Effects of Ganoderma Lucidum on Biochemical Dysfunctions of the Rabbit Urinary Bladder using an In-Vitro Model of Ischemia / Reperfusion

Abstract

ABSTRACT:Background: Oxidative stress involving ischemia followed by reperfusion are major etiological factors in obstructive bladder dysfunction in both men and rabbits. Specific natural products such as Ganoderma lucidum with significant antioxidant activity have proven to be useful in their treatment. In two recent studies we demonstrated that pretreatment with Ganoderma was effective preventing bladder dysfunction using both invivo and in-vitro models of ischemia-reperfusion. The current study is a follow-up study using the in-vitro model of ischemia-reperfusion. Methods: Eight New Zealand White rabbits were divided into 2 groups. One group (Group-1) was fed Ganoderma (100 mg/Kg) daily for 3 weeks while the other group (Group-2) were given saline. At the end of a 3 week period, each rabbit was euthanized and the bladder separated into twelve full thickness strips and mounted in individual baths. After 1 hour in oxygenated Tyrodes with glucose, 4 strips from each group were removed frozen and stored at -80 o C. At this time, the oxygenated Tyrodes + glucose were changed to Tyrodes equilibrated with nitro gen in the absence of glucose (ischemia) for 1 hour. After 1 hour of ischemia the buffer was then changed back to normal oxygenated Tyrode's with glucose and allowed to recover for 2 hours (reperfusion). After this time period, the four remaining strips from each group were frozen and stored. The isolated strips were analyzed for the following biomarker enzymes: citrate synthase (mitochondria), calcium ATPase (intracellular calcium movement through the cell wall), and sarcoendoplasmic reticular ATPase (intracellular calcium uptake into the sarcoplasmic reticulum following stimulation. Results: Ischemia-reperfusion resulted in a significant decrease (~50%) in all three enzyme activities. Pretreating with Ganoderma protected all three enzyme activities which were approximately at control levels. Conclusion: Pretreatment of rabbits for three weeks with Ganoderma prior to subjecting them to invitro ischemia-reperfusion significantly protected the bladder enzymes.