3-甲基腺嘌呤腹腔注射干预大鼠胃窦组织自噬最佳药物剂量与用药时间探讨

Optimal dose and time of intraperitoneal injection of 3-MA for intervention of gastric antrum autophagy in rats

目的:探讨3-甲基腺嘌呤(3-MA)腹腔注射干预大鼠胃窦组织自噬的最佳药物剂量及用药时间。方法:将21只健康雄性SD大鼠随机分为七组:A组3-MA 10 mg/kg组、B组3-MA 15 mg/kg组、C组3-MA 30 mg/kg组,D组PBS组,E组空白对照组,F组3-MA 30 mg/kg 3 d组、G组3-MA 30 mg/kg 1 d组,每组3只。3-MA分别按照大鼠体重以剂量10、15、30 mg/kg进行腹腔注射,D组大鼠给予等量PBS溶液腹腔注射,空白对照组不予处理,A、B、C、D、E组干预15 d,F组干预3 d,G组干预1 d。干预结束后取胃窦组织,用Western blot法检测其LC3、P62的表达。结果:LC3Ⅱ/LC3Ⅰ比值大小排序为:D、E组>A组>B组>C组(均P<0.05),G组>C组>F组(均P<0.05);P62相对表达量排序为:C组>B组>A组>D、E组(均P<0.05),C组>G组>F组(均P<0.05)。D、E组LC3Ⅱ/LC3Ⅰ比值和P62相对表达量比较均无统计学差异(均P>0.05)。结论:3-MA腹腔注射可以抑制胃窦组织自噬,其中30 mg/kg为3-MA腹腔注射抑制大鼠胃窦组织自噬的最佳药物剂量,腹腔注射干预15 d自噬抑制效果最明显。

Objective:To explore the optimal dose and time of intraperitoneal injection of 3-MA for intervention of gastric antrum autophagy in rats.Methods:21 healthy male SD rats were randomly divided into 7 groups:A group(3-MA 10 mg/kg),B group(3-MA 15 mg/kg),C group(3-MA 30 mg/kg),D group(PBS),E group(blank control),F group(3-MA 30 mg/kg),G group(3-MA 30 mg/kg),3 rats in each group.3-MA was injected intraperitoneally with doses of 10,15,and 30 mg/kg according to the body weight of rats.The rats in group D were injected intraperitoneally with the same amount of PBS.The blank control group was not treated.A,B,C,D,E groups were intervened for 15 days,F group was intervened for 3 days,and G group was intervened for 1 day.After the intervention,the gastric antrum tissue was taken and the expression of LC3 and p62 was detected by Western blot.Results:The order of LC3Ⅱ/LC3Ⅰratio was group D,E>group A>group B>group C(all P<0.05),group G>group C>group F(all P<0.05).The relative expression of P62 was sorted as follows:group C>group B>group A>group D,E(all P<0.05),group C>group G>group F(all P<0.05).There was no significant difference in the ratio of LC3Ⅱ/LC3Ⅰand the relative expression of P62 between group D and group E(all P>0.05).Conclusion:Intraperitoneal injection of 3-MA can inhibit autophagy of gastric antrum tissue,and 30 mg/kg is the optimal dose in rats,and the optimal intervention time is 15 days.