桂枝茯苓胶囊及其主要成分对原发性痛经模型大鼠肠道菌群的影响

Effects of Guizhi Fuling Capsules and Its Main Components on Intestinal Flora of Primary Dysmenorrhea Model Rats

目的:探讨桂枝茯苓胶囊及其主要成分(芍药苷、丹皮酚、苦杏仁苷)对原发性痛经模型大鼠肠道菌群的影响。方法:将雌性SD大鼠随机分为正常组、模型组、胶囊组(桂枝茯苓胶囊内容物,1000 mg/kg)、芍药苷组(15.0 mg/kg)、丹皮酚组(10.3 mg/kg)和苦杏仁苷组(12.1 mg/kg),每组6只。除正常组外,其余各组大鼠均于背部皮下注射苯甲酸雌二醇和腹腔注射缩宫素建立原发性痛经模型。于皮下注射苯甲酸雌二醇的第4天起,正常组大鼠灌胃等体积生理盐水,模型组大鼠灌胃等体积0.5%羧甲基纤维素钠溶液,各给药组大鼠灌胃相应药物,每天1次,连续7 d。在检测各组大鼠扭体次数和子宫组织中一氧化氮(NO)、丙二醛(MDA)含量的基础上,采用气相色谱法检测其结肠内容物中乙酸、丙酸、丁酸等短链脂肪酸(SCFAs)的含量;以多样性指数为指标,采用细菌基因组重复序列-聚合酶链式反应(Rep-PCR)和肠杆菌基因组间重复序列-聚合酶链式反应(Eric-PCR)技术评价大鼠粪便中肠道菌群的多样性。结果:与正常组比较,模型组大鼠扭体次数显著增多,子宫组织中NO、MDA含量均显著升高,结肠内容物中乙酸、丙酸、丁酸及总SCFAs含量均显著降低(P<0.05或P<0.01);肠道菌群DNA电泳条带数量明显减少,大部分条带亮度明显减弱,且给药1 h时其多样性指数(Rep-PCR和Eric-PCR法,下同)均显著降低(P<0.05或P<0.01)。与模型组比较,胶囊组、芍药苷组、丹皮酚组大鼠的扭体次数均显著减少,胶囊组、丹皮酚组大鼠子宫组织中NO含量以及胶囊组、芍药苷组、丹皮酚组MDA含量均显著降低(P<0.05或P<0.01);胶囊组大鼠结肠内容物中丙酸及总SCFAs含量,芍药苷组乙酸、丙酸、丁酸及总SCFAs含量以及丹皮酚组丙酸、丁酸及总SCFAs含量均显著升高,而芍药苷组异戊酸含量显著降低(P<0.05或P<0.01);各给药组大鼠肠道菌群DNA电泳条带及亮度均有不同程度的改变,且给药1 h时胶囊组和芍药苷组大鼠肠道菌群的多样性指数均显著升高,而丹皮酚组和苦杏仁苷组大鼠肠道菌群的上述多样性指数均显著降低(P<0.05或P<0.01)。结论:桂枝茯苓胶囊可显著减少原发性痛经模型大鼠的扭体次数,降低其子宫组织中NO、MDA的含量;同时,该药还可调节大鼠结肠内容物中SCFAs含量和肠道菌群多样性。上述作用可能与该药中的芍药苷和丹皮酚等成分有关。

OBJECTIVE:To investigate the effects of Guizhi fuling capsules and its principal components(paeoniflorin,paeonol and amygdalin)on the intestinal flora of primary dysmenorrhea model rats.METHODS:Female SD rats were randomly divided into normal group,model group,capsule group(Guizhi fuling capsule,1000 mg/kg),paeoniflorin group(15.0 mg/kg),paeonol group(10.3 mg/kg)and amygdalin group(12.1 mg/kg),with 6 rats in each group.Except for normal group,other groups were given estradiol benzoate subcutaneously on the back of rats and oxytocin intraperitoneally to induce primary dysmenorrhea model.From the 4th day after subcutaneous injection of estradiol benzoate,normal group was given constant volume of normal saline intragastrically;model group was given constant volume of 0.5%CMC-Na solution intragastrically;administration groups were given relevant medicine intragastrically,once a day,for consecutive 7 days.The writhing times and the contents of NO and MDA in uterus tissue of rats were determined,and then the contents of short-chain fatty acids(SCFAs)such as acetic acid,propionic acid,butyric acid in colonic contents were detected by GC method.Using diversity index as index,Rep-PCR and Eric-PCR were used to evaluate the diversity of intestinal flora in feces of rats.RESULTS:Compared with normal group,the writhing times of rats were increased significantly in model group;the contents of NO and MDA in uterus were increased significantly,while the contents of acetic acid,propionic acid and butyric acid in colonic contents and total content of SCFAs were decreased significantly(P<0.05 or P<0.01);the number of DNA electrophoresis bands of intestinal flora was significantly reduced,the brightness of most bands was significantly reduced,and the diversity indexes(by Rep-PCR and Eric-PCR method,hereinafter)1 h after administration were significantly reduced(P<0.05 or P<0.01).Compared with model group,writhing times of rats were decreased significantly in capsule group,paeoniflorin group and paeonol group;the contents of NO in uterus of rats in capsule group and paeoniflorin group as well as the contents of MDA in capsule group,paeoniflorin group and paeonol group were decreased significantly(P<0.05 or P<0.01);the propionic acid content and total content of SCFAs in colon of rats in capsule group,the contents of acetic acid,propionic acid and butyric acid,total content of SCFAs in paeoniflorin group as well as the contents of propionic acid and butyric acid,total content of SCFAs in paeonol group were increased significantly;the content of isovaleric acid was decreased significantly in paeoniflorin group(P<0.05 or P<0.01);DNA electrophoresis bands and its brightness of intestinal flora changed to different extents in administration groups,and the diversity indexes of intestinal flora 1 h after administration were increased significantly in capsule group and paeoniflorin group,while those indexes were decreased significantly in paeonol group and amygdalin group(P<0.05 or P<0.01).CONCLUSIONS:Guizhi fuling capsules can significantly reduce writhing times and the contents of NO and MDA in uterus of primary dysmenorrhea model rats.At the same time,the capsules also can regulate SCFAs content in colonic contents and intestinal flora diversity of rats.The above effects may be related to paeoniflorin and paeonol in the capsules.