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Induce disease with HgCl2 injections 6. Inject 0.1 ml of 1 mg/ml HgCl2 solution per 100 g rat body weight subcutaneously in different places in the back, flank, or abdominal wall of each animal. Inject control rats with distilled water, 0.9% NaCl, or PBS (the vehicle used for the HgCl2).

Take care to inject HgCl2 strictly subcutaneously, and not intradermally, in order to avoid cutaneous ulcerations or necrosis.

The day of this first injection is considered day 0 of the experiment.

7. Once a week (e.g., at days 7, 14, 21, and 28), collect blood from each rat, process to obtain serum, and determine Ig and autoantibody titers (or freeze for later analysis) as in steps 2 and 3.

Serum IgE and autoantibody titers begin to increase at day 7, peak at days 14 to 21, and then decline.

At days 14 to 21, serum is often lactescent due to hyperlipidemia, a consequence of the nephrotic syndrome.

8. Once a week (e.g., at days 9, 16, 23, and 30), collect urine and measure proteinuria (or freeze for later analysis) as described in step 4.

If blood and urine are collected the same day, let the rats recover after urine collection before bleeding them. High proteinuria (>100 mg/24 hr) may be detected from day 14 on.

9. Every day beginning at day 10, examine rats for clinical manifestations and check for weight loss, mucocutaneous lesions, and the presence of arthritis symptoms (see unit 15.4).

Arthritis has been observed in only a small fraction of rats that receive HgCl2. It is noteworthy that ~50% of rats may die, normally beginning at day 14 after the first HgCl2 injection.

10. At day 15 or 30 euthanize rats (unit 1.8). Process kidneys, freeze in liquid nitrogen, and examine the tissue for the presence of Ig deposits (see Support Protocol 3).

At day 15 rats will predominantly display linear Ig deposits along the glomerular capillary walls; at day 30 glomerular subepithelial deposits will predominate.

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Treating Rheumatoid Arthritis With Herbs Spices Roots

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