Artemisinin ameliorated proteinuria in rats with adriamycin-induced nephropathy through regulating nephrin and podocin expressions

OBJECTIVE:To investigate the effects of artemisinin against proteinuria and glomerular filtration barrier damage in rats with adriamycin-induced nephropathy,and the potential mechanism underpinned the action.METHODS:Forty adriamycin rats were randomly divided into two groups with the ratio of 1︰3;the small-number group served as control group(n=10),and the rats in the large-number group were treated with adriamycin to induce nephropathy;then they were further randomly assigned into 3subgroups:benazepril group(n=10),artemisiningroup(n=10),and adriamycin group(n=10).The benazepril group and artemisinin group were treated with benazepril suspl(5.0 mg/kg daily)and artemisinin suspl(150 mg/kg daily)respectively after being modeled;those in the control group and adriamycin group were intragastrically administered an equivalent volume of distilled water every day.The treatment after model establishment lasted for a total of 4 weeks.The 24 h uric protein,blood biochemicals,renal pathological changes,renal ultrastrutural changes,Nephrin and Podocin proteins and gene expressions were measured by Coomassie brilliant blue assay,completely automatic biochemical analyzer,light microscope,electron microscopy,Western blot and reverse transcription polymerase chain reaction,respectively.RESULTS:The rats in adriamycin group showed a significant increase in 24 h uric protein excretion,serum total cholesterol(TC),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(Scr)and decrease in albumin(Alb)(P<0.05 or P<0.01).Compared with adriamycin group,artemisinin could reduce uric protein excretion,decrease the serum TC,TG elevation,increase the serum Alb level,up-regulate the expressions of Nephrin and Podocin(P<0.05 or P<0.01),but no statistical significance effects on the levels of BUN,Scr in artemisinin group(P>0.05).The renal pathological and ultrastrutural observation indicate that artemisinin could attenuate the severity of foot process effacement and fusion in the nephropathic rats.CONCLUSION:Artemisinin might have an effect on the nephropathy in rats caused by adriamycin,which may be at least partly correlated with attenu-ation of the severity of foot process effacement and fusion,up-regulation of the expressions of Nephrin and Podocin in the glomeruli in the rats.

Qufeng Tongluo Prescription(祛风通络方)Inhibits Mesangial Cell Proliferation and Promotes Apoptosis through Regulating Cell Cycle Progression

Objective： To study the effects and possible underlying mechanism of Qufeng Tongluo Prescription （祛风通络方, QFTL） on the regulation of mesangial cells （MCs） proliferation and apoptosis. Methods： The MCs used in this experiment have undergone five passages induced by lipopolysaccharide （LPS）. Changes in the proliferation, apoptosis, cell cycle regulatory proteins and mRNA expression levels of the MCs after administration of Benazepril or QFTL were measured by methyl thiazolyl tetrazolium （MTT） reduction assay, flow cytometry, Western blot and quantitative real-time polymerase chain reaction （qRT-PCR）, respectively. Results： The addition of Benazepril or QFTL serum inhibited LPS-induced MC proliferation after treatment for 24, 48 and 72 h, respectively （P〈0.05 or P〈0.01）. Moreover, the inhibitory effect is more significant in the QFTL group at 48 h （P〈0.05）. Compared with the control group, LPS-induced cell proliferation decreased the number of cells in G1 phase versus cells in S and G2/M phases, while the addition of QFTL and Benazepril serum increased the ratio of cells at G1 phase （P〈0.05 or P〈0.01） to cells at S phase （P〈0.01）, implicating the cell cycle inhibition effect exerted by QFTL. LPS decreased the level of MC apoptosis, compared with the control group （P〈0.05）, while QFTL and Benazepril serum increased the level of MC apoptosis （P〈0.01）. Moreover, the difference between the QFTL group and the Benazepril group was statistically significant （P〈0.01）. Compared with the control group, the protein and mRNA expression levels of cylinD1, cyclin dependent kinase 2 （CDK2） and p21 were significantly increased （P〈0.05 or P〈0.01）, p27 was decreased but with no statistical significance （P〉0.05）; After being treated with QFTL and Benazepril serum, the protein and mRNA expression levels of cylinD1, CDK2, p21 were decreased and p27 increased significantly （P〈0.05 or P〈0.01）; Compared with the Benazepril group, QFTL show better effects on protein and mRNA expression levels of cylinD1, CDK2 （P〈0.05 or P〈0.01） and p21 protein expression （P〈0.05）. Conclusion： QFTL inhibits MCs proliferation, promotes MCs apoptosis through an underlying mechanism of down-regulating the protein and mRNA expression levels of cylinD1, CDK2, p21 and up-regulation of the expression level of p27.