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徐大志,温如燕,李相玲,余浚龙.双黄连冻干粉成分分析及其与头孢拉定合用对人肝细胞功能的影响[J].湖南中医药大学学报,2022,42(9):1451-1456[点击复制] |
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| 双黄连冻干粉成分分析及其与头孢拉定合用对人肝细胞功能的影响 |
| 徐大志,温如燕,李相玲,余浚龙 |
| (大庆市人民医院, 黑龙江 大庆 163316;广西中医药大学, 广西 南宁 530200) |
| 摘要: |
| 目的 检测双黄连冻干粉中的成分含量并探讨双黄连冻干粉及其成分与头孢拉定合用对人肝细胞（HL-7702）功能的影响。方法 应用高效液相色谱（high performance liquid chromatography,HPLC）检测双黄连冻干粉主要成分黄芩苷、汉黄芩素、绿原酸、连翘苷的含量。取双黄连冻干粉以及双黄连冻干粉中各主要成分单独或与头孢拉定合用对HL-7702分别分组进行培养。酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测培养后细胞上清液中谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)的含量；HPLC检测细胞中二磷酸腺苷（adenosine diphosphate,ADP）、三磷酸腺苷（adenosine triphosphate,ATP）的含量；琼脂糖凝胶电泳检测HL-7702细胞环氧化酶2（cyclooxygenase 2,COX-2）、血红素加氧酶-1(heme oxygenase-1,HO-1）的表达水平。结果 注射用双黄连冻干粉中，黄芩苷含量最高，汉黄芩素含量最低。与对照组相比，高剂量黄芩苷组、连翘苷组、汉黄芩素组人肝细胞（HL-7702）中AST、ALT表达下降（P＜0.05，P＜0.01），绿原酸+头孢拉定组（0.046 mg/mL）与连翘苷+头孢拉定组（0.046 mg/mL）中ALT表达增加（P＜0.05，P＜0.01），AST表达差异无统计学意义（P>0.05）；低剂量组与高剂量组中各组实验结果相似。与对照组相比，高剂量组中绿原酸组、绿原酸+头孢拉定组（0.046 mg/mL）与连翘苷+头孢拉定组（0.046 mg/mL）中ATP表达下降（P＜0.05，P＜0.01），ADP表达差异无统计学意义（P>0.05）；低剂量组中黄芩苷组ATP、ADP表达增加（P＜0.05），汉黄芩素组、黄芩苷+头孢拉定组（0.046 mg/mL）与汉黄芩素+头孢拉定组（0.046 mg/mL）中ATP、ADP表达下降（P＜0.05，P＜0.01）。与对照组相比，头孢拉定组HL-7702细胞COX-2、HO-1的表达差异无统计学意义（P>0.05），双黄连不同剂量组以及与头孢拉定合用组HO-1、COX-2表达均增加且差异有统计学意义（P＜0.05，P＜0.01）。结论 注射用双黄连冻干粉中各成分对肝细胞存在影响，其中黄芩苷影响显著，与头孢拉丁合用时增加头孢拉定对肝细胞的影响。 |
| 关键词: 双黄连冻干粉 头孢拉定 HL-7702 谷丙转氨酶 谷草转氨酶 二磷酸腺苷 三磷酸腺苷 环氧化酶2 血红素加氧酶-1 |
| DOI：10.3969/j.issn.1674-070X.2022.09.006 |
| 投稿时间：2021-12-08 |
| 基金项目:广西自然科学基金项目(2018GXNSFBA050051);广西中医药大学博士启动基金项目(2020BS011)。 |
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| Composition analysis of Shuanghuanglian freeze-dried powder and its effects on human hepatocyte function when combined with cefradine |
| XU Dazhi,WEN Ruyan,LI Xiangling,YU Junlong |
| (Daqing People's Hospital, Daqing, Heilongjiang 163316, China;Guangxi University of Chinese Medicine, Nanning, Guangxi 530200, China) |
| Abstract: |
| Objective To detect the content of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes (HL-7702) alone or in combination with cefradine. Methods High performance liquid chromatography (HPLC) was used to detect the content of baicalin, wogonin, chlorogenic acid and forsythin, the main components of Shuanghuanglian freeze-dried powder. HL-7702 cells were cultured with Shuanghuanglian freeze-dried powder and the main components of Shuanghuanglian freeze-dried powder alone or in combination with cefradine. Enzyme linked immunosorbent assay (ELISA) was used to detect the content of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the cell supernatant after culture, and HPLC was used to detect the expression level of adenosine diphosphate (ADP) and adenosine triphosphate (ATP); agarose gel electrophoresis was used to detect the expression of cyclooxygenase 2 (COX-2) and heme oxygenase-1 (HO-1) in HL-7702 cells. Results In Shuanghuanglian freeze-dried powder for injection, the content of baicalin was the highest, and the content of wogonin was the lowest. Compared with the control group, the expression of AST and ALT in human hepatocytes (HL-7702) in high-dose baicalin group, forsythin group and wogonin group decreased (P<0.05, P<0.01), while the expression of ALT in chlorogenic acid+cephradine group (0.046 mg/mL) and forsythin+cephradine group (0.046 mg/mL) increased (P<0.05, P<0.01), and the expression of AST had no statistical significance (P>0.05); the results in the low-dose group were similar to those in the high-dose group. Compared with the control group, ATP expression in chlorogenic acid group, chlorogenic acid+cephradine group (0.046 mg/mL) and forsythin+cephradine group (0.046 mg/mL) in the high-dose group decreased (P<0.05, P<0.01), and ADP expression was not significantly different (P>0.05); in the low-dose group, the expression of ATP and ADP increased in baicalin group (P<0.05), but decreased in wogonin group, baicalin+cephradine group (0.046 mg/mL) and wogonin+cephradine group (0.046 mg/mL) (P<0.05, P<0.01). Compared with the control group, the expression of COX-2 and HO-1 in HL-7702 cells in the cephradine group showed no statistical significance (P>0.05). The expression of HO-1 and COX-2 in different dosage groups of Shuanghuanglian and the group combined with cephradine increased and the difference was statistically significant (P<0.05, P<0.01). Conclusion The components of Shuanghuanglian freeze-dried powder for injection have effects on hepatocytes, of which baicalin has a significant effect, and the effects of cefradine on hepatocytes are increased when used in combination with cefradine. |
| Key words: Shuanghuanglian freeze-dried powder cefradine HL-7702 alanine aminotransferase aspartate aminotransferase adenosine diphosphate adenosine triphosphate cyclooxygenase 2 heme oxygenase-1 |
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