小檗碱和育亨宾对LPS诱导的小鼠肠道损伤和肠上皮细胞增殖抑制的影响

Effect of berberine and yohimbine on impaired enterocyte proliferation and intestinal injury in LPS-challenged mice

目的:观察小檗碱(Ber)和育亨宾(Y)对LPS诱导的小鼠肠道损伤和肠上皮细胞增殖抑制的影响。方法:雄性BALB/c小鼠随机分为对照组(control)、脂多糖组(LPS)、小檗碱组(Ber+LPS)、小檗碱与育亨宾合剂组(Ber+Y+LPS)、育亨宾组(Y+LPS)、小檗碱组(Ber)、小檗碱和育亨宾合剂组(Ber+Y)及育亨宾组(Y)。分别予以双蒸水、Ber(50mg/kg)、Ber(50mg/kg)+Y(2mg/kg)、Y(2mg/kg)灌胃,1time/d,连续3d,于实验第3d灌胃后1h,腹腔注射生理盐水或LPS(18mg/kg,0.2mL/10g)。观察各组小鼠肠组织形态学、肠黏膜重量和绒毛高度的改变;酶联免疫吸附实验(ELISA)测定肠组织二胺氧化酶(DAO)含量;免疫组化染色方法分析各组肠组织增殖细胞核抗原(PCNA)的表达。结果:LPS组小鼠肠腔显示炎性渗出、出血;肠损伤评分明显高于对照组;LPS组肠黏膜重量、肠绒毛高度、肠组织DAO含量、肠上皮细胞PCNA表达明显低于对照组。与LPS组比较,Ber组、Ber+Y组上述改变明显减轻,但两组间未见明显差异;育亨宾对LPS引起的上述指标变化无明显抑制作用。结论:Ber通过非α2肾上腺素能受体依赖的途径减轻LPS引起的肠上皮细胞增殖抑制和肠道损伤。

AIM：To evaluate the effects of oral berberine （Ber） and yohimbine （Y） in preventing intestinal damage and impaired enterocyte proliferation caused by lipopolysaccharide （LPS） in mice. METHODS：Male BALB/c mice were randomly divided into 8 groups：control,LPS,Ber ＋ LPS,Ber ＋ Y ＋ LPS,Y ＋ LPS,Ber,Ber ＋ Y and Y. The mice were administered intragastrically with distilled water （0. 1 mL/10 g）,Ber （50 mg/kg）,Ber （50 mg/kg） in combination with Y （2 mg/kg） or Y （2 mg/kg） once a day for 3 days. One hour after intragastrical treatment on the third day,LPS （18 mg/kg） or normal saline was injected intraperitoneally. Twenty hours after LPS administration,the histological changes of the intestine were observed,and injury score was assessed. The mucosal weight,villus height and the content of diamine oxidase （DAO） in the ileum were also measured. Furthermore,the proliferation of enterocyte was identified by immunohistochemical staining for proliferating cell nuclear antigen （PCNA）. RESULTS：Compared to the control mice,the mice challenged with LPS resulted in intestinal injury,including significantly increased injury score,decreased gut mucosa weight,villus height and the DAO contents in ileum. Furthermore,enterocyte proliferation was inhibited significantly 12 h after LPS challenge. Pretreatment with Ber or Ber ＋ Y significantly inhibited the intestinal injury,and attenuated the impairment of enterocyte proliferation induced by LPS. However,no significant difference in the above parameters between Ber ＋ LPS group and Ber ＋ Y ＋ LPS group was observed. Treatment with Y only did not prevent LPS-induced intestinal injury. CONCLUSION：Pretreatment with berberine remarkably reduces LPS-induced intestinal injury and the impairment of enterocyte proliferation in an alpha 2 adrenoceptor-independent manner.