樟芝效用的研究 ：膀胱出口阻塞之缺血傷害病因論

膀胱出口阻塞之缺血傷害病因論 ：樟芝的效用
Robert M. Levin PhD and 林鼎淯 醫師

序言：
膀胱是個平滑肌器官，它的功能是在平時儲存尿液，和經過高度協調的持續收縮來排出尿液；膀胱的功能表現取決於數個因子，包括神經支配、組織、收縮反應、鈣離子的儲存及釋放和新陳代謝的能量。
臨床上，五十歲以上的男人，百分之八十有良性前列腺肥大引起的膀胱排放出口阻塞，最近的證據說明，局部缺血/再灌注是良性前列腺肥大引起的膀胱排放出口阻塞的主要致病因子。我們這個計畫特定目的是探討給予口服樟芝，能保護膀胱經誘導產生局部缺血/再灌注之兩種嚴重傷害兔子的膀胱。

方法：將24隻公紐西蘭大白兔分成4組每組6隻，組別1、2每天餵食含100mg樟芝的懸浮液，組別3、4為標準組，在三星期中所有的兔子均依其組別的每日標準飲食，三星期後，第1及3組的兔子麻醉後手術使其膀胱都受到平均對稱的缺血及一個星期的再灌注，第2及4組的兔子則是對照手術之後給予一星期恢復。在恢復的時間持續按照先前分組之個別標準飲食，在結束後立即進行麻醉和膀胱內壓測量法。膀胱內壓測量法後緊接著立即割去膀胱並秤重。
個別肌肉束藥理學﹕將三個分離取得的膀胱分別置於分離槽，並以氨甲醯醇及氯化鉀來刺激其生理收縮反應。組織學﹕將兩個全皮層束的膀胱體置於10%福馬林溶液中8小時後,作為免疫組織化學研究。另外，我們將神經絲蛋白染色，做為對神經支配效應的標記；以細胞凋零作為細胞損壞的標記。
生物化學方面：將每個膀胱均衡的將膀胱肌肉和膀胱黏膜間隔分離。將其冷凍儲存於-80oC作為生物化學研究。我們測量檸檬酸合成酶活性做為是線粒體功能標記，以肌漿內質網Ca2+ -ATP酶活性作為是肌漿內質網功能標記，和膽鹼乙醯酶活性像是膽鹼能的神經功能標記。


結果：
(1)在控制組，樟芝能增強其膀胱的生理收縮功能，是值得注目的有趣現象。此外，在遭到膀胱缺血/再灌注傷害組，樟芝亦能保護其生理收縮功能。
(2) 在膀胱儲容性上，不論在控制組或缺血/再灌注傷害組，樟芝都具有保護作用。
(3)在膀胱平滑肌，樟芝能在膀胱缺血/再灌注傷害之後，提升肌漿內質網Ca2+ -ATP酶活性（代表粒腺體功能）。
(4) 膀胱缺血/再灌注傷害有顯著減少膀胱平肌的神經密度的破壞現象,而樟芝能在膀胱缺血/再灌注傷害來臨之後，顯著增加神經密度，使之回復到接近正常。
(5) 膀胱缺血/再灌注傷害會顯著引起細胞凋零，而樟芝能顯著減少細胞凋零。

結論：餵食樟芝的兔子有當程度的提升正常膀胱功能的功能表現﹔且在缺血/再灌注的傷害產生時，樟芝能有保護效用。

Ischemic Etiology of Obstructive Bladder Dysfunction:
Effect of Antrodia Camphorata

Robert M. Levin PhD and Alpha DY Lin, MD
Introduction: The urinary bladder is a smooth muscle (SM) organ whose function is to collect and store urine at low intravesical pressure and then to periodically expel the urine via a highly coordinated, sustained contraction. Bladder function depends upon several factors including innervation, structure, contractile response, calcium storage and release, and metabolism of energy. More than 80% of males over 50 years of age have varying degrees of bladder outlet obstruction secondary to benign prostatic hyperplasia (BPH). Recent evidence has demonstrated that ischemia / reperfusion are major etiological factors in bladder dysfunctions secondary to obstructive bladder disease. The specific aim of our project was to determine if orally administered Antrodia camphorata could protect rabbit bladders from the progressive dysfunctions induced by experimental bilateral ischemia / reperfusion.
Methods: 24 male NZW rabbits were separated into 4 groups of 6 animals each. Rabbits in groups 1 - 2 were fed 100 mg Antrodia camphorata suspensions daily; those in groups 3-4 received vehicle. All rabbits received their respective treatments daily for 3 weeks. After 3 weeks, all rabbits were anesthetized and each rabbit in groups 1 and 3 were subjected to bilateral ischemia and one week reperfusion. The rabbits in groups 2 and 4 were sham operated and allowed to recover for 1 week. Feeding was continued during the recovery period. Upon completion each rabbit was anesthetized and cystometry performed. Immediately after cystometry the bladder was excised rapidly and weighed. Isolated muscle strip pharmacology: Three isolated strips from each bladder were mounted in separate baths and the contractile responses to field stimulation (FS), carbachol, and KCl were determined. Histology: Two full thickness strips of bladder body were fixed for 8 hours in 10% buffered formalin and used for immunohistochemical studies. We stained for neurofilament as a marker for innervation and for apoptosis as a marker for cellular damage.
Biochemistry: The balance of each bladder was separated into its muscle and mucosal compartments, frozen and stored at -80oC for biochemical studies. We measured citrate synthase (CS) activity as a marker for mitochondrial function, Sarcoplasmic reticulum Ca2+ ATPase Activiey (SERCA) as a marker for Sarcoplasmic reticulum function and choline acetyltransferase (ChAT) activity as a marker for cholinergic nerve function.
Results: 1) Antrodia resulted in significantly increased contractile responses to all forms of stimulation in both control rabbits and those subjected to I/R. 2) Antrodia resulted in improved bladder compliance in both control and after I/R. 3) In the smooth muscle compartment Antrodia stimulated a marked increase in SERCA activity following I/R. 4) I/R resulted in a significant decrease in the nerve density of the bladder smooth muscle; whereas Antrodia significantly increased the nerve density back toward normal. 5) I/R significantly increased the level of apoptosis; whereas Antrodia reduced apoptosis back to control levels. Conclusion: Feeding rabbits Antrodia camphorata significantly improved normal bladder function; and protected the bladder against significant dysfunctions induced by ischemia / reperfusion.