Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum album...Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum albumin (c-BSA) into tail vein for several times. model mice were randomly divided into MN group (equal amount of distilled water), Shenqi Zhilong Decoction low dose group (12 g crude drug/kg), Shenqi Zhilong Decoction high dose group (24 g crude drug/kg), and Tripterygium wilfordii polyglycoside tablet group (14 mg/ kg). Another 10 un-treatment mice were taken as control group (equal amount of distilled water). The drug was administered orally once a day for 4 weeks. After the last administration, 24 hours urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function, liver function, blood lipid and levels of IL-6, IL-4 and TNF-α in serum in each group;HE staining was used to observe the pathological changes of kidney. Immunohistochemical staining was used to observe the expression of IgG in kidney. The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method. The gene expression of Neph1, Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR. Results: Compared with model group, Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels (P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function (P<0.05 or P<0.01);has no significant effects on the levels of CRE, UREA and UA in renal function (P>0.05). Decreased the contents of IL-6, IL-4 and TNF-α in serum (P<0.05 or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice (P<0.05 or P<0.01);Significantly increased the expression levels of NephP1, Nephrin and Podocin mRNA in renal tissues (P<0.01). Conclusion: Shenqi Zhulong Decoction has a good therapeutic effect on MN mice, and the mechanism of action is related to regulate the expression of related genes of Nephrin-Podocin-Neph1 receptor complex for protecting the glomerular filtration barrier, and inhibite the activation of ERK/cPLA2 pathway for relieving damage of GEC and reduceing secretion of pro-inflammatory cytokines.展开更多
基金Fund Project:Heilongjiang Natural Science Foundation Project(No.LH2020H104)Heilongjiang Postdoctoral Fund(No.LBH-Z20033)。
文摘Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum albumin (c-BSA) into tail vein for several times. model mice were randomly divided into MN group (equal amount of distilled water), Shenqi Zhilong Decoction low dose group (12 g crude drug/kg), Shenqi Zhilong Decoction high dose group (24 g crude drug/kg), and Tripterygium wilfordii polyglycoside tablet group (14 mg/ kg). Another 10 un-treatment mice were taken as control group (equal amount of distilled water). The drug was administered orally once a day for 4 weeks. After the last administration, 24 hours urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function, liver function, blood lipid and levels of IL-6, IL-4 and TNF-α in serum in each group;HE staining was used to observe the pathological changes of kidney. Immunohistochemical staining was used to observe the expression of IgG in kidney. The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method. The gene expression of Neph1, Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR. Results: Compared with model group, Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels (P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function (P<0.05 or P<0.01);has no significant effects on the levels of CRE, UREA and UA in renal function (P>0.05). Decreased the contents of IL-6, IL-4 and TNF-α in serum (P<0.05 or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice (P<0.05 or P<0.01);Significantly increased the expression levels of NephP1, Nephrin and Podocin mRNA in renal tissues (P<0.01). Conclusion: Shenqi Zhulong Decoction has a good therapeutic effect on MN mice, and the mechanism of action is related to regulate the expression of related genes of Nephrin-Podocin-Neph1 receptor complex for protecting the glomerular filtration barrier, and inhibite the activation of ERK/cPLA2 pathway for relieving damage of GEC and reduceing secretion of pro-inflammatory cytokines.
