EVALUATION OF ACTIVITY OF HERBAL FORMULATIONS IN RAT MODEL OF STREPTOZOTOCIN INDUCED DIABETIC CATARACT

EVALUATION OF ACTIVITY OF HERBAL FORMULATIONS IN RAT MODEL OF STREPTOZOTOCIN-INDUCED DIABETIC CATARACT

1.0 TEST SYSTEM DETAILS:

Species : Rattus norvegicus (Rats)

Strain: Wistar or Sprague Dawley

Age: 8-9 weeks

Sex: Male

No. of animals: 10 /Group

Total animals: 70 Males

2.0 TEST ARTICLES DETAILS

HF1:  Herbal Formulation -1

3.0 ALLOCATION OF GROUPS:

Group No.	Group Description	Disease Induction procedure	Treatment administered	Dose Volume and Route
G1	Normal Control	0.1 M citrate buffer, pH 4.5	0.5% MC, p.o., b.i.d.	5 ml/kg, p.o.
G2	Disease Control	Single intraperitoneal injection of Streptozotocin (65 mg/kg) in 0.1 M citrate buffer, pH 4.5	0.5% MC, p.o., b.i.d.
G3	Reference Control		Resveratrol-30 mg/kg, p.o., b.i.d.
G4	Treated with a low dose of HF1		HF1: 5-15 mg/kg, p.o., b.i.d.
G5	Treated with intermediate dose 1 of HF1		HF1: 15-50 mg/kg, p.o., b.i.d.
G6	Treated with intermediate dose 2 of HF1		HF1: 50-150 mg/kg, p.o., b.i.d.
G7	Treated with a high dose of HF1		HF1: 150-500 mg/kg,  b.i.d.

4.0 METHOD:

• Healthy animals will be selected for the study, randomized based on body weight, and assigned to 7 groups of 10 animals each.

• Animals of Group G1 will be designated as normal-control and administered 0.5% MC, p.o., b.i.d.

• Disease control animals (assigned to group G2) will receive 0.5% MC, p.o., b.i.d.

• Animals of group G3 will be treated with reference drug Resveratrol 30 mg/kg, twice daily.

• Animals of group G4-G7 will be treated with HF-1, at different dose levels ranging from 5-500 mg/kg, b.i.d.

• The animals allocated to groups G2-G7 will be administered with a single injection of Streptozotocin- 65 mg/kg BW, intraperitoneally for induction of diabetes and subsequently cataract. Animals of G1 will be administered with a single injection of 0.1 M citrate buffer, pH 4.5. Additionally, animals of groups G2-G7 will receive 5% dextrose in drinking water to accelerate the development of cataracts.

• Compound administration will be initiated 2 weeks after streptozotocin administration and continued till the end of the study.

• Eight weeks after disease induction, animals will be sacrificed under an overdose of thiopentone anesthesia. After suitable anesthesia but before the animal dies, blood will be collected from the retro-orbital plexus for the estimation of biochemical parameters. Immediately after the animal dies, the eye lens will be collected and stored at -80°C for evaluation of different parameters.

5.0 PARAMETERS TO BE EVALUATED:

• Body weight: Twice a week.

• Blood Glucose level- once weekly

• Insulin level measurement terminally

• Eye examination and grading for appearance of cataract- once weekly

• SDS-PAGE examination of lens proteins

• Gene expression analysis of eye lens by Real-Time PCR: iNOS, NF-κB, Il-1β, and VEGF

• Oxidative stress analysis in lens tissue- Glutathione peroxidase, superoxide dismutase, Malondialdehyde.

6.0 REFERENCES:

1. Chen, Y., Sun, X. B., Lu, H. e., Wang, F. & Fan, X. H. Effect of luteoin in delaying cataract in STZ-induced diabetic rats. Arch. Pharm. Res. 40, 88–95 (2017).

2. Suryanarayana, P. et al. Digital Commons @ Becker Emblica officinalis and its enriched tannoids delay streptozotocin- induced diabetic cataract in rats Please let us know how this document benefits you . Emblica officinalis and its enriched tannoids delay streptozotocin- induced diabetic cataract in rats. (2007).

3. Raju, T. N., Kanth, V. R. & Lavanya, K. Effect of methanolic extract of Allium sativum (AS) in delaying cataract in STZ-induced diabetic rats. J. Ocul. Biol. Dis. Infor. 1, 46–54 (2008).

4. Valavala, V. K., Vangipurapu, R. K., Banam, V. R., Pulukurthi, U. M. R. & Turlapati, N. R. Effect of mustard (brassica juncea) leaf extract on streptozotocin-induced diabetic cataract in Wistar rats. J. Food Biochem. 35, 109–124 (2011).

                                                           END OF THE DOCUMENT

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