柚皮素对溃疡性结肠炎小鼠肠组织中miR-30a-3pPTEN信号轴表达的影响

The effect of naringenin on the expression of miR-30a-3p/PTEN signal axis in the intestinal tissue of mice with ulcerative colitis

目的 探讨柚皮素对溃疡性结肠炎小鼠肠组织中miR-30a-3p/PTEN信号轴表达的影响。方法 选取60只健康雄性BALB/c小鼠,按随机数字表法均分为阴性对照组、模型组、阳性对照组、柚皮素低剂量组、柚皮素中剂量组和柚皮素高剂量组,每组各10只。用DSS灌胃法制备BALB/c小鼠溃疡性结肠炎模型,阴性对照组和模型组灌胃给予生理盐水(0.2 mL/10g),阳性对照组灌胃给予泼尼松龙(2 mg/kg),柚皮素低、中、高剂量组分别灌胃给予柚皮素(剂量依次为50、100、200 mg/kg),持续给予7 d。计算疾病活动度评分(DAI),对BALB/c小鼠肠组织进行形态学检查,同时采用酶联免疫吸附(ELISA)法检测肠组织中白细胞介素-4(IL-4)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)水平,实时荧光定量PCR(RT-PCR)法及Elisa法分别检测肠组织miR-30a-3p、PTEN、PI3K、Akt和Caspase-3 mRNA及蛋白水平。结果 模型组小鼠结肠有部分肠壁凹陷,形成溃疡,周围组织水肿明显,部分细胞坏死、脱落,炎性细胞浸润;与模型组比较,阳性对照组和各柚皮素剂量组小鼠结肠组织病理损伤明显改善。与阴性对照组比较,其余各组小鼠肠组织DAI、IL-4、TNF-α、PTEN和Caspase-3 mRNA及蛋白升高,IL-10、miR-30a-3p、PI3K和Akt mRNA及蛋白降低(均P<0.05);与模型组比较,阳性对照组和各柚皮素剂量组小鼠肠组织DAI、IL-4、TNF-α、PTEN和Caspase-3 mRNA及蛋白降低,IL-10、miR-30a-3p、PI3K和Akt mRNA及蛋白升高,各柚皮素剂量组效应呈剂量反应关系(均P<0.05);柚皮素高剂量组与阳性对照组各指标比较差异无统计学意义(P>0.05)。结论 柚皮素可以降低溃疡性结肠炎小鼠的结肠炎性,减轻病理损伤,其机制可能与调控miR-30a-3p/PTEN信号轴有关。

Objective To investigate the effect of naringenin on the expression of miR-30a-3p/PTEN signal axis in the intestinal tissue of mice with ulcerative colitis.Methods Ulcerative colitis model of BALB/c mice was prepared by DSS gavage method.The negative control group and model group were gavaged with normal saline(0.2 mL/10g),and the positive control group was gavaged with prednisolone(2 mg/kg).The low,middle and high-dose naringenin groups were given naringenin by intragastric administration(the doses were 50,100,and 200mg/kg)for 7 days.The disease activity score(DAI)was calculated,morphological examination of intestinal tissue in BALB/c mice performed evaluated,and Elisa was used to detect the levels of IL-4,IL-10,and TNF-αin intestinal tissue,RT-PCR and Elisa method measure the levels of miR-30a-3p,PTEN,PI3K,Akt and Caspase-3 mRNA and protein in intestinal tissue respectively.Results The mouse colon in the model group had partial intestinal wall depression,formation of ulcers,obvious edema in the surrounding tissues,necrosis and shedding of some cells,and inflammatory cell infiltration.The damage of colon tissue pathology of the mice in the positive control group and each naringenin dosage group was significantly improved compared with the model group.Compared with the negative control group,DAI,IL-4,TNF-α,PTEN and Caspase-3 mRNA and protein in the intestinal tissues of the other groups of mice increased,and IL-10,miR-30a-3p,PI3K and Akt mRNA and protein decreased(P<0.05).Compared with the model group,the DAI,IL-4,TNF-α,PTEN and Caspase-3 mRNA and protein of mice in the positive control group and each naringenin dose group decreased,IL-10,miR-30a-3p,PI3K and Akt mRNA and protein increased,and the effects of each naringenin dose group showed a dose-response relationship(P<0.05).There was no significant difference in indicators between the high-dose naringenin group and the positive control group(P>0.05).Conclusion Naringenin can reduce colitis and pathological damage in mice with ulcerative colitis.The mechanism may be related to the regulation of miR-30a-3p/PTEN signal axis.