基于TAS2R38探究黄连“苦寒败胃”的生物基础

目的基于苦味受体38(bitter taste receptor,TAS2R38)探讨久服、误服黄连导致“苦寒败胃”的生物基础。方法ICR小鼠随机分为5组,分别为空白对照组、黄连低(0.5 g/kg)/中(1.5 g/kg)/高(4.5 g/kg)剂量组和苯甲地那铵组(0.225 g/kg),每组12只。各组小鼠分别灌胃给予相应的药物,持续8周。于第2、4、8周处理动物,RT-qPCR检测小鼠胃肠TAS2R38 mRNA表达水平;观察小鼠胃排空、肠推进;ELISA测定小鼠胃泌素和胆囊收缩素;HE染色评估胃肠黏膜损伤程度。结果与空白对照组相比:苯甲地那铵和黄连均显著增加TAS2R38 mRNA表达水平,其中黄连组呈剂量依赖性(P<0.01);苯甲地那铵延缓胃排空(P<0.05),黄连在第2周时促进胃排空(P<0.01),但在第4、8周则延缓胃排空,特别是中、高剂量组(P<0.05);苯甲地那铵与黄连均呈时间依赖性地抑制肠蠕动,尤以黄连高剂量组表现明显(P<0.01);苯甲地那铵与黄连均抑制胃泌素的分泌、促进胆囊收缩素的分泌(P<0.05,P<0.01);HE染色显示苯甲地那铵和黄连均可损伤胃肠组织。结论长期使用黄连可通过激动TAS2R38,影响小鼠胃肠动力,调节胃肠激素分泌,加重胃肠组织损伤,可能是其“苦寒败胃”的生物基础之一。

Exploring the Biological Basis of Huanglian(Coptidis Rhizoma)of"KuHanBaiWei"Based on TAS2R38

Objective Based on the bitter taste receptor 38(TAS2R38)to explore the biological basis of"KuHanBaiWei"caused by long-term or mistaken intake of Huanglian(Coptidis Rhizoma).Methods ICR mice were randomly divided into five groups,including the control group,the Huanglian(Coptidis Rhizoma)in low-dose group(0.5 g/kg),middle-dose group(1.5 g/kg),high-dose group(4.5 g/kg)and denatonium benzoate group(0.225 g/kg),with 12 mice in each group.All mice were treated with the corresponding drugs by gavage for 8 weeks.The mice were killed at 2nd,4th,and 8th weeks,RT-qPCR was performed to examine the mRNA expression of TAS2R38 in gastrointestinal tract of mice;the gastric emptying and small intestinal propulsion of mice were observed;ELISA was used to detect gastrin and cholecystokinin in mice;HE staining was used to assess the gastrointestinal mucosal lesions.Results Denatonium benzoate group and Huanglian(Coptidis Rhizoma)group could significantly increase the mRNA expression of TAS2R38 in a dose-dependent manner compared with the control group(P<0.01);denatonium benzoate delayed gastric emptying(P<0.05);Huanglian(Coptidis Rhizoma)could promote gastric emptying at the 2nd week(P<0.01),and delay gastric emptying at the 4th and 8th week,especially in the middle-dose group and the high-dose group(P<0.05);both denatonium benzoate and Huanglian(Coptidis Rhizoma)inhibited intestinal peristalsis in a time-dependent manner,especially in the high-dose group(P<0.01);both denatonium benzoate and Huanglian(Coptidis Rhizoma)inhibited the secretion of gastrin and promoted the secretion of cholecystokinin(P<0.05,P<0.01);HE staining showed that both denatonium benzoate and Huanglian(Coptidis Rhizoma)could damage gastrointestinal tissue.Conclusion Long-term use of Huanglian(Coptidis Rhizoma)can affect gastrointestinal motility,regulate gastrointestinal hormone secretion and aggravate gastrointestinal injury in mice by activating TAS2R38,which may be one of the biological basis of"KuHanBaiWei".