Yi Qi Qing Re Gao formula ameliorates puromycin aminonucleoside-induced nephrosis by suppressing inflammation and apoptosis

BMC Complementary and Alternative Medicine

Ping Li²  Xi Dong²  Haojun Zhang²  Liping Yang¹  Tingting Zhao²  Huijie Liu³  Yongli Zhan¹  Yumin Wen² 

[1] Department of Nephrology, Guang’anmen Hospital of China Academy of Traditional Chinese Medical Sciences, Beijing 100053, China;Beijing Key Lab for Immune-Mediated Inflammatory Diseases, Beijing 10029, China;Department of Nephrology, Shunyi Hospital of Traditional Chinese Medicine, Beijing 101300, China

关键词: YQQRG formula;    Tumor necrosis factor;    Puromycin aminonucleoside nephrosis;    Inflammation;    Apoptosis;   

Others  :  1210272
DOI  :  10.1186/s12906-015-0673-9

 received in 2014-11-22, accepted in 2015-05-13,  发布年份 2015

【 摘 要 】

Background

Yi Qi Qing Re Gao (YQQRG) formula is a traditional Chinese herbal medicine used to treat chronic nephritis. This study was designed to evaluate the underlying mechanism in the use of YQQRG formula to treat nephrosis induced by puromycin aminonucleoside (PAN).

Methods

Thirty-six male Wistar rats were randomly divided into 3 groups of 12 rats each: a sham group, a vehicle-treated PAN model group (PAN), and a group treated with YQQRG (PAN + YQQRG). The PAN model was established by a single intravenous injection of PAN at a dose of 40 mg/kg body weight; rats in the sham group received the same volume of saline. Twenty-four hour urinary protein was measured 0, 3, 5, 10, and 15 days after the injection. The rats were sacrificed on day 10 and day 15 and the serum lipid profile examined. The renal cortex of each rat was stained with periodic acid–Schiff reagent and the pathologic alterations and ultrastructural changes were examined by transmission electron microscopy. In situ cell apoptosis was detected by a terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end-labeling (TUNEL) assay. Transcriptive levels of inflammatory markers and molecules associated with apoptosis were detected by a real-time polymerase chain reaction and expression of proteins was examined by either immunohistochemistry or Western blot analysis.

Results

YQQRG significantly decreased urinary protein level, and lowered serum lipid level. YQQRG also attenuated histologic lesions in the rat kidneys. Activation of inflammatory markers was largely restored by the administration of YQQRG. TUNEL assay showed that YQQRG decreased the number of apoptotic cells. Both mRNA and protein levels of caspase-3 were significantly reduced in the group treated with YQQRG, whereas expression of the Bcl-2 protein increased in the YQQRG group.

Conclusions

YQQRG alleviated kidney injury in PAN-treated rats, possibly through anti-inflammatory and anti-apoptotic effects.

【 授权许可】

   
2015 Wen et al.; licensee BioMed Central.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

Fig. 7.

Fig. 8.

Fig. 9.

Fig. 10.

Fig. 11.

Fig. 12.

