Tao Hong, Tang Rang-Rang, Su Qing, Huang Li, Li Li-Li, Wu De-Ling, Huang Qi
School of Pharmacy, Anhui University of Chinese Medicine Hefei 230012, China.
School of Pharmacy, Bozhou Vocational and Technical College Bozhou 236000, China.
Zhongguo Zhong Yao Za Zhi. 2025 Jun;50(11):3024-3034. doi: 10.19540/j.cnki.cjcmm.20250306.301.
This paper investigated the inhibitory effect of carbonized Scutellariae Radix(Cb-SR) on pyroptosis in endometrial epithelial cells of mice with endometritis and its correlation with the IKBKE/NLRP3 signaling axis. Mice model of endometritis was established by using an intrauterine injection of 10 μL polysaccharides(LPS, 5 mg·mL(-1)), and the mice were randomly divided into model group(LPS), low-dose group of Cb-SR(L-Cb-SR, 0.55 g·kg(-1)), medium-dose group of Cb-SR(M-Cb-SR, 1.10 g·kg(-1)), high-dose group of Cb-SR(H-Cb-SR, 2.20 g·kg(-1)), crude Scutellariae Radix group(Cr-SR, 1.63 g·kg(-1)), and Fuke Qianjin Capsule group(FQC, 0.30 g·kg(-1)), with 10 mice in each group. Ten healthy female mice were selected and injected with PBS of equal volume into the bilateral uterus, and they were set as the sham group. The mice in the drug treatment groups were given the corresponding doses of Cb-SR, Cr-SR, FQC, or physiological saline of equal volume by gavage twice a day for seven days. Thirty minutes after the last administration, each mouse was euthanized by cervical dislocation. Hematoxylin-eosin(HE) staining and transmission electron microscopy were applied to observe the histopathological morphology of the uterine tissue. Immunohistochemistry was used to detect the expression of CD38 and CD138. Myeloperoxidase(MPO) values in neutrophils were measured by the kit; Enzyme-linked immunosorbent assay(ELISA) was used to measure the secretion of interleukin-18(IL-18), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α). Immunofluorescence and Western blot were used to analyze the expression of the proteins related to the IKBKE/NLRP3 signaling axis. Mouse endometrial epithelial cells(MEECs) were separated and purified from the uterine tissue of pregnant female mice through in vitro experiments and injured by LPS for 24 h, and then they were cultured with Cb-SR-containing serum. The anti-endometritis effect of Cb-SR was investigated by CCK-8 assay, scanning electron microscopy, and Western blot. The results showed that Cb-SR significantly reduced MPO values, attenuated uterine tissue damage, inhibited the expression of CD38 and CD138, decreased the levels of IL-1β, IL-18, and TNF-α, and inhibited the expression of proteins associated with IKBKE/NLRP3 signaling axis in mice with endometritis. In addition, Cb-SR-containing serum reduced swelling of MEECs organelles induced by LPS, decreased the expression of inflammatory factors, and suppressed the expression of IKBKE/NLRP3 signaling axis-related proteins. These results suggest that Cb-SR can inhibit endometrial epithelial cell pyroptosis in endometritis by suppressing the IKBKE/NLRP3 signaling axis.
本文研究了黄芩炭（Cb-SR）对子宫内膜炎小鼠子宫内膜上皮细胞焦亡的抑制作用及其与IKBKE/NLRP3信号轴的相关性。通过向子宫内注射10 μL多糖（LPS，5 mg·mL⁻¹）建立子宫内膜炎小鼠模型，将小鼠随机分为模型组（LPS）、Cb-SR低剂量组（L-Cb-SR，0.55 g·kg⁻¹）、Cb-SR中剂量组（M-Cb-SR，1.10 g·kg⁻¹）、Cb-SR高剂量组（H-Cb-SR，2.20 g·kg⁻¹）、黄芩生药组（Cr-SR，1.63 g·kg⁻¹）和妇科千金胶囊组（FQC，0.30 g·kg⁻¹），每组10只小鼠。选取10只健康雌性小鼠，向双侧子宫注射等体积的PBS，设为假手术组。药物治疗组小鼠每天经口灌胃给予相应剂量的Cb-SR、Cr-SR、FQC或等体积的生理盐水，连续7天。末次给药30分钟后，颈椎脱臼处死每只小鼠。采用苏木精-伊红（HE）染色和透射电子显微镜观察子宫组织的组织病理学形态。免疫组织化学法检测CD38和CD138的表达。用试剂盒检测中性粒细胞中的髓过氧化物酶（MPO）值；采用酶联免疫吸附测定（ELISA）法检测白细胞介素-18（IL-18）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的分泌。免疫荧光和蛋白质印迹法分析IKBKE/NLRP3信号轴相关蛋白的表达。通过体外实验从妊娠雌性小鼠子宫组织中分离纯化小鼠子宫内膜上皮细胞（MEECs），用LPS损伤24小时，然后用含Cb-SR的血清培养。通过CCK-8法、扫描电子显微镜和蛋白质印迹法研究Cb-SR的抗子宫内膜炎作用。结果表明，Cb-SR可显著降低MPO值，减轻子宫组织损伤，抑制CD38和CD138的表达，降低IL-1β、IL-18和TNF-α水平，并抑制子宫内膜炎小鼠中与IKBKE/NLRP3信号轴相关蛋白的表达。此外，含Cb-SR的血清可减轻LPS诱导的MEECs细胞器肿胀，降低炎症因子表达，并抑制IKBKE/NLRP3信号轴相关蛋白的表达。这些结果表明，Cb-SR可通过抑制IKBKE/NLRP3信号轴来抑制子宫内膜炎中子宫内膜上皮细胞的焦亡。
