Effect of Oleo-gum-resin of Boswellia serrata Roxb. (Kundur) on Cadmium Chloride Induced Renal Toxicity in Albino rats
Mahe Alam1, K. Javed2, MA Jafri3
1Central Council for Research in Unani Medicine, Janakpuri, New Delhi-110058, India.
2Department of Chemistry, Faculty of Science, Hamdard University, New Delhi-110062, India.
3Department of Ilmul Advia (Pharmacology), Faculty of Medicine (Unani), Hamdard University,
New Delhi-110062, India.
*Corresponding Author E-mail: mahealam_234@yahoo.co.in
ABSTRACT:
Kundur (oleo-gum-resin of Boswellia serrata Roxb.) is an ingredient in certain Unani formulations used in kidney disorders. Cadmium is potentially a severe nephrotoxin, because cadmium has a biologic half life of more than 30 years. In the present study the effects of Kundur and its methanol soluble (MS) and methanol insoluble (MINS) fractions were investigated on Cadmium chloride induced nephrotoxicity in Albino rats. The animals of group I and II were administered with 1% CMC (1000mgl/kg, p.o.) and the animals of groups III, IV and V were administered with Kundur (1000mg/kg, p.o.), MS (650mg/kg, p.o.) and MINS (350mg/kg, p.o.) respectively for eight days. Animals in group II, III, IV and V were co-administered Cadmium chloride (3mg/kg/bodyweight) from the fourth day to the end of study (eight day). On the 9th day (after 24h s.c. ingestion) blood samples were withdrawal, blood samples centrifuged and serum was separated to measure the concentration of blood urea nitrogen (BUN) and serum creatinine. Albino rats fed with Kundur and MI fraction showed a significant prevention in the rise of serum markers while MS failed to prevent the rise of these serum makers. These results suggest that Kundur (Boswellia serrata Roxb.) and MINS fraction may have potential to reduce the nephrotoxicity in Albino rats.
KEYWORDS: Kundur, Nephroprotective, Cadmium Chloride and Boswellia Serrata.
INTRODUCTION:
Kundur (Oleo-gum-resin of Boswellia serrata Roxb.) is used for the treatment of various ailments such as dysentery, dyspepsia, lung diseases, haemorrhoids, rheumatism, urinary disorders and corneal ulcer in Unani System of Medicine and Ayurvedic System of Medicine for the last several years1-3. It is also an ingredient of certain compound formulations viz Majoon Kundur, Majoon Murawwah-ul-Arwah, Dawa-ul-Kibrit and Habbe Suzak used in Unani medicine for the treatment of different renal disorders4-5.
Various pharmacologically active chemical constituents were isolated from Kundur (Boswellia serrata Roxb.)6-12.
Kundur is known to exhibit antifungal13, anti-complementary14, Juvenomimetic15, anti-inflammatory16-17 and anti-carcinogenic activities18. In advance investigations also revealed that Kundur to possess immunomodulatory properties19, useful in bronchial asthma20, Polyarthritis21, Colitis22, Crohn's disease23 and against Hepatitis C-virus24.
Cadmium is potentially a severe nephrotoxin. Because cadmium has a biologic half life of more than 30 years, nearly all cadmium ingested is retained. The primary nephrotoxicity associated with exposure to this element involves the renal proximal tubule development of a characteristic low molecular tubular proteinuria, calciurea, and in some cases glucosuria. The proximal tubule is the most common component of the nephron to exhibit toxicity from metals but cells in the three distinct anatomical segments (Segments S1, S2 and S3 or P1, P2 and P3) vary greatly in their response to a given metal. Cadmium is reported to elicit toxicity in only the S1 and S2 segments.25 The S1 segment comprises the initial portion of the proximal convoluted tubule while the end of the convoluted segment and the initial portions of the straight segments in the medullary ray compose S2 segment.
The nephrotoxicity of cadmium (Cd) in man and experimental animal models has been known for many years26-28 and there have been a number of extensive reviews29-31. The primary nephrotoxicity associated with exposure to this element involves the renal proximal tubule development of a characteristic low molecular tubular proteinuria, calciuria, and in some cases glucosuria. The low molecular weight metal binding protein metallothionein appears to play a central role in the mechanisms underlying these phenomena.