【 参考文献 】

• [1]Imig JD, Ryan MJ. Immune and inflammatory role in renal disease. Compr Physiol. 2013; 3(2):957-76.
• [2]Navarro-Gonzalez JF, Mora-Fernandez C, Muros de Fuentes M, Garcia-Perez J. Inflammatory molecules and pathways in the pathogenesis of diabetic nephropathy. Nat Rev Nephrol. 2011; 7(6):327-40.
• [3]Wang Y, Harris DC. Macrophages in renal disease. J Am Soc Nephrol. 2011; 22(1):21-7.
• [4]Sanz AB, Santamaria B, Ruiz-Ortega M, Egido J, Ortiz A. Mechanisms of renal apoptosis in health and disease. J Am Soc Nephrol. 2008; 19(9):1634-42.
• [5]Zoja C, Abbate M, Remuzzi G: Progression of renal injury toward interstitial inflammation and glomerular sclerosis is dependent on abnormal protein filtration. Nephrol Dial Transplant. 2015;30(5):706-712.
• [6]Zhan YDX. Treatment of 30 cases with chronic nephritis with replenishing qi, removing blood stasis, clearing heat and toxin methods. J Tradit Chin Med. 2003; 44:922-4.
• [7]Zhan YDX, Li X, Li S, Rao X. Renoprotective effect of Yiqiqingre extract in Adriamycin nephritic rats. Chinese J Integr Tradit Western Nephrol. 2003; 4:135-8.
• [8]Zhan Y, Yang L, Wen Y, Liu H, Zhang H, Zhu B et al.. Yi qi qing re gao attenuates podocyte injury and inhibits vascular endothelial growth factor overexpression in puromycin aminonucleoside rat model. Evidence-Based Complement Altern Med: eCAM. 2014; 2014:375986.
• [9]Kruger NJ. The Bradford method for protein quantitation. Methods Mol Biol. 1994; 32:9-15.
• [10]Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008; 3:1101-8.
• [11]Hosoyamada M, Yan K, Nishibori Y, Takiue Y, Kudo A, Kawakami H et al.. Nephrin and podocin expression around the onset of puromycin aminonucleoside nephrosis. J Pharmacol Sci. 2005; 97:234-41.
• [12]Gomez-Chiarri M, Ortiz A, Lerma JL, Lopez-Armada MJ, Mampaso F, Gonzalez E et al.. Involvement of tumor necrosis factor and platelet-activating factor in the pathogenesis of experimental nephrosis in rats. Lab Invest. 1994; 70(4):449-59.
• [13]Mampaso FM, Egido J, Martinez-Montero JC, Bricio T, Gonzalez E, Cobo ME et al.. Interstitial mononuclear cell infiltrates in experimental nephrosis: effect of PAF antagonist. Nephrol Dial Transplant. 1989; 4(12):1037-44.
• [14]Ou ZL, Natori Y, Natori Y. Transient and sequential expression of chemokine mRNA in glomeruli in puromycin aminonucleoside nephrosis. Nephron. 2000; 85(3):254-7.
• [15]Caulfield JP, Reid JJ, Farquhar MG. Alterations of the glomerular epithelium in acute aminonucleoside nephrosis. Evidence for formation of occluding junctions and epithelial cell detachment. Lab Invest. 1976; 34(1):43-59.
• [16]Gwinner W, Landmesser U, Brandes RP, Kubat B, Plasger J, Eberhard O et al.. Reactive oxygen species and antioxidant defense in puromycin aminonucleoside glomerulopathy. J Am Soc Nephrol. 1997; 8(11):1722-31.
• [17]Rincon J, Romero M, Viera N, Pedreanez A, Mosquera J. Increased oxidative stress and apoptosis in acute puromycin aminonucleoside nephrosis. Int J Exp Pathol. 2004; 85(1):25-33.
• [18]Ni Z, Vaziri ND. Downregulation of nitric oxide synthase in nephrotic syndrome: role of proteinuria. Biochim Biophys Acta. 2003; 1638(2):129-37.
• [19]Walker LM, Shah SV, Mayeux PR. Lack of a role for inducible nitric oxide synthase in an experimental model of nephrotic syndrome. Biochem Pharmacol. 2000; 60(1):137-43.
• [20]Fernandez L, Romero M, Soto H, Mosquera J. Increased apoptosis in acute puromycin aminonucleoside nephrosis. Exp Nephrol. 2001; 9(2):99-108.
• [21]Tharaux PL, Huber TB. How many ways can a podocyte die? Semin Nephrol. 2012; 32(4):394-404.
• [22]Pippin JW, Brinkkoetter PT, Cormack-Aboud FC, Durvasula RV, Hauser PV, Kowalewska J et al.. Inducible rodent models of acquired podocyte diseases. Am J Physiol Renal Physiol. 2009; 296(2):F213-29.
