[關鍵詞]
[摘要]
目的 比較瓜蔞薤白半夏湯(GXB)不同提取部位對缺血缺氧心肌細胞的保護作用,初步探尋該經典方劑干預胸痹、心痛的物質基礎。方法 稱取瓜蔞240 g、薤白90 g、半夏120 g,50%乙醇回流提取,分別用石油醚、氯仿、醋酸乙酯、水飽和正丁醇、水5種不同極性溶劑萃取,得到相應提取部位,及藥渣水煎液部位。培養(yǎng)大鼠原代心肌細胞,制備缺血缺氧模型,給予系列濃度(100、10、1、0.1、0.01 mg/mL)的GXB 6種提取部位及瓜蔞皮注射液(陽性對照,100、50、25、5、1 mg/mL)。MTT法檢測細胞存活率,倒置顯微鏡觀察細胞形態(tài)變化,生化試劑盒檢測培養(yǎng)液中肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)活性。結果 與對照組比較,模型組細胞凋亡、壞死明顯,細胞存活率顯著降低,細胞培養(yǎng)液中CK、CK-MB的活性顯著升高(P < 0.05);質量濃度均為0.1 mg/mL的GXB水飽和正丁醇部位、水溶性部位及藥渣水煎部位可以顯著逆轉缺血缺氧心肌細胞的生長狀態(tài)、存活率以及培養(yǎng)液中CK、CK-MB活性(P < 0.05);石油醚、氯仿及醋酸乙酯部位處理的細胞,與模型組無明顯差異。結論 GXB的水飽和正丁醇、水溶性及藥渣水煎部位對缺血缺氧損傷的心肌細胞具有顯著的保護作用。
[Key word]
[Abstract]
objective To compare the protective effect of each part of Gualou-xiebai-banxia Decoction on cardiomyocytes induced by ischemia-hypoxia, and initially explore the material basis of this classical prescriptions intervention on thoracic obstruction and heartache. Methods Weighting 240 g trichosanthes kirilowii, 90 g allium macrostemon and 120 g pinellia ternata for preparation. After 50% ethanol reflux extraction, five different polar solvents including petroleum ether, chloroform, ethyl acetate, n-butanol, and water were used for extraction to obtain their extracts from different sites and decocting part of drug residue. Primary rat myocardial cells were cultured to establish ischemic anoxia model, and a series of concentrations (100, 10, 1, 0.1 and 0.01 mg/mL) of six GXB extracts and gualoupi injection (positive control, 100, 50, 25, 5, 1 mg/mL) were added for treatment. The survival rate of cells was detected by MTT and the morphological changes were observed under inverted microscope, biochemical reagent kit was used to test creatine kinase (CK) and reatine kinase isoenzyme (CK-MB) activity in the liquid culture. Results Compared with control group, the cells in the model group showed apoptosis and necrosis, cell viability significantly decreased, and the activity of CK and CK-MB in cell culture medium increased significantly (P < 0.05). The treatment of water saturated n-butanol site, water soluble site and decocting site of drug residue of 0.1 mg/mL can significantly reverse the growth state, survival rate and myocardial enzyme activity of ischemic and anoxic cardiomyocytes (P < 0.05). However, the cells treated with petroleum ether, chloroform and ethyl acetate had no significant difference with the hypoxia ischemia group. Conclusion The water saturated n-butanol site, water soluble site and decocting site of drug residue of GXB have significant protective effects on myocardial cells injured by ischemia and hypoxia.
[中圖分類號]
[基金項目]
國家自然科學基金項目(81173164、81573852);山東省教育廳高等學校科研項目(J10LF54)