Ameliorative effects of arctiin from Arctium lappa on experimental glomerulonephritis in rats.
Membranous glomerulonephritis (MGN) remains the most common cause of adult-onset nephrotic syndrome in the world and up to 40% of untreated patients will progress to end-stage renal disease. Although the treatment of MGN with immunosuppressants or steroid hormones can attenuate the deterioration of renal function, numerous treatment-related complications have also been established. In this study, the ameliorative effects of arctiin, a natural compound isolated from the fruits of Arctium lappa, on rat glomerulonephritis induced by cationic bovine serum albumin (cBSA) were determined. After oral administration of arctiin (30, 60, 120mg/kgd) for three weeks, the levels of serum creatinine (Ser) and blood urea nitrogen (BUN) and 24-h urine protein content markedly decreased, while endogenous creatinine clearance rate (ECcr) significantly increased. The parameters of renal lesion, hypercellularity, infiltration of polymorphonuclear leukocyte (PMN), fibrinoid necrosis, focal and segmental proliferation and interstitial infiltration, were reversed. in addition, we observed that aretiin evidently reduced the levels of malondialdehyde (MDA) and pro-inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor (TNF-[alpha]), suppressed nuclear factor-kB [p.sup.65] (NF-[kappa]B) DNA binding activity, and enhanced superoxide dismutase (SOD) activity. These findings suggest that the ameliorative effects of arctiin on glomerulonephritis is carried out mainly by suppression of NF-[kappa]B activation and nuclear translocation and the decreases in the levels of these pro-inflammatory cytokines, while SOD is involved in the inhibitory pathway of NF-[kappa]B activation. Arctiin has favorable potency for the development of an inhibitory agent of NF-[kappa]B and further application to clinical treatment of glomerulonephritis, though clinical studies are required.
[c] 2009 Elsevier GmbH. Ali rights reserved.
Keywords: Arctiin; Arctium lappa: Glomerulonephritis; Nuclear factor-KB; Pro-inflammatory cytokine; Superoxide dismutase
Membranous glomerulonephritis (MGN), a prototype of an immune-mediated glomerular disease, is characterized by abundant, nonselective proteinuria and a highly individualed course and prognosis (Glassock et. al. 1996). MGN remains the most common cause of adult-onset nephrotic syndrome in the world (Cattran 2001). Studies on the natural history of the disease show that up to 40% of untreated patients will progress to end-stage renal disease (Schieppati et al. 1993; Ponticelli et al. 1995; Stirling et al. 1998). Treatments with both steroid hormones and immunosuppressive agents are so unsatisfactory that side effects such as bone marrow depression, infection, cancer, and so on, occur frequently and high relapse rate is also observed (Peggy et al. 2004). Therefore, the search for drugs with fewer side effects is important for preventing and delaying the development and progression of MGN.
Chinese medicines are therapeutic agents based on natural plants and have been used to treat various human diseases under the guidance of the theory of Chinese medical sciences in China for centuries. People are becoming increasingly interested in Chinese medicines for their low toxicity and good therapeutic performance. The search for new drugs capable of improving renal survival with fewer side effects has gained momentum over the years, resulting in numerous reports on significant activities of Chinese medicines (Yin et al. 2005: Bao et al. 2004: Mei et al. 2004; Huang and Liu 2008). More attention is focused on bioactive components from Chinese medicines in recent years. Although plant extracts constitute potential candidates, they often contain highly complex mixtures of many different compounds (Zhang et al. 2008a-c), which are often incompletely known. The aim of this study is to find a purified compound which can improve glomerulonephritis.
