| Summary: | 碩士 === 高雄醫學大學 === 藥理學研究所 === 101 === Liuwei dihuang wang (LDW), a traditional Chinese herbal formula, consists of Rehmanniae Radix Praeparata (Shu Di Huang), Cornus Fructus (Shan Zhu Yu), Dioscorea Rhizoma (Shan Yao), Alisma Rhizoma (Ze Xie), Poria Cocos (Fu Ling), and Paeonia Cortex (Mu Dan Pi). LDW is traditionally used for treatment of diabetics, renal disorders, and neurosis. Some components of LDW have been confirmed their neuroprotective effects via anti-oxidative, anti-inflammatory, and anti-apoptotic properties. In this study, we aimed to examine the protective effects of LDW on motor neuronal NSC34 cells against survival motor neuron (Smn) protein deficiency. Doxycycline (1 ug/mL) was used to induce Smn deficiency in stable NSC34 cell line carrying Smn-specific shRNA. Cell viability and cytotoxicity were measured by MTT assay and LDH assay, respectively. The condensed chromatin of apoptotic cells were observed by nuclei staining. Protein expressions were determined by western blot analysis. Cell morphology and neurite length were observed by inverted microscope. Our results indicated that Smn-knockdown activated apoptotic pathway, decreased neurite outgrowth and induced neuronal damage. However, LDW (0.01-10 ug/mL) increased cell viability and Smn expression in inducible Smn-knockdown NSC34 cells. Moreover, LDW attenuated apoptotic death via upregulating Bcl-2, reducing cytosolic cytochrome c and decreasing the cleavage of caspase 3. We further investigated the effects of LDW on meurite growth under Smn deficiency. Results indicated LDW promoted neurite outgrowth via inactivation of RhoA/ROCK/p-LIMK/p-cofilin pathway, a negatively regulatory pathway of neurite development. In conclusion, the present results reveal the novel protective effects of LDW against Smn deficiency in NSC34 motor neuronal cells, suggesting the potential therapeutic use of LDW for neurodegerative diseases.
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