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TMR Modern Herbal Medicine  2018, Vol. 1 Issue (3): 127-135    DOI: 10.12032/TMRmhm2017A20
Orginal Articles     
Antioxidative and antiapoptotic effects of (+)-clausenamide on acetaminophen-induced nephrotoxicity in mice
Yu Hong-Min1,2#, Wang Min3#, Yu Zong-Chao4#, Li Yi-Fang1,2, Huang Chun-Xin3, Han Fang-Xuan3,*(), Liu Fan-Na4,*(), He Rong-Rong1,2,*()
1Anti-stress and Health Research Center and Guangdong Engineering Research Center of Chinese Medicine & Disease Susceptibility, Guangdong, China;
2College of Pharmacy, Jinan University, Guangdong, China
3Department of Pharmacy, Hainan General Hospital, Hainan, China
4Department of Nephrology, the First Affiliated Hospital of Jinan University, Guangdong, China.
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Objective: (+)-Clausenamide ((+)-CLA), the active ingredient of wampee, was isolated from the leaves of Clausena lansium (Lour.) Skeels. This study aimed to evaluate the protective potential of (+)-CLA against acetaminophen (APAP)-induced nephrotoxicity in mice. Methods: Mice were divided into control, APAP, high-dose (+)-CLA, and low-dose (+)-CLA groups. Then, mice were preadministered (+)-CLA (50 and 100 mg/kg) for 5 consecutive days. After the last treatment, the animals received a single intraperitoneal injection of APAP (600 mg/kg). Renal histopathology was evaluated by staining with hematoxylin and eosin. The levels of malondialdehyde (MDA) and glutathione (GSH) and the activities of catalase (CAT) and superoxide dismutase (SOD) were determined using corresponding kits. Western blotting was used to analyze the expression of apoptosis-related proteins in renal tissue. Results: Administration of APAP increased serum creatinine and blood urea nitrogen levels in comparison with the control group. An increase in renal MDA level, depletion of GSH, and reductions in CAT and SOD activities in renal tissue indicated that APAP-induced kidney injury was mediated by oxidative stress. The expressions of Bax and caspase-3, cleavage of caspase-3, and cytoplasm cytochrome c levels were up-regulated in renal tissue, whereas Bcl-2 expression and mitochondrial cytochrome c levels were down-regulated in the APAP group, which revealed that APAP-induced kidney injury significantly increased cell apoptosis in renal tubules. The histopathology of kidney tissue supported these biochemical mechanisms. (+)-CLA can reverse changes in most of the abovementioned parameters and nearly restore the normal structure of the kidney. Conclusion: Oxidative stress and apoptosis are considered to be the mechanisms underlying APAP-induced nephrotoxicity. (+)-CLA could be a promising antidote for APAP-induced acute renal damage owing to its antioxidative and antiapoptotic effects.

Highlights

(+)-Clausenamide ((+)-CLA), an acid amide isolated from the leaves of Clausena lansium (Lour.) Skeels, significantly decreases creatinine and blood urea nitrogen levels and increases the antioxidative abilities. The underlying mechanisms of (+)-CLA were involved in improving the antioxidative and antiapoptotic effects. This study provides a basis to clinical application of (+)-CLA.



Key words(+)-Clausenamide      Acetaminophen      Kidney injury      Oxidative stress      Apoptosis pathway     
Received: 23 April 2018      Published: 25 July 2018
Fund:  This work was partly supported by TCM Modernization Program of Hainan Province (2015ZY01), Natural Science Foundation of China (81573675 & 81622050 & 81673709), Science and Technology Program of Guangzhou (201604046016 & 201604020175) and Research Project of Guangdong Provincial Bureau of Traditional Chinese Medicine (20161070).
Corresponding Authors: Han Fang-Xuan,Liu Fan-Na,He Rong-Rong     E-mail: hfx1223@163.com;13560421216@126.com;rongronghe@jnu.edu.cn
Cite this article:

Yu Hong-Min, Wang Min, Yu Zong-Chao, Li Yi-Fang, Huang Chun-Xin, Han Fang-Xuan, Liu Fan-Na, He Rong-Rong. Antioxidative and antiapoptotic effects of (+)-clausenamide on acetaminophen-induced nephrotoxicity in mice. TMR Modern Herbal Medicine, 2018, 1(3): 127-135. doi: 10.12032/TMRmhm2017A20

URL:

https://www.tmrjournals.com/mhm/EN/10.12032/TMRmhm2017A20     OR     https://www.tmrjournals.com/mhm/EN/Y2018/V1/I3/127

Figure 1 Effects of (+)-CLA on renal function.
The levels of Cr (A) and BUN (B) in kidney tissue. The data were expressed as mean ± S.D (n = 6). Data were thought to a statistical significance with *P < 0.05 vs control group; #P < 0.05 vs APAP group.
Cr: Creatinine. BUN: Blood urea nitrogen. APAP: Acetaminophen 600 mg/kg. LCLA: (+)-Clausenamide 50 mg/kg. HCLA: (+)-Clausenamide 100 mg/kg.
Figure 2 Effects of (+)-CLA on renal histopathology.
In APAP group, flat multifocal tubular epithelial cells were pointed out by red arrow. The bar was 10 μm.
APAP: Acetaminophen 600 mg/kg. LCLA: (+)-Clausenamide 50 mg/kg. HCLA: (+)-Clausenamide 100 mg/kg.
Figure 3 Effects of (+)-CLA on antioxidative status.
The levels of GSH (A), SOD (B), CAT (C) and MDA (D) in kidney tissue were shown. The data were expressed as mean ± S.D (n = 6). Data were thought to a statistical significance with *P < 0.05 vs control group; #P < 0.05 vs APAP group.
GSH: Glutathione. SOD: Superoxide dismutase. CAT: Catalase. MDA: Malonaldehyde. APAP: Acetaminophen 600 mg/kg. LCLA: (+)-Clausenamide 50 mg/kg. HCLA: (+)-Clausenamide 100 mg/kg.
Figure 4 Effects of (+)-CLA on Bcl-2, Bax, Caspase-3, Cleaved Caspase-3 and Cytochrome c protein expression.
(A) Western blotting analysis of apoptosis pathway proteins in kidney tissue. (B) The relative intensities of apoptosis pathway proteins were quantified using Image J densitometry software. The data were expressed as mean ± S.D (n = 3). Data were thought to a statistical significance with **P < 0.01, *P < 0.05 vs control group; ##P < 0.01, #P < 0.05 vs APAP group.
APAP: Acetaminophen 600 mg/kg. LCLA: (+)-Clausenamide 50 mg/kg. HCLA: (+)-Clausenamide 100 mg/kg.
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