Curcumin against cisplatin nephrotoxicity and its mechanism
Author: Ying Huang Rong-yi Wang Shunrong
Abstract Objective To investigate curcumin (CMN) Anti-
cisplatin-induced renal injury and its mechanism. Methods 48
male Wistar rats were randomly divided into ? (n = 12), ? (n
= 6), ? (n = 6), ? (n = 12), ? (n = 12) groups, each row
1st intraperitoneal injection of: saline (?, ?, ? group),
CMN 30 mg / kg (? group), CMN 30 mg / kg of zinc
protoporphyrin ? (ZnPP ?) 10 mg / kg (? group), 8 h, from
the ?, ?, ? group of six rats each were killed optional testing HO 1 protein expression and vitality. I mouse again the 2nd intraperitoneal injection of: saline (? group), CMN
20 mg / kg of cisplatin 6 mg / kg (? group). Cisplatin 6 mg /
kg (?, ?, ? group), 5 d after the execution, testing serum creatinine (Cr), blood urea nitrogen (BUN), kidney coefficient, kidney tissue superoxide dismutase (SOD) activity and glutathione GSH (GSH) and malondialdehyde (MDA) content. The results of cisplatin can significantly increase the level of oxidative stress in renal tissue and a serious injury in the kidney structure and function of curcumin only a weak direct anti-cis-platinum renal toxicity, but the pretreatment with curcumin increases HO 1 protein expression and
activity can significantly enhance its anti-cis-platinum renal
toxicity; On the contrary, with ZnPP ? inhibiting HO 1
activity of curcumin could be the elimination of anti-cis-
platinum renal toxicity. Conclusion Curcumin mainly through
increases HO 1 protein expression in an effective
antagonist of cisplatin-induced oxidative stress-induced renal
Key words Cisplatin; curcumin; of heme oxygenase- 1;
oxidative stress; renal toxicity
Study on the Protective Effect and Mechanism of Curcumin on Cisplatin induced Nephrotoxicity
Abstract: Objective To investigate the protective effects of curcumin (CMN) on cisplatin induced nephrotoxicity in
rats and the possible mechanisms. Methods48 male wistar rats
were randomly divided into 5 groups: groupI (n = 12), groupII (n = 6), groupIII (n = 6), groupIV (n = 12), groupV (n = 12). Rats received intraperitoneally injection for the first time respectively with saline (groupI, groupII, groupIII), CMN 30 mg / kg (groupIV), CMN 30 mg / kg Zinc protoporphyrin IX (ZnPPIX) 10 mg / kg (groupV) .8 hours later, half rats in group I, groupIV, groupV were killed to analyse renal HO 1
protein express and activity.Other rats received the intraparitoneally injection for the second time respectively
with saline (groupI), Cisplatin 6mg/kg CMN 30 mg / kg (groupIII) and cisplatin 6 mg / kg (groupII, groupIV, groupV) .5 days later, all the animals were killed to measure the blood creatinin (Cr), urea nitrogen (BUN) and analyse the
renal superoxide dismutase (SOD) activity, glulathion (GSH) and malondialdehyde (MDA) content and histopathological lesion.ResultsCisplatin could increase the renal oxidative stress level and damage seriously the renal histology construction and function.CMN only has slight direct
protective effect against the cisplatin-induced
nephrotoxicity.But the renal protective effect would increased significantly by pretreatment with CMN to up regulate renal
HO 1 protein level and considerably attenuated by inhibiting HO 1 protein activity with ZnPP ?.
ConclusionCMN can protect against cisplatin induced
oxidative stress renal injury by inducing HO 1.
Key words: Cisplatin; Curcumin; Heme oxygenase 1;
Oxidative stress; Nephrotoxicity
Cisplatin is currently the clinical, one of the most commonly used anticancer drugs, such as lung cancer, bladder cancer, testicular cancer and ovarian cancer, such as more
than ten kinds of cancer chemotherapy was recommended as the preferred drug, but the apparent renal toxicity of cisplatin to a large extent would restrict its clinical application. In
recent years, studies have reported that curcumin (curcumin,
CMN) can effectively prevent cisplatin-induced acute renal
injury, but its mechanism has not been fully clarified . In view of cisplatin is mainly through the promotion of free radicals causing oxidative stress suffered kidney damage , while heme oxygenase 1 (HO 1) is the cellular response
to oxidative stress injury in a variety of the most important endogenous protection factor, so we speculated that CMN may be expressed through induction of HO 1 as an anti-cis-platinum
renal toxicity, but has yet to see a related research reports at home and abroad. This article intends to make a study of this.