Fructus Arctii, the dried fruits of Arctium lappa L., is one of the most popular Chinese medicines and is officially listed in the Chinese Committee of National Pharmacopoeia (2005). It has been widely used for dispelling pathogenic wind-heat, promoting eruption, relieving sore throat, removing toxic substances and subduing swelling. Arctiin (chemical structure seen in Fig. 1), the most lignan compound in Fructus Arctii, possesses many kinds of bioactivities and a number of important pharmacological properties proved by previous studies, including demutagenic (Shinohara et al. 1988), cytoxic (Moritani et al. 1996), anti-proliferative (Moritani et al. 1996), enhancement of immunological (Yan and Li 1993), anti-inflammatory (Yan and Li 1993), anti-carcinogenesis (Hirosea et al. 2000), platelet activating factor antagonist (Iwakami et al. 1992) and calcium antagonist (Ichikawa et al. 1986) activities. Arctiin also has protective effects on 2-amino-l-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP)-induced carcinogenesis particularly in the mammary gland in the promotion period by testing on Sprague-Dawley rats (Hirosea et al. 2000) and remarkable anti-tumor-promoting effects on carcinogenesis test of mouse skin tumors induced by 7,12-dimethylbenzanthracene (Takasaki et al. 2000). In addition, arctiin could significantly induce cell detachment and reduce the cell numbers through up-regulation of MUC-1 mRNA and protein in PC-3 cells (Huang et al. 2004), evidently ameliorate the blood rheology abnormality and enhance anti-coagulation effects in experimental rats with the blood stasis (Zheng et al. 2006). Nevertheless, to our knowledge, there have been no reports yet on the effects of arctiin in rat glomerulonephritis induced by cationic bovine serum albumin (cBSA).
[FIGURE 1 OMITTED]
In the present study, the glomerulonephritis rat model induced by cBSA was employed, and the parameters of renal function impairment and glomerular lesion were measured, including scrum creatinine (Scr), blood urea nitrogen (BUN), and endogenous creatinine clearance rate (ECcr), 24-h urine protein, hypercellularity, infiltration of polymorphonuclear leukocyte (PMN), fibrinoid necrosis, focal and segmental proliferation, and interstitial infiltration. Furthermore, pro-inflammatory cytokines including IL-6 and [TNF-[alpha]], superoxide dismutase (SOD), malondialdehyde (MDA), and nuclear factor-kB (NF-[kappa]B) were also investigated. This study was designed to assess whether or not arctiin possesses ameliorative effects on glomerulonephritis and its possible action pathway.
Materials and methods
Arctiin was isolated and purified from the fruits of Arctium lappa L. according to a previous report (Wang et al. 2005) by Professor Lian-Na Sun, a phytochemist from the Department of Pharmacognosy, School of Pharmacy, Second Military Medical University (Shanghai, China), and dissolved in distilled water prior to administration. (Molecular formula: [[C.sub.27][H.sub.34][O.sub.11]]; Molecular weight: 534.54; Purity: 98.2%; Appearance: colorless crystalline powder).
Bovine serum albumin (BSA, Sigma), purchased from Sino-American Biotechnology Company, Shanghai), was used as substrate to make charge-modified cationic bovine serum albumin (cBSA). The method described by Hoare and Koshland (1967) was performed.
Animals and grouping
Fifty Sprague-Dawley male rats were obtained from the Experimental Animal Center of the Second Military Medical University (Shanghai, China), with an initial body weight of 130g + or - 20g. They were housed in a regulated environment (22 + or - 2[degrees]C), with a 12-h dark and 12-h light cycle (08:00-20:00, light). Food and water were given ad libitum, except for the duration of the experimental period. All animal treatments were strictly in accordance with international ethical guidelines and the National Institutes of Health Guide concerning the Care and Use of Laboratory Animals, and the experiments were carried out with the approval of the Committee of Experimental Animal Administration of the University.
After the 7-day acclimation period, animals were randomly divided into 5 groups (n = 10): one normal group, one control group, and three groups treated with arctiin (30, 60, and 120 mg [kg.sup.-1] per day).
The experimental design was based on cBSA-induced glomerulonephritis according to the method reported by Border et al. (1982). Briefly, the rats in the control group and three arctiin-treated groups were received a primary subcutaneous immunization with cBSA (1.0 mg in 0.5 ml 0.01 M phosphate buffered saline plus 0.5 ml incomplete freund's adjuvant). One week later, cBSA (2.5 mg in 1 ml 0.01 M PBS) was injected through the caudal vein every other day for 3 weeks to induce glomerulonephritis. Rats in the normal group were injected with isometrical normal saline through the caudal vein every other day from week 2 to week 4. At the same time, three treatment groups of animals were orally administered 30, 60, 120 mg/kg [d.sup.-1] of arctiin in a volume of 10 ml by intubation, respectively, for 3 weeks. Other groups of animals were orally administered the same volume of distilled water, and they were run concurrently with arctiin-treated groups.
