脉血康胶囊减轻IgA肾病大鼠肾纤维化的作用和机制

Effects and mechanism of Maixuekang capsule on reducing renal fibrosis in IgA nephropathy rats

目的探讨脉血康胶囊对免疫球蛋白A肾病(IgAN)大鼠肾纤维化的改善及对Janus酪氨酸蛋白激酶2(JAK2)/信号转导与转录激活因子家族3(STAT3)信号通路的抑制作用。方法用600 mg·kg^(-1)改良的牛血清白蛋白(BSA)+脂多糖0.2 mL(LPS)+四氯化碳(CCl_(4))0.1 mL方案建立IgAN大鼠模型。将55只SD大鼠随机分为空白对照组(蒸馏水+0.9%NaCl)、模型组(IgAN模型)、阳性对照组(建模后灌胃给予替米沙坦片10 mg·kg^(-1))和低、中、高3个剂量实验组(建模后灌胃给予脉血康胶囊16、44和84 ATU·kg^(-1))。检测大鼠24 h尿蛋白;用全自动生化分析仪测定血肌酸酐(Scr)、血尿素氮(BUN)、谷丙转氨酶(GPT)、谷草转氨酶(GOT)水平;用苏木精-伊红(HE)染色法观察肾组织形态;用免疫荧光法检测IgA在大鼠肾中的沉积;用蛋白质印迹法检测肾组织中α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)、JAK2、STAT3、磷酸化STAT3(p-STAT3)蛋白的表达。结果空白对照组、模型组、阳性对照组与高剂量实验组的血Scr分别为(41.42±2.30)、(59.82±5.36)、(50.28±3.18)和(50.47±7.19)μmol·L^(-1),24 h尿蛋白分别为(2.35±1.21)、(11.62±3.45)、(5.01±4.36)和(8.14±4.19)mg·d^(-1),α-SMA蛋白相对表达水平分别为0.48±0.10、1.00±0.00、0.71±0.13和0.57±0.22,TGF-β1蛋白相对表达水平分别为0.54±0.93、1.00±0.00、0.73±0.33和0.51±0.31,JAK2蛋白相对表达水平分别为0.36±0.22、1.00±0.00、0.82±0.36和0.57±0.28,p-STAT3/STAT3分别为0.40±0.19、1.00±0.00、0.46±0.18和0.31±0.18。模型组的上述指标与空白对照组比较,在统计学上差异均有统计学意义(均P<0.05),阳性对照组和高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论脉血康胶囊可能通过抑制JAK2/STAT3信号通路的激活减轻IgAN大鼠肾纤维化水平,保护肾功能。

Objective To explore the effect of Maixuekang capsules on renal fibrosis and inhibition of Janus tyrosine protein kinase 2(JAK2)/signal transducer and activator of transcription family 3(STAT3)signal pathway in rats with immunoglobulin A nephropathy(IgAN).Methods The rat model of IgAN was established by modified 600 mg·kg^(-1)BSA+LPS 2 mL+CCL40.1 mL scheme,and the 24-hour urinary protein was detected.Fifty-five SD rats were randomly divided into 6 groups:Blank control group(distilled water+0.9%NaCl),model group(IgAN model),positive control group(10 mg·kg^(-1)telmisartan tablets after modeling),experimental-L,-M,-H groups(16,44,84 ATU·kg^(-1)Maixuekang capsules after modeling).The levels of serum creatinine(Scr),blood urea nitrogen(BUN),glutamic pyruvic transaminase(GPT)and glutamic oxalacetic transaminase(GOT)were determined by automatic biochemical analyzer.The morphology of kidney was observed by hematoxylin-eosin(HE)staining,and the deposition of IgA in rat kidney was detected by immunofluorescence.The expressions ofα-SMA,TGF-β1,JAK2,STAT3 and P-STAT3 in renal tissue were detected by Western blot.Results The blood Scr of blank control group,model group,positive control and experimental-H groups were(41.42±2.30),(59.82±5.36),(50.28±3.18)and(50.47±7.19)μmol·L^(-1),respectively;the 24-hour urinary protein were(2.35±1.21),(11.62±3.45),(5.01±4.36)and(8.14±4.19)mg·d^(-1),respectively;the relative expression ofα-SMA protein were 0.48±0.10,1.00±0.00,0.71±0.13 and 0.57±0.22,respectively;the relative expression of TGF-β1protein were 0.54±0.93,1.00±0.00,0.73±0.33 and 0.51±0.31,respectively;the relative expression of JAK2protein were 0.36±0.22,1.00±0.00,0.82±0.36 and 0.57±0.28,respectively;p-STAT3/STAT3 were0.40±0.19,1.00±0.00,0.46±0.18 and 0.31±0.18,respectively.Compared with the blank control group,the differences of above indexes in the model group were statistically significant(all P<0.05).Compared with the model group,the differences of above indexes in the positive control and experimental-H groups were statistically significant(all P<0.05).Conclusion Maixuekang capsules may reduce the level of renal fibrosis and protect renal function in rats with IgAN by inhibiting the activation of JAK2/STAT3 signal pathway.