Mammalian metallothionein (MT) is a highly conserved protein with a molecular mass of approximately 6,000-10000-d that is composed of approximately 30% cysteine residues. A number of metals such as cadmium, zinc, copper, mercuric ion, and silver may induced and bind MT. The cysteine residues of this protein act to bind seven metal atoms by forming two metal-binding clusters of four and three metals, respectively, with four cysteines per Cd through a bridging sulfur bond.32 Metallothionein synthesized in the liver following exposure to Cd has been shown by numerous researchers to be present in the circulation. As a low molecular weight protein, MT is rapidly and efficiently filtered by the glomerulus and reabsorbed by the renal proximal tubule cells. Where it is rapidly degraded by the lysosome system with release of Cd+, which turn cause the synthesis of renal MT shows a representation of this process and the known and hypothesized relationships between this process and tubule cell injury33-50.
Materials and methods:
Kundur used in the current study, was obtained from Qadimi Unani Dawakhana Ballimaran, Delhi, India. The authenticity of Kundur was established as Oleo-gum-resin of B. serrata by Prof. S. H. Afaq and Dr. Mohinul Haq Siddiqui, Dept. of Ilmul Advia, Ajmal Khan Tibbiya College, Aligarh Muslim University, Aligarh. The voucher specimen MA-K-02-03 of this drug was preserved at the Department of Ilmul Advia, Faculty of Medicine, Hamdard University, New Delhi, India.
Albino Wistar rats of either sex weighing 175-250 gm were used in the present study. The animals were obtained from Jamia Hamdard University, New Delhi and the Animal Ethical Committee of Jamia Hamdard University approved the study protocol. The animals were randomly assigned to five separate groups. Six animals were allotted to each group. Throughout the study all animals were kept under standard laboratory conditions: Temperature 28+1°C, and 12 hr light/ dark cycle. The experiments were performed between 09:00 and 17:00 hours. The animals were fed with standard diet (supplied by the New Maharashtra Chakan oil Mills Ltd., Mumbai) and water ad libitium.
Kundur was dried over calcium chloride (CaCl2) in a desiccator under reduced pressure. The dried oleo-gum-resin was crushed thoroughly and extracted with methanol by refluxing on boiling water bath for 10 -15 minutes. It was filtered and the residue was further extracted with methanol two times. All the filtrates were combined together and the solvent was removed under reduced pressure. The extract obtained after removal of methanol was named as MS (Methanol soluble). The residue (Methanol Insoluble) left on the filter paper was named as MINS. The yields of MS and MINS were found to be 65% and 35% respectively.
Carboxymethyl cellulose (CMC) and methanol were procured from Central Drug House, New Delhi, India, Cadmium Chloride was obtained from Central Drugs House Pvt. Ltd. India. The drugs were suspended in distilled water with 1% Carboxymethyl cellulose (CMC).
Experimental procedure:
The current study was designed for eight days.
Group I: was given vehicle (1% CMC in D.W, 10ml/kg/day, p.o) and served as control. Distilled water was abbreviated as D.W. and body weight as b.w.
Group II: was given vehicle (1% CMC in D.W 10ml/kg/d, p.o.) and Cadmium Chloride (3mg/kg) for 8 days. It was toxic group.
Group III: The animals in group-III received Kundur (1g/kg b.w/d) suspended in vehicle (10ml/kg) and Cadmium Chloride (3mg/kg) for eight days.
Group IV: The animals received methanol soluble (MS) Kundur fraction (650mg/kg/b.w/d) suspended in vehicle (10ml/kg) with Cadmium Chloride (3mg/kg) for eight days.
Group-V: The animals received methanol insoluble (MINS) Kundur fraction (350mg/kg b.w/d) suspended in vehicle (10ml/kg) with Cadmium Chloride (3mg/kg) for eight days.
The animals in group II, III, IV and V were co-administered Cadmium Chloride (3mg/kg) dissolved in normal saline from the 4th day up to the end of the study (eight day). The dose of Cadmium Chloride (3mg/kg) was injected subcutaneously in neck region in a volume of 1ml/kg (10% concentration). Group I received normal saline instead of Cadmium Chloride (3mg/kg) as suggested earlier41. On the 9th day, blood samples from each animal were collected and analyzed for blood urea and serum creatinine levels following the standard procedure51.
All the values were expressed as mean ± S.E.M. Student’s t-test was used to analyze significance of the two means. Probability level of less than 5% was considered as statistically significant.
