- 期刊
基因晶片分析六味地黃丸和桂附八味丸培養腎細胞株基因之不同表現
Microarray Analysis of Genes Differentially Expressed in HEK 293 Cells Cultulred in Rehmannia Six and Eight Formulae
摘要
由於中醫臨床治療腎病分腎陰虛和腎陽虛，本研究嘗試利用基因晶片分析其治療中藥複方，在培養腎細胞株後抽取mRNA，探討基因有何不同之表現。藉腎陰虛和腎陽虛常用方劑－六味地黃丸和桂附八味丸，和DMEM培養胚胎腎細胞株，抽取培養腎細胞mRNA，用中研院白果能博士之基因晶片，作呈色掃描基因表現圖譜分析。結果在六味地黃丸加上，肉桂、附子兩味熱性中藥後之桂附八味九之圖譜分析，其基因表現上調和下降各有20個基因，其中3個基因有顯著意義。1. Cytochrome oxidase V1b (COX V1b) 2. Mevalonate diphosphodecaeboxylase (MDD)。3 Adenine phosphorlbosyltransferase (APRT)。 結論：3個向調控基因，影響內分泌恆定之生化荷爾蒙變化與粒線體能量代謝，和臨床證型顯著相一致。1. COX V1b向上調控粒線體內膜上呼吸酵素，即細胞色素氧化增口，促進粒線體ATP合成。2. MDD酵素於細胞質Peroxisomes內，促進台成酯醇和贍固醇。3. APRT此酵素缺乏的遺傳疾病，大都將導致間質性慢性腎炎腎衰。由以上基因調控瞭解到桂附八味丸治療之機轉，同時中西醫生理功能術語，在基因的解讀中腎陽命門火、腎主納氣和粒線體；腎陰和腎上腺及陸腺得到串聯。命門與三焦清晰權明了人體內組織細胞水分與能量的產生、輸送和代謝。
並列摘要
This holistic concept of traditional Chinese medicine suggests that good health depends on the harmonious functioning of major physiological systems represented by their complementary natures as yin and yang. Representative respectively of yin and yang are mitochondial regulated energy balance and endocrine/tissue water homeostasis. Kidney disorders are accordingly broadly categorized as yin and yang deficient according to their etiological origths. Using the human cell line HEK 293 as substrate and microarray techniques originally developed by Dr Konan Peck of Academia Sinica in which labeled DNA was used for the coupling and estimation of mRNAs, we examined, at the gene expression level, the effects of Liu Wei Di Huang Wan and Ba Wei Di Huang Wan, herbal formulae prescribed for the treatment of kidney-yin and kidney-yang disorders respectively. The results indicated that incubation for 48 hrs in 0.2 mg/mL of Ba Wei Di Huang Wan provided optimal conditions for cell growth while 2 mg/mL was cytotoxic or cytostatic. Treatment with Ba Wei Di Huang Wan fortified with Cortex Cinnamomi and Radix Acconiti led to the up regulation of 20 genes while 20 others were down regulated obviously. Of those up regulated, 3 were compatible with clinical representations. These were respectively 1. Cytochrome oxidase V1b (COXV1b) which up regulats Cytochrome oxidase to increase ATP synthesis, 2. Mevalonate diphosphodecarboxylase (MDD) which stimulates the syntheses of sterol and cholesterol and 3. Adenine phosphoribosyltransferase (APRT) whose defects may lead to interstit 800 Tit8s. Theresults support the concept that the kidney, conceptually visualized as the important Vit 0 Gate and San Jiao points, is a vit 0 link in the Qi system which integrates yin and yang processes and that the herb formulae, through their actions on genes related to the mitochondrial regulated energy balance and endocrine homeostasis, might correct these disorders by optimizing gene expressions.