1 Materials and methods
1.1 Drugs and reagents cisplatin Bio-Pharmaceutical Co.,
Ltd. Yunnan old product, batch number: 060,701. Zinc
protoporphyrin ? (ZnPP ?) and CMN were purchased in Sigma
Corporation. Creatinine (Cr), blood urea nitrogen (BUN), superoxide dismutase (SOD) and protein quantitative detection kit purchased from Nanjing Jiancheng Bio-Engineering
Institute. HO 1 Immunohistochemical detection of antibodies and reagents purchased from Wuhan Boster Company.
1.2 Animal grouping and treatment options 48 healthy male Wistar rats weighing 185 ~ 230 g, were randomly divided into five groups: ? group (control group, n = 12), ? group
(cisplatin alone group, n = 6) , ? group (CMN group, n = 6),
? group (CMN preconditioning group, n = 12), ? group (HO
1 suppression group, n = 12). Respectively, the 1st volume of normal saline via intraperitoneal injection, etc. (?, ?, ?
group), CMN 30 mg / kg (? group), CMN 30 mg / kg ZnPP ? 10
mg / kg (? group), 8 h later, from the ?, ?, ? 3 groups of
6 rats were killed either pick to take kidney detected HO 1
protein expression and vitality. The remaining rats each with the 2nd through the abdominal cavity with normal saline (?
group), CMN 30 mg / kg of cisplatin 6 mg / kg (? group),
cisplatin 6 mg / kg (?, ?, ? group). Usual eating animals
such as free photo of water, 5 d, after weighing, were killed, and renal blood testing of indicators.
1.3 Index Detection
1.3.1 HO 1 were detected by SABC was detected in renal
tissue HO 1 protein expression. Renal tissue in 10% neutral formalin fixed, paraffin-embedded, cut, sliced conventional
dewaxing, to water, 3% H2O2 inactivation of endogenous peroxidase, after antigen retrieval, the normal serum is closed, ? dropping anti-(anti-HO 1 antibody titer of
1:200) and anti-? (titer 1:100), DAB color dark at room
temperature, dehydration, transparent, Fengpian, observed under the microscope, see the cytoplasm there brown granules were as HO 1 positive cells, using CD 8 pathological
color image analysis system at 200 times the light microscope, on each slice were randomly determined 10 not repeat the vision of renal tubular epithelial cells HO 1 protein,
optical density value, averaged on behalf of HO 1 protein
HO 1 activity assay reference to Yao Wood  methods, that is, to take kidney organize about 0.5 g, made of 10% tissue homogenate, high-speed centrifugation microsomal, test Ma Liang Lan quantitative protein concentration, and then under the HO 1 degradation of blood-red The principle
elements of bilirubin generated by measuring the formation of bilirubin in the reaction system on behalf of HO 1 the
amount of energy, units of nmol / (mg protein h).
1.3.2 serum Cr, BUN and kidney biochemical parameters
were measured according to kit instructions were used picric acid method, Determination of diacetyl blood Cr, BUN. Said the kidneys weight, per 100 g kidney weight, the weight of the corresponding coefficient of his kidneys. Take about 1 g renal
cortex tissue homogenate is made, according to kit instructions steps measured SOD activity (xanthine oxidase), GSH (dithio-p-nitrophenyl France), and MDA content
(thiobarbituric France ).
1.4 Histopathological examination taken place small piece
of kidney tissue in 10% neutral formalin fixed, regular alcohol, dimethyl benzene dehydration, embedded in paraffin, sliced, HE staining, light microscope observation of morphological change.
1.5 Statistical analysis of data obtained in this experiment are the measurement data in order to (? s) that
was used to compare between the two groups F q test, using statistical software to complete SPSS11.0. Reposted elsewhere in the paper for free download http://
2.1 kidney HO 1 protein expression and dynamic changes in immunohistochemical analysis showed that, HO 1 protein
was mainly expressed in the cytoplasm of renal tubular epithelial cells, where the basis of group ?, on behalf of
the state, showing rat kidney HO 1 protein expression
volume and activity were lower. ?, ? group had no testing,
but pretreatment with the group ?, so the HO 1 protein
expression and activity should be the same as in group ?. ?
group showed a significant increase in pre-CMN renal HO 1
protein expression and activity. ? group showed ZnPP ?
partially inhibited the CMN-induced HO 1 protein expression and significantly inhibited the HO 1 activity. Figure 1 and Table 1.
Table 1 HO 1 protein expression and activity of the comparison (omitted)
Compared with group ?, * P