植物乳杆菌联合魔芋葡甘聚糖改善小鼠便秘的作用和机制研究

Study on the effect and mechanism of Lactobacillus plantarum combined with konjac glucomannan in improving constipation in mice.

目的探讨植物乳杆菌联合魔芋葡甘聚糖改善小鼠便秘的作用和机制研究。方法根据实验目的将60只雄性昆明小鼠分为对照组、便秘组、联合治疗组,每组各20只。对照组给予皮下注射生理盐水,便秘组给予皮下注射盐酸洛哌丁胺诱导便秘模型,持续6 d,联合治疗组在小鼠诱导便秘模型的6 d,饮食中添加LP-KSFY06,0.5×107CFU/kg,魔芋葡甘聚糖500 mg/kg。比较实验期间第6、8和10天的小鼠体重。通过称重分析每组小鼠的粪便重量和粪便含水量。收集小肠进行活性炭通过率分析,并比较排黑便时间。通过力传感器检测小鼠离体结肠平滑肌的收缩频率。通过蛋白质印迹分析小鼠结肠组织中c-Kit、TRPV1、GDNF、5-HT4R和SERT的蛋白表达。通过试剂盒检测小肠组织中髓过氧化物酶(MPO)、一氧化氮(NO)、谷胱甘肽(GSH)和丙二醛(MDA)的水平。结果第6天,3组小鼠体重比较,差异无统计学意义(P>0.05),第8、10天,便秘组较对照组小鼠体重降低(P<0.05),联合治疗组较便秘组体重升高(P<0.05)。便秘组较对照组粪便重量和粪便含水量降低(P<0.05),联合治疗组较便秘组粪便重量和粪便含水量升高(P<0.05)。便秘组较对照组活性炭推进率降低,第一次排出黑便时间延长(P<0.05),联合治疗组较便秘组活性炭推进率升高,第一次排黑便时间缩短(P<0.05),3组小肠长度比较差异无统计学意义(P>0.05)。便秘组较对照组近端结肠纵向SM和远端结肠环形SM的收缩频率降低(P<0.05),联合治疗组较便秘组收缩频率升高(P<0.05)。便秘组较对照组c-Kit、TRPV1的蛋白表达升高(P<0.05),GDNF、5-HT4R和SERT蛋白表达降低(P<0.05),联合治疗组较便秘组c-Kit、TRPV1的蛋白表达降低(P<0.05),GDNF、5-HT4R和SERT蛋白表达升高(P<0.05)。便秘组较对照组MPO、NO、MDA含量升高,GSH含量降低(P<0.05),联合治疗组较便秘组MPO、NO、MDA含量降低,GSH含量升高(P<0.05)。结论植物乳杆菌联合魔芋葡甘聚糖促进便秘小鼠的肠道蠕动,改善便秘症状,是可用于预防或治愈便秘的潜在药物。

Objective To investigate the effect and mechanism of Lactobacillus plantarum combined with konjac glucomannan to improve constipation in mice.Methods According to the experimental objectives,sixty male Kunming(KM)mice were divided into control group,constipation group,and combined treatment group,each group 20 mice.The control group was given a subcutaneous injection of normal saline,and the constipation group was given subcutaneous injection of loperamide hydrochloride to induce a constipation model for 6 days.The combined treatment group was given LP-KSFY06,0.5×107 CFU/kg,konjac Glucomannan 500 mg/kg in the diet when the constipation model was induced.The body weight of mice on the 6th,8th and 10th day during the experiment was compared.The fecal weight and fecal water content of each group of mice were analyzed by weighing.The small intestines were collected for analysis of the passing rate of activated carbon,and the time to defecation was compared.The protein expression of c-Kit,TRPV1,GDNF,5-HT4R and SERT in mice colon tissue were analyzed by Western blotting.The levels of myeloperoxidase(MPO),nitric oxide(NO),glutathione(GSH),malondialdehyde(MDA)in the small intestine tissue through the kit were detected.Results There was no difference in the weight of mice in each experimental group on the 6th day(P>0.05).The weight of mice in the constipation group on the 8th and 10th day were lower than those of the control group(P<0.05),and the weight of the combined treatment group were higher than those of the constipation group(P<0.05).The stool weight and fecal water content of the constipation group were lower than those of the control group(P<0.05),and the fecal weight and fecal water content of the combination treatment group were higher than those of the constipation group(P<0.05).Compared with the control group,the activated carbon propulsion rate of the constipation group was lower than that of the control group,and the first black stool discharge time was prolonged(P<0.05).Compared with the constipation group,the activated carbon propulsion rate of the combined treatment group was higher than that of the constipation group,and the first black stool discharge time was shortened(P<0.05).There was no difference in length between three groups(P>0.05).The contraction frequency of the longitudinal SM of the proximal colon and the circular SM of the distal colon in the constipation group were lower than thsoe of the control group(P<0.05),and the contraction frequency of the combination treatment group were higher than those of the constipation group(P<0.05).Compared with the control group,c-Kit and TRPV1 protein expressions were increased in the constipation group(P<0.05),and the GDNF,5-HT4R and SERT protein expressions were decreased(P<0.05).Compared with the constipation group,the protein expression of c-Kit and TRPV1 were decreased in the combination treatment group(P<0.05),and the protein expression of GDNF,5-HT4R and SERT was increased(P<0.05).Compared with the control group,the MPO,NO,MDA content in the constipation group were increased,and the GSH content were decreased(P<0.05).Compared with the constipation group,the MPO,NO,MDA content in the combined treatment group were decreased,and the GSH content were increased(P<0.05).Conclusion Lactobacillus plantarum combined with konjac glucomannan can promote the intestinal motility of constipated mice and improve the symptoms of constipation.It is a potential drug that can be used to prevent or cure constipation.