Objective: To investigate whether Ershen Pill(ESP,二神丸) could alleviate the symptom of Pi(Spleen)-Shen(Kidney) yang deficiency(PSYD)-induced diarrhea in rat model and explore its anti-diarrhea mechanism. Methods: Se...Objective: To investigate whether Ershen Pill(ESP,二神丸) could alleviate the symptom of Pi(Spleen)-Shen(Kidney) yang deficiency(PSYD)-induced diarrhea in rat model and explore its anti-diarrhea mechanism. Methods: Seventy-five Sprague-Dawley rats were divided into 5 groups by a random number table, including control, positive, model, low-dose(LD) and high-dose(HD) ESP groups, 15 rats in each group. All the rats, except those in the control group, were developed PSYD induced-diarrhea based on its pathology and etiology. The rats in positive, LD and HD ESP groups were treated with Shenling Baizhu Pill(参苓白术丸), LD(1.05 g/kg) or HD(3.50 g/kg) ESP petroleum ether extract once a day for 2 weeks, respectively. Body weight change and diarrhea index were measured. The histology scores of the kidney were evaluated via hematoxylin and eosin(HE) staining. Aquaporin-3(AQP3) expression in the colon was analyzed by immunofluorescence, quantitative reverse transcription-polymerase chain reaction(qRT-PCR) and Western blot, respectively. Results: Compared with the model group, oral administration of LD and HD ESP prevented body weight loss and inhibited diarrhea after 2-week treatment(P<0.05). Kidney deterioration was impeded, and the histology score in LD and HD ESP groups were 8.2 and 10.5, respectively, which were both higher than those in the model group(P<0.05). In addition, ESP treatment alleviated rat colitis, and HD ESP significantly improved the AQP3 positive staining intensity in the colon tissue compared with the model group. The result from Western blot revealed that AQP3 protein synthesis in colon tissue of LD and HD ESP groups increased by 2.1-and 5.9-fold compared with the model group(P<0.05). qR T-PCR result showed that AQP3 gene expression in the HD ESP group was also up-regulated by 2.5-fold normalized to the model group(P<0.05). Conclusion: ESP extract effectively alleviates the symptoms of PSYD and relieves PSYD-induced diarrhea by improving AQP3 synthesis in the colon.展开更多
目的 GC-MS分析二神丸中补骨脂和肉豆蔻的炮制对其4种组合挥发性成分的影响.方法 采用水蒸气蒸馏法提取挥发油,气相色谱条件为毛细管色谱柱（HP-5 mS 5％ Phenyl methyl siloxane,30 mm ×0.25 mm ×0.25 μm）,进样口温度为250...目的 GC-MS分析二神丸中补骨脂和肉豆蔻的炮制对其4种组合挥发性成分的影响.方法 采用水蒸气蒸馏法提取挥发油,气相色谱条件为毛细管色谱柱（HP-5 mS 5％ Phenyl methyl siloxane,30 mm ×0.25 mm ×0.25 μm）,进样口温度为250℃,程序升温,载气为氦气;质谱条件为电子轰击电离（能量为70 eV）,四级杆温度为150℃,离子源温度为230℃.结果 4种组合分别检测出77、109、89和70种化合物,鉴定出了其中78种化合物的结构,占其挥发油总量的97.20％～98.87％.结论 组合Ⅳ（盐补骨脂＋煨肉豆蔻）中补骨脂酚和黄樟醚的量最低,挥发性成分的总量减少,说明补骨脂和肉豆蔻炮制后入药可降低毒性.展开更多
目的通过高效液相-飞行时间质谱(HPLC-FOF/MS)联用技术,对中药复方二神丸中的主要化学成分进行有效的鉴定。方法色谱分离采用资生堂Capcell Pak C_(18)反相柱(3.0 mm×100 mm,3μm),流动相组成分别为0.1%甲酸水溶液和乙腈,梯度洗脱...目的通过高效液相-飞行时间质谱(HPLC-FOF/MS)联用技术,对中药复方二神丸中的主要化学成分进行有效的鉴定。方法色谱分离采用资生堂Capcell Pak C_(18)反相柱(3.0 mm×100 mm,3μm),流动相组成分别为0.1%甲酸水溶液和乙腈,梯度洗脱,流速为0.6 ml/min;质谱定性采用飞行时间质谱,正离子模式扫描。结果在优化的液质联用条件下,通过飞行时间质谱鉴定出二神丸中21个成分,其中15种成分来自补骨脂,6种成分来自肉豆蔻。结论通过RRLC-TOF/MS联用技术,为鉴定二神丸中的化学成分建立起了一种快速、高效的分析方法。展开更多
基金Supported by the National Natural Science Foundation of China(No.81274085)
文摘Objective: To investigate whether Ershen Pill(ESP,二神丸) could alleviate the symptom of Pi(Spleen)-Shen(Kidney) yang deficiency(PSYD)-induced diarrhea in rat model and explore its anti-diarrhea mechanism. Methods: Seventy-five Sprague-Dawley rats were divided into 5 groups by a random number table, including control, positive, model, low-dose(LD) and high-dose(HD) ESP groups, 15 rats in each group. All the rats, except those in the control group, were developed PSYD induced-diarrhea based on its pathology and etiology. The rats in positive, LD and HD ESP groups were treated with Shenling Baizhu Pill(参苓白术丸), LD(1.05 g/kg) or HD(3.50 g/kg) ESP petroleum ether extract once a day for 2 weeks, respectively. Body weight change and diarrhea index were measured. The histology scores of the kidney were evaluated via hematoxylin and eosin(HE) staining. Aquaporin-3(AQP3) expression in the colon was analyzed by immunofluorescence, quantitative reverse transcription-polymerase chain reaction(qRT-PCR) and Western blot, respectively. Results: Compared with the model group, oral administration of LD and HD ESP prevented body weight loss and inhibited diarrhea after 2-week treatment(P<0.05). Kidney deterioration was impeded, and the histology score in LD and HD ESP groups were 8.2 and 10.5, respectively, which were both higher than those in the model group(P<0.05). In addition, ESP treatment alleviated rat colitis, and HD ESP significantly improved the AQP3 positive staining intensity in the colon tissue compared with the model group. The result from Western blot revealed that AQP3 protein synthesis in colon tissue of LD and HD ESP groups increased by 2.1-and 5.9-fold compared with the model group(P<0.05). qR T-PCR result showed that AQP3 gene expression in the HD ESP group was also up-regulated by 2.5-fold normalized to the model group(P<0.05). Conclusion: ESP extract effectively alleviates the symptoms of PSYD and relieves PSYD-induced diarrhea by improving AQP3 synthesis in the colon.
基金supported by the Major Program of the National Natural Science Foundation of China (grant nos.82192910 and 82192911)the Innovation Team and Talents Cultivation Program of the National Administration of Traditional Chinese Medicine (grant nos.ZYYCXTD-D-202207 and ZYYCXTD-C-202009)。