The urine samples were collected in week 0, week 2, week 3, and week 4, respectively, after the administration of arctiin. At the end of the experiment, all animals were anesthetized with 40 mg/kg of sodium pentobarbital and sacrificed at 1 h after the last administration. Blood samples were collected to measure BUN, Scr, ECcr, IL-6, and TNF-[alpha]. The right renal cortex of animals was homogenized in Tris-HCl buffer (0.05 mol/I Tris-HCl, 1.15% KC1, [p.sup.H 7.4]) using a homogeniser. The homogenate was centrifuged at 15,000 x g (4[degrees]C) for 30 min, and the supernatant was utilized for the measurement of NF-[kappa]B, MDA, and SOD. The left kidneys were used for histological examination.
Assay for 24-h urine protein quantification
The rats were placed in metabolic cages for 24-h urine collection in week 0, week 2, week 3, and week 4, Each rat remained on its ambient diet with an unrestricted fluid intake. 24 h later, the volume of urine was measured. Individually collected urine was centrifuged at 4000 X g for 5 min. The supernatant was collected and urine proteins were quantified by the Coomassie brilliant blue colorimetric method.
Assessment of kidney function
At the end of the experimental period, Animals were sacrificed and blood (6 ml) obtained from each rat was centrifuged at 5000 X g at room temperature for 10 min. The serum was collected and stored at -70 [degrees]C for the detection of serum creatinine (Scr) and blood urea nitrogen (BUN). The examination of the biochemical parameters was performed by an automatic biochemistry analyzer (Hitachi 7180, Hitachi, Japan).
Endogenous creatinine clearance rate (ECcr) was calculated from the values of Scr, urine creatinine (Ucr) and total 24-h urine volume, viz. Ucr /Scr X 24 h urine volume (ml/min).
Determination of IL-6 and TNF-[alpha]
The serum levels of IL-6 and TNF-[alpha] were determined by enzyme-linked immunosorbent assay (ELISA) using commercially available kits (R&D Systems). All samples and provided standards were analyzed in duplicate. A standard curve was constructed using standards provided in the kits, and the cytokine concentrations were determined from the standard curves using linear regression analysis.
Assessment of NF-[kappa]B DNA binding activity
NF-[kappa]B DNA binding in nuclear extracts was determined using a commercially available NF-[kappa]B p65 ELISA kit according to the manufacturer's instructions (TransAM ELISA kit, Active Motif). Nuclear protein was obtained from tissue lysates, prepared from the fresh renal cortical tissues of the rats. The amount of bound NF-[kappa]B p65 was detected by adding IRDyeconjugated, anti-rabbit secondary antibody. Optical density (OD) value was read at 450 nm using colorimetric detection in an ELISA plate reader. All results are expressed as averages of the two duplicate samples assayed after subtracting the average of the two blank values (Renard et al. 2001).
Measurement of MDA and SOD
The determination of superoxide dismutase (SOD) activity was based on the inhibition of pyrogallol autooxidation (Misra and Fridovich 1972). The concentration of malondialdehyde (MDA) was determined by the reaction with thiobarbituric acid (Ohkawa et al. 1979).
Histological examination was in accordance with the method reported by Mirshafiey et al. (Mirshafiey et al. 2000). Renal tissues were fixed with 10% buffered formalin for 3 days, embedded in paraffin, and their 3-[ micro]m sections were stained with hematoxilin-esosin (HE) and periodic acid-Schiff (PAS). Kidney specimens were processed under light microscope. Five parameters, including hypercellularity, infiltration of polymorphonuclear leukocyte (PMN), fibrinoid necrosis, focal and segmental proliferation, and interstitial infiltration, were used to blindly estimate the severity and extent of glomerular lesion. These indices were evaluated by a semi-quantitative method of renal histology using a grading scale of 0-3 (0, negative; 1, mild; 2, moderate; 3, severe).