• [23]Ohse T, Inagi R, Tanaka T, Ota T, Miyata T, Kojima I et al.. Albumin induces endoplasmic reticulum stress and apoptosis in renal proximal tubular cells. Kidney Int. 2006; 70(8):1447-55.
• [24]Muroya Y, Ito O, Rong R, Takashima K, Ito D, Cao P et al.. Disorder of fatty acid metabolism in the kidney of PAN-induced nephrotic rats. Am J Physiol Renal Physiol. 2012; 303(7):F1070-9.
• [25]Lee HS. Mechanisms and consequences of hypertriglyceridemia and cellular lipid accumulation in chronic kidney disease and metabolic syndrome. Histol Histopathol. 2011; 26(12):1599-610.
• [26]Brentnall M, Rodriguez-Menocal L, De Guevara RL, Cepero E, Boise LH. Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol. 2013; 14:32. BioMed Central Full Text
• [27]Soriano ME, Scorrano L. The interplay between BCL-2 family proteins and mitochondrial morphology in the regulation of apoptosis. Adv Exp Med Biol. 2010; 687:97-114.
• [28]Riedl SJ, Shi Y. Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol. 2004; 5(11):897-907.
• [29]Gui D, Huang J, Guo Y, Chen J, Chen Y, Xiao W et al.. Astragaloside IV ameliorates renal injury in streptozotocin-induced diabetic rats through inhibiting NF-kappaB-mediated inflammatory genes expression. Cytokine. 2013; 61(3):970-7.
• [30]He CL, Yi PF, Fan QJ, Shen HQ, Jiang XL, Qin QQ et al.. Xiang-Qi-Tang and its active components exhibit anti-inflammatory and anticoagulant properties by inhibiting MAPK and NF-kappaB signaling pathways in LPS-treated rat cardiac microvascular endothelial cells. Immunopharmacol Immunotoxicol. 2013; 35(2):215-124.
• [31]Tzeng TF, Liou SS, Chang CJ, Liu IM. The ethanol extract of Lonicera japonica (Japanese honeysuckle) attenuates diabetic nephropathy by inhibiting p-38 MAPK activity in streptozotocin-induced diabetic rats. Planta Med. 2014; 80(2–3):121-9.
• [32]Muluye RA, Bian Y, Alemu PN. Anti-inflammatory and Antimicrobial Effects of Heat-Clearing Chinese Herbs: A Current Review. J Tradit Complement Med. 2014; 4(2):93-8.
• [33]Pan X, Cao X, Li N, Xu Y, Wu Q, Bai J et al.. Forsythin inhibits lipopolysaccharide-induced inflammation by suppressing JAK-STAT and p38 MAPK signalings and ROS production. Inflamm Res. 2014; 63(7):597-608.
• [34]Yang EJ, Lee JS, Yun CY, Kim JH, Kim JS, Kim DH et al.. Inhibitory effects of Duchesnea chrysantha extract on ovalbumin-induced lung inflammation in a mouse model of asthma. J Ethnopharmacol. 2008; 118(1):102-7.
• [35]Zhu H, Liang QH, Xiong XG, Chen J, Wu D, Wang Y et al.. Anti-Inflammatory Effects of the Bioactive Compound Ferulic Acid Contained in Oldenlandia diffusa on Collagen-Induced Arthritis in Rats. Evidence-Based Complement Altern Med: eCAM. 2014; 2014:573801.
• [36]Jia Y, Zuo D, Li Z, Liu H, Dai Z, Cai J et al.. Astragaloside IV inhibits doxorubicin-induced cardiomyocyte apoptosis mediated by mitochondrial apoptotic pathway via activating the PI3K/Akt pathway. Chem Pharm Bull. 2014; 62(1):45-53.
• [37]Kim MH, Kim SH, Yang WM. Beneficial effects of Astragaloside IV for hair loss via inhibition of Fas/Fas L-mediated apoptotic signaling. PLoS One. 2014; 9(3):e92984.
• [38]Wang Q, Shao X, Xu W, Qi C, Gu L, Ni Z et al.. Astragalosides IV inhibits high glucose-induced cell apoptosis through HGF activation in cultured human tubular epithelial cells. Ren Fail. 2014; 36(3):400-6.
• [39]Li Y, Shi W, Li Y, Zhou Y, Hu X, Song C et al.. Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury. Environ Toxicol Pharmacol. 2008; 26(1):13-21.
• [40]Domitrovic R, Cvijanovic O, Pugel EP, Zagorac GB, Mahmutefendic H, Skoda M. Luteolin ameliorates cisplatin-induced nephrotoxicity in mice through inhibition of platinum accumulation, inflammation and apoptosis in the kidney. Toxicology. 2013; 310:115-23.
• [41]Liu XH, Pan LL, Gong QH, Zhu YZ. Antiapoptotic effect of novel compound from Herba leonuri - leonurine (SCM-198): a mechanism through inhibition of mitochondria dysfunction in H9c2 cells. Curr Pharm Biotechnol. 2010; 11(8):895-905.