葛根素对慢性酒精中毒小鼠结肠组织Caj al间质细胞和C-kit蛋白表达的影响

摘要

Objective To explore the protective effects of puerarin on the colon tissue of the mice with chronic alcoholism through observing the changes of quantity and morphology of interstitial cells of Cajal (ICC)and the expression of c-kit protein in colon tissue of chronic alcoholism mice.Methods 24 healthy BALB/C mice were randomly divided into saline control group (SC group ), chronic alcoholic intoxication group (CAI group ) and puerarin pretreatment group(PUE group).The models of chronic alcoholism mice were established in CAI group and PUE group. The intestinal transmission rate was tested by measuring the Indian ink advancing length;immunofluorescence and transmission electron microscope (TEM)were applied to detect the distribution, quantity and ultrastructure of ICC;Western blotting method was used to analyze the C-kit protein expression.Results The intestinal transmission rate in PUE group was significant higher than that in CAI group(P<0.05),but was still lower than that in SC group (P<0.05);compared with CAI group,the number of ICC in PUE group was increased (P<0.05),but there was no significant difference between SC group and PUE group(P>0.05).Compared with SC group,the ultrastructure of ICC in CAI group had obvious changes,the organelles were decreased,and the mitochondrion was swelling.But those changes were reversed in PUE group and the number of mitochondria was increased.The expression of C-kit protein in PUE tissue was higher than that in CAI group (P<0.05),but there was no significant difference compared with SC group.Conclusion Puerarin has a repair or reverse effect on the changes of the number,ultrastructure of ICC and C-kit protein expression caused by alcohol.%目的：通过检测慢性酒精中毒及葛根素预处理后的小鼠结肠 Caj al 间质细胞（ICC）形态、数量以及C-kit蛋白表达的变化，探讨葛根素对酒精中毒小鼠结肠组织的保护作用。方法：选取健康雄性 BALB／C 小鼠24只，随机分为盐水对照组、慢性酒精中毒组和葛根素预处理组，每组8只。慢性酒精中毒组和葛根素预处理组小鼠建立慢性酒精中毒动物模型。通过测量印度墨汁推进长度测定肠道传输速率，应用免疫荧光和透射电镜技术检测 ICC分布、数量以及超微结构的改变，采用 Western blotting方法分析 C-kit蛋白的表达。结果：葛根素预处理组肠道传输速率较慢性酒精中毒组明显增加（P＜0.05），但仍低于盐水对照组（P＜0.05）；葛根素预处理组结肠内 ICC的数量较慢性酒精中毒组增多（P＜0.05），与盐水对照组比较差异无统计学意义（P＞0.05）；与盐水对照组比较，慢性酒精中毒组小鼠结肠 ICC的超微结构发生明显改变，线粒体等细胞器明显减少，葛根素预处理组小鼠结肠 ICC的超微结构基本恢复正常，线粒体等细胞器明显增加；葛根素预处理组小鼠结肠组织中C-kit蛋白表达水平明显高于慢性酒精中毒组（P＜0.05），与盐水对照组比较差异无统计学意义（P＞0.05）。结论：葛根素干预对酒精造成的Caj al间质细胞数量、超微结构及C-kit表达的改变有一定的修复或逆转作用。