相关斑点样蛋白（ASC）、白细胞介素-1β、转化生长因子-β1、Gasdermin D-N端片段（GSDMD-N）和半胱天冬酶-1等焦亡相关蛋白水平。

结果

PNS以剂量依赖性方式改善模型大鼠的心功能，并抑制心肌纤维化和焦亡，在损伤的H9c2细胞中也有同样效果（p < 0.01）。PNS抑制损伤心脏组织和细胞中纤维化相关基因（I型胶原、III型胶原、转化生长因子-β、α-平滑肌肌动蛋白）和焦亡相关蛋白（NLRP3、ASC、白细胞介素-1β、转化生长因子-β1、GSDMD-N和半胱天冬酶-1）的表达（p < 0.01）。此外，模型大鼠和损伤细胞中ANRIL上调，但PNS以剂量依赖性方式降低其表达（p < 0.05）。此外，VX765增强了PNS对损伤H9c2细胞焦亡的抑制作用，而ANRIL过表达则逆转了这种作用（p < 0.05）。