The results were analyzed using a SPSS 11.0 statistical package. The data for multiple comparisons were performed by one-way ANOVA followed by Dunnett t-test. A value of p <0.05 was considered statistically significant and all results are presented as the mean [+ or -] s.e.m.
Effects on 24-h urine protein content in glomerulonephritis rats
As shown in Table 1, there was no difference in initial 24-h urine protein content in different groups of rats. However, injection of cBSA resulted in a significant increase (p <0.01) at week 2, week 3 and week 4. Co-administration of arctiin (60, 120 mg/kg) markedly decreased the levels of urine protein at ail time points measured (p <0.05 or p <0.01), though the lower dose of arctiin (30 mg/kg) reduced urine protein levels only at week 4 (p <0.05)
Table 1. Effect of artiin on 24-h urine protein content in glomerulonephritis rats Group Animal (N) Dose (mg/kg [d.sup.-1]) Week 0 (mg) Normal 10 Distilled water 3.91[+ or -]0.13 Control 10 Distilled water 4.01[++or--0.12 Arctiin 10 30 4.00[+ or -]0.13 Arctiin 10 60 3.92[+ or -]0.11 Arctiin 10 120 3.88[+ or -]0.10 Group Week 2 (mg) Normal 4.12[+ or -]0.12 Control 95.44[+ or -]3.10 [DELTA][DELTA] Arctiin 88.85[+ or -]3.25 Arctiin 81.35[+ or -]2.67 ** Arctiin 72.38[+ or -]3.37 ** Group Week 3 (mg) Normal 4.27[+ or -] Control 115.45[+ or -]5.12 [DELTA][DELTA] Arctiin 101.32[+ or -]4.96 Arctiin 94.25[+ or -]3.67 * Arctiin 85.52[+ or -]3.40 ** Group Week 4 (mg) Normal 4.18[+ or -] Control 142.23[+ or -]4.70 [DELTA][DELTA] Arctiin 117.86[+ or -]4.51 * Arctiin 93.41[+ or -]3.93 ** Arctiin 88.69[+ or -]3.06 ** [DELTA][DELTA] P<0.01 compared with the normal group, * p<0.05, ** p<0.01 compared with the control group. Delta are expressed as the mean [+ or -]s.e.m for ten samples per treatment group.
Effects on renal function in glomerulonephritis rats
Table 2 indicates that the injection of cBSA for 4 weeks caused significant deterioration of renal function in rats. The levels of serum BUN and Scr evidently increased (<0.0l), while the ECcr level decreased (<0.01), compared with those in the control group. As expected, the renal function was markedly ameliorated in the groups of animals treated with arcttin (30, 60, 120mg/kg) for 3 weeks, compared with that of the control group (<0.05 or <0.01).
Table 2. Effects of arctiin on renal function in glomerulonephritis rats. Group Animal Dose (mg/kg BUN (mmol/1) (N) [d.sup.-1]) Normal 10 Distilled water 7.64[+ or -]0.49 Control 10 Dustilled water 10.81[+ or -]0.55[DELTA][DELTA] Arctiin 10 30 8.41[+ or -]0.59* Arctiin 10 60 8.03[+ or -]0.50** Arctiin 10 120 7.67[+ or -]0.39** Group Scr ([mu]mol/1) Normal 50.75[+ or -]1.71 Control 74.84[+ or -]3.10 [DELTA][DELTA] Arctiin 65.46[+ or -]1.47 * Arctiin 63.16[+ or -]2.02 ** Arctiin 56.49[+ or -]2.05 ** Group Eccr (ml/min) Normal 2.08[+ or -]0.08 Control 1.15[+ or -]0.07 [DELTA][DELTA] Arctiin 1.28[+ or -]0.07 Arctiin 1.42[+ or -]0.06 ** Arctiin 1.74[+ or -]0.05 ** Table 2 showed the effects of arctiin on renal function in glomerulonephritis rats after injection of cBSA for 4 weeks. [DELTA][DELTA] P<0.01 compared with the normal group, * p<0.05, ** p<0.01 compared with the control group. Data are expressed as the mean [+ or- ]s.e.m for ten samples per treatment group.
