目的 研究抑瘤湯對(duì)上皮性卵巢癌中NOD樣受體熱蛋白結(jié)構(gòu)域相關(guān)蛋白3（NLRP3）炎癥小體的作用。方法 通過(guò)免疫組化法比較人體的良性腫物和卵巢癌組織中NLRP3和白細(xì)胞介素（IL）-1β的表達(dá)水平；體外培養(yǎng)人上皮性卵巢癌細(xì)胞系A(chǔ)2780，分成對(duì)照組、化療（給予DMSO溶解的紫杉醇100nmol·L^-1）組、化療＋抑瘤湯（1、10、100mg·L^-1）組，對(duì)照組加入等量DMSO，干預(yù)12、24、48、72h后，CCK-8法檢測(cè)細(xì)胞存活率，選取合適的抑瘤湯濃度及干預(yù)時(shí)間進(jìn)行后續(xù)實(shí)驗(yàn)；Western blotting法檢測(cè)NLRP3的蛋白表達(dá)；實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測(cè)NLRP3、半胱氨酸蛋白酶-1（Caspase-1）、凋亡相關(guān)斑點(diǎn)樣蛋白（ASC）和IL-1β的mRNA水平；ELISA法檢測(cè)細(xì)胞上清液中NLRP3、Caspase-1、ASC和IL-1β蛋白水平。結(jié)果 與卵巢良性腫物組相比，卵巢癌患者的卵巢組織中NLRP3及IL-1β蛋白表達(dá)顯著上調(diào)（P＜0.05）；干預(yù)24、48、72h后，與對(duì)照組相比，化療組的細(xì)胞存活率顯著降低（P＜0.05、0.001）；與化療組相比，化療＋100mg·L^-1抑瘤湯組的細(xì)胞存活率顯著降低（P＜0.01、0.001），選取100mg·L^-1抑瘤湯干預(yù)48h進(jìn)行后續(xù)研究。與對(duì)照組相比，化療組的細(xì)胞中NLRP3蛋白表達(dá)水平降低，但無(wú)統(tǒng)計(jì)學(xué)意義；與對(duì)照組及化療組相比，化療＋抑瘤湯組的細(xì)胞中NLRP3蛋白表達(dá)水平顯著降低（P＜0.05、0.001）。與對(duì)照組相比，化療組和化療＋抑瘤湯組的NLRP3 mRNA和上清中蛋白水平顯著降低（P＜0.05、0.001）；與化療組相比，化療＋抑瘤湯組的NLRP3mRNA和上清中蛋白水平顯著降低（P＜0.05）。與對(duì)照組相比，化療組的Caspase-1、ASC、IL-1β的mRNA水平降低，但無(wú)統(tǒng)計(jì)學(xué)意義；化療＋抑瘤湯組的Caspase-1、ASC、IL-1β mRNA水平顯著降低（P＜0.05）。與對(duì)照組相比，化療組和化療＋抑瘤湯組細(xì)胞上清中的Caspase-1、ASC、IL-1β蛋白水平顯著降低（P＜0.01、0.001）；與化療組相比，化療＋抑瘤湯組的Caspase-1、ASC、IL-1β蛋白進(jìn)一步降低，其中Caspase-1、ASC差異顯著（P＜0.05、0.001）。結(jié)論 NLRP3炎癥小體在卵巢癌中高表達(dá)，抑瘤湯可抑制卵巢癌細(xì)胞NLRP3炎癥小體的表達(dá)。

Objective To study the effect of Yiliu Decoction on Nod-like receptor pyrin domain 3 (NLRP3) inflammasome in epithelial ovarian cancer.Methods Expression of NLRP3 and interleukin (IL) -1β in benign tumor and ovarian cancer patients were compared by immunohistochemistry. Human epithelial ovarian cancer cell line A2780 was cultured in vitro and divided into control group, chemotherapy (DMSO-dissolved paclitaxel 100 nmol·L^-1) group, chemotherapy + Yiliu Decoction (1, 10 and 100 mg·L^-1) group. The control group was treated with DMSO. After 12, 24, 48 and 72 h, survival rate of cells was detected by CCK-8 method, and the appropriate concentration and intervention time of antitumor soup were selected for follow-up experiments. The protein expression of NLRP3 was detected by Western blotting. The mRNA levels of NLRP3, Caspase-1, apoptosis-related spot-like protein (ASC) and IL-1β were detected by real-time quantitative fluorescence PCR (qRT-PCR). The levels of NLRP3, Caspase-1, ASC and IL-1β in the supernatant were determined by ELISA.Results Compared with benign ovarian tumor group, NLRP3 and IL-1β protein expressions in ovarian tissue of ovarian cancer patients were significantly up-regulated (P <0.05). After 24, 48, 72 h of intervention, the survival rate of cells in chemotherapy group was significantly decreased compared with control group (P <0.05, 0.001). Compared with the chemotherapy group, the survival rate of cells in the chemotherapy +100 mg·L^-1 Yiliu Decoction group was significantly decreased (P <0.01, 0.001). The 100 mg·L^-1 Yiliu Decoction was selected to intervene for 48 h for follow-up study. Compared with the control group, the expression level of NLRP3 protein in the chemotherapy group was decreased, but there was no statistical significance. Compared with the control group and the chemotherapy group, the NLRP3 protein expression level in the chemotherapy + Yiliu Decoction group was significantly decreased (P <0.05, 0.001). Compared with control group, NLRP3 mRNA and protein levels in supernatant of chemotherapy group and chemotherapy + Yiliu Decoction group were significantly decreased (P <0.05, 0.001). Compared with chemotherapy group, NLRP3 mRNA and protein levels in supernatant of chemotherapy + Yiliu Decoction group were significantly decreased (P <0.05). Compared with control group, mRNA levels of Caspase-1, ASC and IL-1β in the chemotherapy group were decreased, but there was no statistical significance. The mRNA levels of Caspase-1, ASC and IL-1β in chemotherapy + Yiliu Decoction group were significantly decreased (P <0.05). Compared with control group, the protein levels of Caspase-1, ASC and IL-1β in cell supernatant of chemotherapy group and chemotherapy + antitumor decoction group were significantly decreased (P <0.01, 0.001). Compared with the chemotherapy group, the proteins of Caspase-1, ASC and IL-1β in the chemotherapy + antitumor decoction group were further decreased, and the differences of Caspase-1 and ASC were significant (P <0.05, 0.001).Conclusion NLRP3 inflammasome is highly expressed in ovarian cancer. Yiliu Decoction inhibits the expression of NLRP3 inflammasome in ovarian cells.

R285.5

福建省中青年教師教育科研項(xiàng)目(JAT190249);福建省自然科學(xué)基金面上項(xiàng)目(2020J011041);正大天晴醫(yī)學(xué)項(xiàng)目研究(201511669);福建省衛(wèi)生計(jì)生科研人才培養(yǎng)項(xiàng)目-醫(yī)學(xué)創(chuàng)新課題(2018-CX-44);福建省自然科學(xué)基金青創(chuàng)項(xiàng)目(2020J05061)