Mao Xin, Yao Rong-Mei, Bao Yan-Yan, Sun Jing, Cui Xiao-Lan, Zhang Hai-Jiang, Shi Yu-Jing
Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China School of Pharmaceutical Sciences,Southern Medical University Guangzhou 510515,China.
Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China Institute of Chinese Medicine,Tianjin University of Traditional Chinese Medicine Tianjin 300193,China.
Zhongguo Zhong Yao Za Zhi. 2021 Jul;46(13):3388-3393. doi: 10.19540/j.cnki.cjcmm.20210308.401.
To study the mechanism of polysaccharides from seeds of Vaccaria segetalis( PSV) in the treatment of bacterial cystitis through the NLRP3 inflammasome pathway. The rat model of urinary tract infection was used and treated with PSV,and the urine and bladders were collected. The level of interleukin-10( IL-10) in rat urine was detected by enzyme linked immunosorbent assay( ELISA). Western blot and immunofluorescence staining were used to detect the expressions of sonic hedgehog( SHH) and NLRP3 inflammasome [NOD-like receptor thermoprotein domain 3( NLRP3),apoptosis associated speck like protein( ASC) and pro-caspase-1]. The expression of Toll-like receptor pathway was detected by RT-PCR. The death of 5637 cells induced by uropathogenic Escherichia coli( UPEC) and lactate dehydrogenase( LDH) release were evaluated using live/dead staining. The results showed that in the rat bladder,the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors were significantly up-regulated,and NLRP3 inflammasomes were significantly activated by UPEC infection. The administration with PSV could significantly increase the concentration of IL-10 in urine,inhibit the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors in bladder,and inhibit the activation of NLRP3 inflammasomes. A large number of 5637 cells were dead after UPEC infection and caused LDH production. PSV could significantly inhibit the death of 5637 cells and the release of LDH. In conclusion,PSV could inhibit the expression and activation of NLRP3 inflammasomes by inhibiting the Toll-like receptor pathway,thereby mitigating the bladder injury.
研究王不留行籽多糖(PSV)通过NLRP3炎性小体途径治疗细菌性膀胱炎的机制。采用大鼠尿路感染模型并给予PSV治疗,收集尿液和膀胱组织。采用酶联免疫吸附测定(ELISA)检测大鼠尿液中白细胞介素10(IL-10)水平。采用蛋白质免疫印迹法和免疫荧光染色法检测音猬因子(SHH)和NLRP3炎性小体[核苷酸结合寡聚化结构域样受体热蛋白结构域3(NLRP3)、凋亡相关斑点样蛋白(ASC)和前半胱天冬酶-1]的表达。采用逆转录-聚合酶链反应(RT-PCR)检测Toll样受体途径的表达。使用活/死染色评估尿路致病性大肠杆菌(UPEC)诱导的5637细胞死亡和乳酸脱氢酶(LDH)释放。结果显示,在大鼠膀胱中,SHH、NLRP3炎性小体和Toll样受体的表达显著上调,UPEC感染显著激活NLRP3炎性小体。给予PSV可显著提高尿液中IL-10浓度,抑制膀胱中SHH、NLRP3炎性小体和Toll样受体的表达,并抑制NLRP3炎性小体的激活。UPEC感染后大量5637细胞死亡并导致LDH产生。PSV可显著抑制5637细胞死亡和LDH释放。综上所述,PSV可通过抑制Toll样受体途径抑制NLRP3炎性小体的表达和激活,从而减轻膀胱损伤。