Effects on serum 1L-6 and TNF-[alpha] concentration in glomerulonephritis rats
It is showed in Fig. 2 that the serum levels of IL-6 and TNF-[alpha] were significantly higher in the control group than in the normal group (<0.01). Nevertheless, co-administration of arctim (30, 60, 120mg/kg) for 3 weeks dramatically reduced their serum levels in a dose-dependent manner compared with the control group (<0.01).
[FIGURE 2 OMITTED]
Effects on NF-kd DNA binding activity from renal tissues
The effects of arctiin treatment on NF-[kappa]B DNA binding activity from renal tissues are shown in Fig. 3. There was a significant increase in the NF-[kappa]B DNA binding activity from the renal tissues of rats treated with cBSA compared with the normal group of animals (<0.01). After arctiin (30, 60, 120mg/kg) treatment for 3 weeks, the NF-[kappa]B DNA binding activity dramatically decreased compared with the control group (<0.01), though it was not restored to the normal level.
Effects on kidney MDA level and SOD activity in glomerulonephritis rats
[FIGURE 3 OMITTED]
The renal tissue MDA level in the control group of animals was higher than that in the normal group of animals, while the SOD activity decreased (<0.0l, Fig. 4). Arctiin treatment (30, 60, 120 mg/kg) could significantly reduce the MDA level and increase the SOD activity when compared with the control group (<0.01).
[FIGURE 4 OMITTED]
Light microscopic examination of renal tissues revealed the severity and extent of glomerular lesion in the control group of animals (Table 3). Five parameters, including hypercellularity, glomerular infiltration of PMN, fibrinoid necrosis, focal and segmental proliferation, and interstitial infiltration, were markedly ameliorated in arctiin-treated animals compared with the control group of animals.
Table 3. Effects of arctiin on renal lesions in glomerulonephritis rats. Group Animal Dose (mg/kg Hypercellularity (N) [d.sup.-1]) Normal 10 Distilled 0 water Control 10 Distilled 2.40[+ or -]0.22 [DELTA][DELTA] water Arctiin 10 30 1.60[+ or -]0.31 Arctiin 10 60 1.30[+ or -]0.30 ** Arctiin 10 120 0.80[+ or -]0.25 ** Group PMN infiltration Fibrinoid necrosis Normal 0 0 Control 2.10[+ or -]0.23 [DELTA][DELTA] 1.30[+ or -]0.15 [DELTA][DELTA] Arctiin 1.10[+ or -]0.31 * 0.60[+ or -]0.22 ** Arctiin 1.00[+ or -]0.26 ** 0.40[+ or -]0.16 ** Arctiin 0.60[+ or -]0.22 ** 0.10[+ or -]0.10 ** Group focal and segmental Interstitial infiltration proliferation Normal 0 0 Control 0.70[+ or -]0.15 [DELTA][DELTA] 1.20[+ or -]0.20 [DELTA][DELTA] Arctiin 0.40[+ or -]0.16 0.40[+ or -]0.16 ** Arctiin 0.2[+ or -]0.13 * 0.30[+ or -]0.15 ** Arctiin 0 ** 0 ** [DELTA][DELTA] P<0.01 compared with the normal group, * p<0.05, ** p<0.01 compared with the control group. Data are expressed as the mean [+ or -] s.e.m for ten samples per treatment group.
Membranous glomerulonephritis (MGN) has been defined as granular subepithelial deposition of immune complexes along the glomerular basement membrane (GBM) (Tateno et al. 1997). Several animal models of immune complex glomerular injury have been developed to investigate the immunological mechanisms of human glomerulonephritis. Cationic bovine serum albumin (cBSA) glomerulonephritis, one of these immune complex glomerulonephritis models, is known to be induced by the administration of cBSA with incomplete freund's adjuvant in different kinds of experimental animals (Tateno et al. 1997), and is characterized by proliferative glomerulonephritis with immune deposit of glomeruli and leukocyte infiltration and low serum complement levels (Hunsicker et al. 1979; Ren et al. 1991). The pathogenesis of the model is that cBSA with positive charges is combined onto GBM with negative charges and forms immune complexes in glomeruli. These immune complexes may activate the complement system and result in glomerular imune injury that induces glomerulonephritis and finally progresses to renal function impairment (Couser and Salant 1980; Couser 1985; Paccarld 1989). This animal model exhibits great similarity to human MGN disease (Chen et al. 2004).
