EVALUATION OF ANTI-PSORIASIS ACTIVITY OF HERBAL FORMULATION(S) USING IMIQUOMID INDUCED (IMQ) PSORIASIS IN EXPERIMENTAL ANIMALS

                                                                 STUDY PROTOCOL

EVALUATION OF ANTI-PSORIASIS ACTIVITY OF HEARBL FORMULATION(S) USING IMIQUOMID-INDUCED (IMQ) PSORIASIS IN EXPERIMENTAL ANIMALS

1.0 Introduction

Psoriasis is a common skin disorder characterized by the focal formation of inflamed, raised plaques that shed scales from excessive growth of epithelial cells and involve, hyperplasia of epidermal keratinocytes, ectasia, infiltration of T lymphocytes, neutrophils, and other types of leukocytes in the affected skin. Vascular endothelial growth factor (VEGF) is a crucial factor that mediates the angiogenesis of blood vessels and is highly expressed in the skin lesions of psoriasis. VEGF induces microvascular alterations in the dermal papillae, which facilitates the development and persistence of psoriatic lesions.

2.0 TEST SYSTEM DETAILS:

Species                 : Mus musculus (Mouse)

Strain                     : Swiss albino or C57bl/6 or Balb/c

Age                        : 6-10 weeks

Body Wight           : 20-25 g

Sex                        : Male or Female

No. of animals       : 8 /Group

Total animals        : 56 + 6 extra = 62

3.0  ALLOCATION OF GROUPS:

Groups	Treatment	Dose*	Animals
G1	Normal Control	Distilled water	8
G2	Disease Control	Normal saline or 0.25% Na-CMC	8
G3	Reference Drug- Dexamethasone	0.2mg/site	8
G4	Hearbal Formulation-1	X mpk	8
G5	Hearbal Formulation-1	XX mpk	8
G6	Hearbal Formulation-2	X mpk	8
G7	Hearbal Formulation-2	XX mpk	8

*Route of drug administration will be decided at the time of experiment

4.0 METHOD:

·       Healthy animals will be selected, randomized based on body weight and divided into 7 different groups consisting of 8+8 animals each.

·       The dorsal skin of the mice will be shaved and a commercially available

5% IMQ cream will be applied.

·       All groups except normal control will be received a daily topical dose of 62.50 mg of the IMQ cream on the shaved area of their backs for 15 consecutive days. On the right ear, 5% IMQ cream was applied at a dose of 20 mg/cm

·       IMIQUIMOD INDUCED PSORIASIS-LIKE DERMATITIS

·       Treatment will be given as follows:

·       Group G1 will be served as vehicle control.

·       Group G2 will be treated as disease control and stimulated with IMQ 5% on shaved back and right ear pinna (left ear pinna was untreated).

·       Animals of group G3 will be treated with IMQ plus dexamethasone at the dose of 0.2mg/site.

·       Group G4 to G7 animals will be treated with hearbal formulation at different dose levels.

·       Mice will be killed by overdose of thiopentone and 1 punch biopsies will be taken from the treated dorsal skin and weighed in order to measure edema.

·       Biopsies will be then fixed in formalin for histological study as described previously.

·       Alternatively, biopsies will be frozen in liquid N2 and homogenized. Cytokines in skin homogenates were measured by ELISA according to the manufacturer’s specifications, and eicosanoids and MPO.

 

5.0 END POINT PARAMETER(S):

·       Ear Thickness

·       Ear Biopsy weight

·       MPO activity in ear tissue

·       Histopathology of skin tissue

·       Histopathological examination

·       Thickness of right ear pinna

·       The ratio of the weight of the spleen to the bodyweight

·       Assay of cytokine production (IL-17A, IL-22, and IL-23) in

serum and tissue samples

 

6.0  REFERENCE(S):

6.1 Madsen M, Hansen PR, Nielsen LB, Cardoso RM, van Eck M, Pedersen

TX, Imiquimod-Induced Psoriasis-Like Skin Lesions Do Not Accelerate Atherosclerosis in Low Density Lipoprotein Receptor–Deficient Mice, The American Journal of Pathology (2018), doi: 10.1016/j.ajpath.2018.02.005.

6.2 Kent Sakai, Kristen M. Sanders, Marina R. Youssef, Kevin M. Yanushefski, Liselotte Jensen, Gil Yosipovitch, and T. Akiyama. Mouse model of imiquimod-induced psoriatic itch. Pain. 2016 November doi:10.1097/j.pain.0000000000000674. 157(11): 2536–2543.

6.3 Mingkwan N Takuathung, Ariyaphong Wongnoppavich, Ampai Panthong, Parirat Khonsung, Natthakarn Chiranthanut , Noppamas Soonthornchareonnon, and Seewaboon Sireeratawong. Antipsoriatic Effects of Wannachawee Recipe on Imiquimod-Induced Psoriasis-Like Dermatitis in BALB/c Mice. Evidence-Based Complementary and Alternative Medicine. 26 December 2017; https://doi.org/10.1155/2018/7931031, Article ID 7931031, 13 pages.

6.4 Lin Sun, Zeyu Liu, Zibei Lin, Dongmei Cun, Henry HY Tong, Ru Yan, Ruibing Wang, Ying Zheng. Comparison of normal versus imiquimod-induced psoriatic skin in mice for penetration of drugs and nanoparticles. International Journal of Nanomedicine 2018:13 5625–5635

6.5 Jason E. Hawkes, Johann E. Gudjonsson, and Nicole L. Ward. The Snowballing Literature on Imiquimod-Induced Skin Inflammation in Mice: A Critical Appraisal. J Invest Dermatol. 2017 March, doi:10.1016/j.jid.2016.10.024 ; 137(3): 546–549.

                                                                 END OF DOCUMENTS

You may like to read theses links:   

1. List of All SOPs and Documents for In-vivo Laboratory

2. List of All SOPs and Documents for Study Protocol

3. List of Pharmacological Study Models with details about disease Inducing agent, Standard or Reference Drug and related parameters.

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