五眼果水提物激活自噬预防小鼠肾结石形成的研究

Study on the Prevention of Kidney Stone Formation in Mice by Activating Autophagy with Water Extract of Choerospondias axillaris

目的探讨五眼果水提物对肾结石模型小鼠的影响及其初步机制研究。方法雄性KM小鼠随机分为六个组:正常组、模型组、阳性组(枸橼酸钾1.75 g/kg),五眼果高、中、低剂量组(40、20、10 g生药/kg),每组10只。各给药组按20 mL/kg灌胃给予相应的药物,连续30 d,正常组和模型组灌胃给予等量生理盐水。于第21天开始,除正常组外其余各组均腹腔注射乙醛酸(10 mg/mL)诱导肾结石模型,连续10 d。于第30天收集各组小鼠24 h尿液检测Ox和Ca,次日取血检测血清肌酐、尿素氮和Ca,剖取肾脏,一侧肾脏采用Western blot检测自噬相关蛋白Beclin-1、LC3B、P62和肾损伤分子(KIM-1)的表达水平;另一侧肾脏用多聚甲醛固定,通过HE染色观察肾脏结晶沉积和肾组织病理学改变。结果与模型组相比,五眼果水提物可明显降低尿钙水平(P<0.05),促进尿草酸排泄(P<0.05),明显降低血清肌酐和尿素氮水平(P<0.05),对血清Ca水平无明显影响(P>0.05)。HE染色观察结果显示五眼果水提物可明显减轻小鼠肾小管扩张和肾小管变性坏死程度,减少纤维组织增生及结晶沉积。五眼果水提物可使LC3BII/LC3BI表达水平明显升高(P<0.05),KIM-1表达水平明显降低(P<0.05)。结论五眼果水提物可通过激活自噬,减轻肾损伤,防止肾结石的形成。

Objective To investigate the effect of water extract of Choerospondias axillaris on kidney stone model mice and its preliminary mechanism.Methods Male KM mice were randomly divided into six groups:normal group,model group,positive group(potassium citrate 1.75 g/kg),high,medium and low dose groups(40,20 and 10 g crude drug/kg)of Choerospondias axillaris with 10 mice in each group.Each administration group was given corresponding drugs by gavage at 20 mL/kg for 30 consecutive days.The normal group and the model group were given the same amount of normal saline by gavage.From the 21st day,except the normal group,the other groups were injected intraperitoneally with glyoxylic acid(10 mg/mL)to induce the kidney stone model for 10 d.On the 30th day,24 h urine of mice in each group was collected to detect Ox and Ca,blood was taken the next day to detect serum creatinine,urea nitrogen and Ca.Kidneys were dissected.Western blot was used to detect the expression levels of autophagy related proteins Beclin-1,LC3B,P62 and kidney injury molecule(KIM-1)in one kidney,and paraformaldehyde was used to fix the other kidney,and renal crystal deposition and renal histopathological changes were observed by HE staining.Results Compared with the model group,the water extract of Choerospondias axillaris could significantly reduce the level of urinary calcium(P<0.05),promote the excretion of urinary oxalic acid(P<0.05),significantly reduce the levels of serum creatinine and urea nitrogen(P<0.05),and had no significant effect on the level of serum Ca(P>0.05).The results of HE staining showed that the water extract of Choerospondias axillaris could significantly reduce the degree of renal tubular expansion,tubular degeneration and necrosis,fibrous tissue proliferation and crystal deposition in mice.The water extract of Choerospondias axillaris could significantly increase the expression levels of LC3BII/LC3BI(P<0.05),and significantly reduce the expression levels of KIM-1(P<0.05).Conclusion The water extract of Choerospondias axillaris can reduce renal injury and prevent the formation of renal calculi by activating autophagy.