| 韩世诚,王婷婷,张一昕,王茜,柴天.楮实子预处理对对乙酰氨基酚致L02细胞损伤的保护作用[J].中国医药导报,2024,21(1):34-38 |
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| 楮实子预处理对对乙酰氨基酚致L02细胞损伤的保护作用 |
| Protective effect of fructus broussonetiae on APAP-induced injury of L02 cells |
| 收稿日期：2023-05-08 |
| DOI：10.20047/j.issn1673-7210.2024.01.06 |
| 关键词: 对乙酰氨基酚 楮实子 药物性肝损伤 线粒体功能 内质网应激 |
| Key Words: |
| 基金项目:河北省中医药管理局科研计划项目（2022362） |
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| 摘要:目的 探讨楮实子对对乙酰氨基酚（APAP）引起的L02细胞损伤的保护作用。 方法 CCK-8法测定不同浓度（0.1、0.3、1.0、3.0、10.0 mg/ml）楮实子水提物溶液对正常L02细胞的存活率。L02细胞APAP染毒后，取对数生长期的L02细胞，分为正常组、APAP组（40 mmol/L）、楮实子组（1 mg/ml），采用CCK-8法测定细胞存活率，采用DCFA-DH法测定细胞内活性氧（ROS）水平。Hoechst 33258染色观察细胞凋亡；罗丹明123染色检测L02细胞内线粒体膜电位表达情况。Western blot法检测细胞JNK、p-JNK、78GRP78、CHOP的表达水平。 结果 与0.1 mg/ml楮实子水提物比较，0.3、1.0、3.0、10.0 mg/ml楮实子水提物细胞活性降低，差异有统计学意义（P＜0.01）。与正常组比较，APAP组细胞存活率下降，细胞凋亡平均荧光强度值升高（P＜0.05）；与APAP组比较，1 mg/ml楮实子水提物细胞存活率升高，细胞凋亡平均荧光强度值降低（P＜0.05）。APAP组ROS荧光强度值高于正常组，差异有高度统计学意义（P＜0.01）；楮实子组ROS荧光强度值低于APAP组，差异有高度统计学意义（P＜0.01）。APAP组线粒体膜电位平均荧光强度值低于正常组，差异有高度统计学意义（P＜0.01）；楮实子组线粒体膜电位平均荧光强度值高于APAP组，差异有高度统计学意义（P＜0.01）。各组JNK蛋白表达水平比较，差异无统计学意义（P＞0.05）。与正常组比较，APAP组p-JNK、GRP78、CHOP蛋白表达水平升高（P＜0.01）；与APAP组比较，楮实子组p-JNK、GRP78、CHOP蛋白表达水平降低（P＜0.01）。 结论 楮实子可能通过抑制JNK活化，修复线粒体膜通透性，抑制内质网应激，减少ROS含量，抑制细胞凋亡，发挥保护APAP致L02细胞损伤的作用。 |
| Abstract:Objective To investigate the protective effect of fructus broussonetiae on L02 cell damage induced by acetaminophen (APAP). Methods The survival rate of normal L02 cells was determined by CCK-8 method at different concentrations (0.1, 0.3, 1.0, 3.0, 10.0 mg/ml). After the L02 cells were infected with APAP, the L02 cells with logarithmic growth stage were selected and divided into normal group, APAP group (40 mmol/L) and fructus broussonetiae group (1 mg/ml). The cell survival rate was determined by CCK-8 method, and the intracellular ROS levels were determined by DCFA-DH method. Apoptosis was observed by Hoechst 33258 staining. The expression of mitochondrial membrane potential in L02 cells was detected by Rhodamine 123 staining. The expression levels of JNK, p-JNK, 78GRP78, and CHOP were detected by Western blot. Results Compared with 0.1 mg/ml fructus broussonetia water extract, the cell activity of 0.3, 1.0, 3.0, and 10.0 mg/ml fructus broussonetia water extract decreased, and the difference was highly statistically significant (P＜0.01). Compared with normal group, the cell survival rate and the average fluorescence intensity of apoptosis of APAP group were decreased (P＜0.05). Compared with APAP group, the survival rate of 1 mg/ml fructus broussonetia water extract increased, and the average fluorescence intensity of apoptosis decreased (P＜0.05). The ROS fluorescence inten- sity of APAP group was higher than that of normal group, and the differences were highly statistically significant (P＜0.01). The ROS fluorescence intensity of fructus broussonetiae seed group was lower than that of APAP group, and the differences were highly statistically significant (P＜0.01). The average fluorescence intensity of mitochondrial membrane potential in APAP group was lower than that in normal group, and the difference was highly statistically significant (P＜0.01). The average fluorescence intensity of mitochondrial membrane potential in fructus broussonetia group was higher than that in APAP group, and the difference was highly statistically significant (P＜ 0.01). There was no significant difference in JNK protein expression among all groups (P＞0.05). Compared with normal group, the expression levels of P-JNK, GRP78, and CHOP protein in APAP group were increased (P＜0.01). Compared with APAP group, the protein expression levels of P-JNK, GRP78 and CHOP in fructus broussonetiae group were decreased (P＜0.01). Conclusion Fructus broussonetiae mitigates liver injury by inhibiting JNK，restoring mitochondrial membrane permeability，suppressing endoplasmic reticulum stress，lowering ROS levels，and inhibiting apoptosis. |
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