Cannabidiol Activated the Intrinsic Apoptotic Pathway in Murine Lymphocytes

大麻二酚活化小鼠淋巴球之內在凋亡路徑

吳欣穎(Hsin-Ying Wu) ； 廖美秀(Mei-Hsiu Liao) ； 蘇淑婷(Shu-Ting Su) ； 詹東榮(Tong-Rong Jan)

《臺灣獸醫學雜誌》 36卷4期 (2010/12) Pp. 268-278

https://doi.org/10.7009/TVJ.201012.0268

摘要

大麻二酚(cannabidiol; CBD)是大麻成分中主要不具中樞活性的大麻生物鹼，已被報告經由活化外在及內在凋亡路徑誘發神經膠質瘤及血癌細胞之凋亡。我們先前指出CBD在小鼠初代淋巴球細胞所誘發的凋亡和氧化性壓力引起的半胱氨酸蛋白酶－8(caspase-8)之活化有關，顯示CBD會活化外在凋亡路徑。本研究進一步探討CBD誘發小鼠脾臟細胞之凋亡是否涉及粒腺體媒介之凋亡路徑。利用流式細胞儀分析粒腺體之膜電位，結果顯示脾臟細胞給予CBD會造成粒腺體膜電位之去極化，此作用具有時間和濃度相關性。給予CBD處理之脾臟細胞，其Bax和Bak mRNA的表現量增加。此外，隨著CBD處理時間的增加，脾臟細胞之半胱氨酸蛋白酶－9明顯地被活化；給予抗氧化劑N-乙醯－(下標 L)-半胱胺酸(N-acetyl-(下標 L)-cysteine)可有效地降低CBD所誘發之半胱氨酸蛋白酶－9的活化。綜合上述，本研究顯示CBD在初代淋巴球細胞會經由引發氧化性壓力之機制活化內在凋亡路徑。

關鍵字

大麻二酚 ； 內在路徑 ； 半胱氨酸蛋白酶－9 ； 粒線體 ； 脾臟細胞

並列摘要

Cannabidiol (CBD), the major non-psychotropic phytocannabinoid contained in marijuana, has been shown to induce apoptosis in glioma and leukemia cells by activating both the extrinsic and intrinsic pathways. We previously reported that CBD-induced apoptosis in primary lymphocytes was associated with oxidative stress-dependent activation of caspase-8, indicating the activation of the extrinsic pathway. The present studies further investigated the involvement of mitochondrion-mediated apoptotic pathway in CBD-induced apoptosis of splenocytes. Flow cytometric analysis revealed that exposure of splenocytes to CBD resulted in markedly depolarization of the mitochondrial membrane potential (△ψ(subscript m)), in a time- and concentration-dependent manner. The mRNA expression of Bax and Bak was enhanced in splenocytes treated with CBD. In addition, CBD induced the activation of caspase-9 in a time-dependent manner, which was significantly attenuated in the presence of N-acetyl-(subscript L)-cysteine, a thiol antioxidant. Taken together, these results suggest that CBD activates the intrinsic apoptotic pathway via an oxidative stress-dependent mechanism in primary lymphocytes.

並列關鍵字

apoptosis ； cannabidiol ； caspase-9 ； intrinsic pathway ； mitochondria ； splenocyte