食药用真菌粗提物促血管内皮活性的筛选及对高糖损伤内皮的保护作用

Effects of edible-medicinal fungus crude extracts on vascular endothelial cells with high glucose-induced injury

目的:观察古尼虫草、花脸香菇、冬虫夏草和巴西革耳4种食药用真菌粗提物对高糖诱导的人脐静脉内皮细胞(HUVECs)增殖活性的作用,筛选具有保护内皮细胞活性的有效菌菇并探讨其对HUVECs细胞周期及活性氧簇(ROS)水平的影响。方法:体外培养HUVECs,分为对照组(M199正常培养液)、高糖组(33 mmol/L高糖培养液)、4种食药用真菌(古尼虫草、花脸香菇、冬虫夏草和巴西革耳)粗提物干预组(每种粗提物以终浓度12.5、25、50、100 mg/L+高糖培养液),以四甲基偶氮唑蓝(MTT)比色法检测细胞增殖活性,流式细胞术检测细胞周期及细胞内ROS水平。结果:与对照组相比,高糖组HUVECs的MTT吸光度值显著下降,G_0/G_1期的细胞百分率明显增加,S+G_2/M期的细胞百分率明显下降,且ROS水平亦增加(均P<0.05)。与高糖组相比,花脸香菇和巴西革耳粗提物干预降低了细胞吸光度值(P<0.05),且抑制效应呈剂量依赖性增强,而古尼虫草粗提物对其无影响。冬虫夏草粗提物(12.5～50 mg/L)显著提高了HUVECs的增殖活性(P<0,05),减少了G_0/G_1期细胞百分率,提高了S+G_2/M期的细胞百分率(p<0.05),降低了细胞内ROS水平(均P<0.05),且效应呈剂量依赖性增强。结论:4种食药用真菌粗提物经MTT比色法筛选后,仅冬虫夏草粗提物对高糖诱导的HUVECs有显著保护作用,其可能通过促进内皮细胞增殖、进入细胞周期及降低细胞内活性氧水平途径。

AIM：To investigate the effects of crude extracts of Cordyceps gunnii（CGE）,Lepista lentinus （LLE）,Cordyceps sinensis（CSE） and Lentinus striguellus（LSE） on the proliferation of high glucose-treated human umbilical vein endothelial cells（HUVECs）.METHODS：The cultured HUVECs were divided into normal control group （treated with M199 culture medium alone）,high glucose group（treated with M199 culture medium containing 33 mmol/L glucose） and 4 crude extracts of edible-medicinal fungi（CGE,LLE,CSE and LSE ） intervention groups（treated with the crude extract of edible-medicinal fungus at concentrations of 12.5,25,50,100 mg/L in high glucose M199 culture medium）.The cell proliferation was evaluated by MTT assay.The cell cycle and ROS level were measured by flow cytometry. RESULTS：Compared with normal control group,the MTT absorbance value and the percentage of G0/G1 stage of the HUVECs in high glucose group were significantly decreased（P〈0.05 ）,while the percentage of S ＋ G2/M and ROS level were significantly increased（P〈0.05）.Compared with high glucose group,treatment with the crude extracts of Lepista lentinus and Lentinus striguellus decreased the cell absorbance value（P〈0.05 ）,and the inhibitory effect was enhanced in a dose-dependent manner.However,Cordyceps gunnii had no effect（P〉0.05 ）.The crude extracts of Cordyceps sinensis （12.5～50 mg/L） significantly enhanced the proliferative activity of HUVECs,decreased the percentage of G0/G1 stage of HUVECs,increased the percentage of S ＋ G2/M of HUVECs,and reduced the intercellular ROS level（P〈0.05 ）.CON- CLUSION：Only Cordyceps sinensis crude extract effectively protects the HUVECs in high glucose-induced injury,which might be due to promoting more cells to enter to the cell cycle and down-regulating oxidative stress.