In the current study, cBSA-induced glomerulonephritis rat model revealed severe renal function impairment, which was manifested by the reduction in glomerular filtration rate, appearing as increases in Scr and BUN and a decrease in ECcr. There were significant differences in these parameters between the control group of rats and the normal group of animals. In contrast, the treatment with arctiin for 3 weeks reversed these indices.
The immune process causes generation of terminal complement complexes, which are essential for the development of proteinuria (Rother et al. 1991). Progression of MGN is correlated with the duration and degree of proteinuria (Honkanen et al. 1994; Cattran et al. 1997). We observed in this study that 24-h urine protein displayed a marked increase in model rats, while co-administration of arctiin evidently decreased it.
Renal function impairment and proteinuria are correlated well with the renal lesion, which can be detected via histological examination. Severe glomerular lesion was revealed in the control group of rats induced by cBSA. Five parameters, including hypercellularity, glomerular infiltration of PMN, fibrinoid necrosis, focal and segmental proliferation, and interstitial infiltration, evidently deteriorated compared with the normal rats. However, arctiin evidently improved these parameters. These results exhibited the protective effects of arctiin on glomerular lesion and renal function impairment in glomerulonephritis.
Pathophysiological studies of glomerulonephritis have demonstrated the pathogenic significance of pro-inflammatory cytokines. Renal insult induces the production of pro-inflammatory cytokines such as TNF-[alpha], IL-6, IL-1, and IL-8 (Atkins 2002), leading to leucocyte recruitment activation and subsequently haematuria, proteinuria and renal damage. However, the expression of these pro-inflammatory cytokines is transcriptionally regulated by nuclear factor-kB (NF-[.sub k]B), which plays a central role in the expression of a large number of genes involved in the inflammatory and immune responses (Baeuerle and Henkel 1994). Generally, NF-[.sub k]B is activated in a variety of cells in response to certain stimuli such as pro-inflammatory cytokines, oxidants, and viruses (Guijarro and Egido 2001; Wajant 2004; Xing et al. 2008), while activation of NF-[kappa]B leads to phosphorylation, ubiquitination, and then degradation of IkB-[alpha], leaving lipopolysaccharide (NF-[kappa]B) free to translocate to the nucleus from cytoplasm. Binding of NF-[kappa]B to the promoter region of certain genes results in an increase in their transcription (Atkins 2002). Suppression of NF-[kappa]B activation by certain inhibitors such as pyrrolidine dithiocarbamate or prednisolone could down-regulate the gene expression of pro-inflammatory cytokines, and then block the development of glomerulonephritis (Sakurai et al. 1996). In the current study, NF-[kappa]B [p.sup.65] was activated by repetitive administration of cBSA, accompanied by significant increases in the levels of IL-6 and TNF-[alpha]. In contrast, arctiin reversed these changes, indicating that arctiin may be a potential inhibitor of NF-[kappa]B.
A relationship between glomerulonephritis and oxygen free radicals has been confirmed in many experimental models (Walker and Shah 1987, 1988; Kumar et al. 2000). It has been demonstrated that antioxidant enzymes, such as superoxide dismutase (SOD), catalase, GSH-Px, and GSH-Rd, represent one protection against oxidative tissue-damage (Halliwell and Gutteridge 1990). SOD is an exceedingly effective defense enzyme that catalyzes the dismutalion of superoxide to hydrogen peroxide, which can then be further detoxified through the action of catalases and peroxidases. A lower level of SOD activity in patients with glomerulonephritis results in a decrease in the scavenger reaction of superoxide and causes renal tissue to become more vulnerable to oxidative stress (Kashem et al. 1996). MDA, a major reactive aldehyde that appears during the peroxidation of biological membrane polyunsaturated fatty acid, is widely used as a marker of free radical mediated lipid peroxidation injury (Vaca et al. 1988). The production of MDA parallels lipid peroxidation, but lipid peroxidation may conduce to the generation of MDA. The increase in MDA level in renal tissue suggests enhanced peroxidation leading to tissue damage and failure of the antioxidant defense mechanisms to prevent the formation of excessive free radicals (Naik 2003). The administration of SOD can result in a reduction in renal MDA production in glomerulonephritis animals (Rehan et al. 1984; Adachi et al. 1986; Birtwistle et al. 1989).
