Sunday, 21 June 2020

Studies done with complete Freund's adjuvant




Andrographolide is one of the main active ingredients of Andrographis paniculate, which is one of the Chinese herbal medicines. This ingredient is used in domestic medical treatment for respiratory diseases as well as for inflammation. The main purpose of the study is to probe the effects of andrographolide, by using it as an antioxidant on the oxidative stress level, neutrophil growth and penetration in joints and synovial tissue of arthritis rats stimulated by complete Freund's adjuvant.
The model of rheumatoid arthritis of a rat was induced in the footpad through the subcutaneous injection of complete Freund's adjuvant. After 14 days of induction of the model was established. The treatment was started with different doses, such as 25, 50, 100 mg/kg, of andrographolide and 3 mg/kg of positive control methotrexate from 14th day to 35th day of the clinical treatment. The paw swelling, the hot plate test, arthritis score, histology, and biochemical analysis, were measured to find the effects of andrographolide on oxidative stress, infiltration, and neutrophil accumulation.

From the outcomes of the hot plate test, it was concluded that a high dosage of andrographolide improved the anti-injury capability of rats considerably. The results of the histological and Radiological tests showed that the inflammatory cell infiltration, joint osteoporosis, synovial hyperplasia, as well as other phenomena in the andrographolide group were improved considerably.
Andrographolide slows down the myeloperoxidase as well as the neutrophil elastase activity in arthritis rats stimulated by complete Freund's adjuvant. Thus, it was finally concluded that:

1. Andrographolide decreases neutrophil aggregation in joint tenderness by influencing neutrophil chemotactic factors.
2. Andrographolide lessens the levels of arthritis oxidative stress and it restrains inflammation.


This means that Andrographolide functions as a defensive agent for the treatment of complete Freund's adjuvant-induced rheumatoid arthritis by slowing down lipid peroxidation and nitrate or nitrite levels in a dose-reliant manner, reducing the levels of chemokines as well as inflammatory factors, enhancing antioxidant enzyme activity, and preventing neutrophil buildup and penetration.
Adipose tissue RNA isolation is considered an optimized procedure for reaping high RNA. This is for the reason that the Adipose tissue makeup strongly differs between body areas. Reasonable quantities of unbroken RNA are necessary to examine the local distinctions on a molecular level. Therefore, an optimized isolation method was followed to separate Ribonucleic acid from the samples of adipose tissue.

Excised subcutaneous adipose tissue of an individual was acquired from elective operations, and the RNAlater or RNA Stabilization Reagent was analyzed for its effect on RNA reliability. Additionally, three diverse tissue RNA isolation kits were assessed for effectiveness in isolating Ribonucleic acid from tissue samples. The samples showed a considerable loss in recoverable RNA and RNA degradation signs after 30 to 60 minutes of excising the tissue.

 The application of RNA Stabilization Reagent delayed this degradation considerably. By making use of the RNeasy Lipid Tissue Kit caused a drastically higher RNA yield when compared to using the RNeasy Mini Kit. Thus, it is concluded that combining RNeasy Lipid Tissue Kit and the RNAlater will cause a higher RNA yield even from comparatively small tissue samples with assured RNA integrity.

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