The effect of Oleo-gum-resin of Boswellia serrata Roxb, (Kundur) and its fractions MS and MINS on renal function was examined in Cadmium Chloride nephrotoxicity model. The daily subcutaneous administration of Cadmium Chloride at 3mg/kg for 8 days caused renal dysfunction in the rats as evidenced by marked elevation in blood urea (431.33%) and serum creatinine (225.24%) compared to saline-treated (s.c) animals. Co-administration of Kundur (1gm/kg, p.o.) with cadmium chloride (s.c.) inhibited the rise of urea (65.44%) and serum creatinine (127.76%). Kundur was fractionated into methanol soluble (MS) and methanol insoluble (MINS) fractions. These fractions (MS & MINS) equivalent to 1g/kg of Kundur were also studied. There were 65.95% inhibition in the rise of BUN and 99.35% inhibition in the rise of serum creatinine with MS while 67.23% and 123.31% respectively with MINS. (Table 1)
Table 1: Effect of Oleo-gum-resin of Boswellia serrata, (Kundur) and its fractions (MS and MI) on BUN and creatinine in Cadmium Chloride induced nephrotoxicity
|
Groups |
Dose Treatment (8 days) |
N |
BUN (mg/dl) Mean ± S.E.M. |
Creatinine (mg/dl) Mean ± S.E.M. |
|
I |
CMC (1000mg /kg) |
6 |
6.16 + 0.38 |
2.06 + 0.08 |
|
II |
CMC+ Cadmium Chloride (3 mg/kg) |
6 |
26.57 + 1.17***a |
4.64 + 0.21***a |
|
III |
Kundur (1 gm/kg)+ Cadmium Chloride (3 mg/kg) |
6 |
12. 54 + 0.45***b |
1.34 + 0.07***b |
|
IV |
MS (650 mg/kg)+ Cadmium Chloride (3 mg/kg) |
6 |
12.44 + 0.39***b |
2.08 + 0.12***b |
|
V |
MINS (350 mg/kg)+ Cadmium Chloride (3 mg/kg) |
6 |
12.21 + 0.32***b |
1.46 + 0.03***b |
***P<0.001 statistically significant (a vs. control and b vs. a).
NS (Statically more significant). Student’s t-test.
N = Number of animals in each group.
CMC (10 ml/kg), Cadmium Chloride (3 mg/kg) Kundur (1000mg/kg), MS (650mg/kg), and MI (350mg/kg) doses were given to the above-mentioned groups.
In the present study, cadmium chloride treatment caused nephrotoxicity as evidenced by marked elevation in blood urea (431.33%) and serum creatinine (225.24%) compared to saline-treated (s.c) animals. Co-administration of Kundur (1gm/kg, p.o.) with cadmium chloride (s.c.) inhibited the rise of urea (65.44%) and serum creatinine (127.76%). Kundur was fractionated into methanol soluble (MS) and methanol insoluble (MINS) fractions. These fractions (MS & MINS) equivalent to 1g/kg of Kundur were also studied. There were 65.95% inhibition in the rise of BUN and 99.35% inhibition in the rise of serum creatinine with MS while 67.23% and 123.31% respectively with MINS. There are many plants and its products which regulates the cadmium chloride induced nephrotoxic effect. The Withania somnifera L (Dunal), Asgand, Rheum emodi Wall ex Meissn. (Revandchini), Tribulus terrestris L (Khar-e-Khasak Khurd) and Dolichos biflores (Bunge) Regel and Ferula foetida Regel Kulthi have been reported to possess protective effect against cadmium induced nephrotoxicity in animal models respectively52-58.
CONCLUSION:
The results of our current study investigation revealed that essential oil of Kundur and the chemical constituents of its methanol soluble fraction are capable to reduce the nephrotoxicity caused by Cadmium Chloride. However, to reach any conclusive decision before recommending Kundur in cases of renal disorders detailed phyto-pharmaco-toxicological studies are necessary to identify the active principle of Kundur and their exact mechanism of action.
CONFLICT OF INTERESTS:
The authors have no conflicts of interest to disclose.
FUNDING/ SUPPORT:
The authors received no funding sources/grants or other materials supports for this works.
ACKNOWLEDGMENT:
None
ABBREVIATIONS:
Cd: Cadmium
CMC: Caboymethyl cellulose
D.W: Distilled Water
MINS: Methanol Insoluble
MS: Methanol soluble
MT: Mammalian metallothionein
P.O.: Per orally
S.C: Subcutaneous
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Received on 04.10.2022 Modified on 08.11.2022
Accepted on 05.12.2022 ©Asian Pharma Press All Right Reserved
Asian J. Pharm. Tech. 2023; 13(3):189-193.
DOI: 10.52711/2231-5713.2023.00034