In the present study, SOD activity of renal tissues in rats significantly decreased and MDA level increased in response to cBSA treatment alone compared with normal group of rats. On the contrary, the treatment with arctiin reversed these changes. The administration of arctiin to cBSA-induced glomerulonephriti rats for 3 weeks markedly increased renal SOD activity, followed by a decrease in MDA level, in comparison to the control group of rats. These results verified that arctiin played a protective role in the reduction in oxidative stress. Because many investigations have demonstrated that the nuclear translocation can be inhibited by antioxidants such as pyrrolidine dithiocarbamate (PDTC) and resveratrol and oxygen free radicals are involved in the NF-[kappa]B activation pathway (Toledano and Leonard 1991; Schreck et al. 1992; Schreck and Baeuerle 1994; Kundu and Surh 2004), we speculate that the inhibition of NF-[kappa]B activation is well related to an increase in SOD activity by arctiin.
Nevertheless, arctiin is not directly involved in the regulation of NF-[kappa]B activation. After oral administration, arctiin is transformed into arctigenin, an aglycone of arctiin, in gastrointestinal tract and liver, and then exerts various activities in vivo (Nose et al. 1992). Arctigenin, but not arctiin, is detected in the scrum (Nose et al. 1993). On the other hand, arctigenin possesses potent antioxidant and anti-inflammatory activities, which can inhibit in vitro lipopolysaccharide-induccd nuclear NF-[kappa]B activation, accompanied by a decrease in TNF-[alpha] production (Cho et al. 2002, 2004). Thus, we presumed that arctiin, after transformation into arctigenin in vivo, suppressed the nuclear NF-[kappa]B activation, and then increased SOD activity, which may constitute anti-inflammatory effects of arctiin on glomerulonephritis.
In summary, arctiin, isolated from the fruits of Arctium lappa, has ameliorative effects on cBSA-induced glomerulonephritis in rats. It could evidently improve renal function, protect against glomerular lesion, decrease the levels of IL-6, TNF-[alpha], and MDA, increase the activity of SOD, and inhibit the NF-[kappa]B [p.sup.65] DNA binding activity. One of the probable mechanisms for these results is that arctiin, after transformation into arctigenin, suppresses NF-[kappa]B activation and nuclear translocation, followed by the decreases in the levels of these pro-inflammatory cytokines, while SOD is involved in the inhibitory pathway of NF-[kappa]B activation. Our investigation suggests that arctiin has favorable potency for the development of an inhibitory agent of NF-[kappa]B and further application to clinical treatment of glomerulonephritis, though clinical studies are required.
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Jian-Guo Wu (a), (1), Jin-Zhong Wu (b), (1), Lian-Na Sun (a), Ting Han (a), Jian Du (a), Qi Ye (a), Hong Zhang (a), *, Yu-Guang Zhang (c), **
(a) Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China
(b) Academy of Integrative Medicine, Fujian College of Traditional Chinese Medicine, Fujian 350108, PR China
(c) Department of Orthopaedies, Shanghai Ninth People's Hospital Affiliated lo Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
* Corresponding author. Tel./fax: +86212507454.
** Correspoonding author. Tel./fax: +862163138341.
E-mail addresses firstname.lastname@example.org (H. Zhang), email@example.com (Y.-G. Zhzang).
(1) Contributed equally to this work.
0944-7113/$ -see front matter [c] 2009 Elsevier GmbH. All right reserved. doi: 10.1016.phymed.2009.04.005
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|Author:||Wu, Jian-Guo; Wu, Jin-Zhong; Sun, Lian-Na; Han, Ting; Du, Jian; Ye, Qi; Zhang, Hong; Zhang, Yu-Guang|
|Publication:||Phytomedicine: International Journal of Phytotherapy & Phytopharmacology|
|Date:||Nov 1, 2009|
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