新疆4种阿魏乙醇提取物抗宫颈癌细胞活性的研究

Study on anti-cervical cancer cell activity of 4 Ferula ethanolextracts from Xinjiang

目的研究新疆4种阿魏(荒地阿魏、多伞阿魏、山地阿魏、准噶尔阿魏)乙醇提取物的抗宫颈癌活性。方法制备新疆4种阿魏根乙醇提取物,以不同浓度(9.375、18.750、37.500、75.000、150.000、300.000 mg/L)干预人宫颈癌Hela细胞,通过MTT法检测吸光度(OD值),求得细胞增殖抑制率及IC_(50),应用流式细胞术分别检测4种阿魏乙醇提取物的细胞凋亡率。取KM小鼠110只,随机分为空白对照组、模型对照组、顺铂组、给药高剂量组、给药低剂量组,除空白对照组外,其他各组将小鼠宫颈癌U14细胞注射于小鼠右肢腋下,造模后第2天灌胃,给药高、低剂量组分别灌胃新疆4种阿魏乙醇提取物1.5 g/kg生药、0.75 g/kg生药,空白对照组、模型对照组灌胃给药相同比例的空白乳剂,连续灌胃12 d,各组末次灌胃1 h后摘眼球取血,取出肿瘤、脾、肾、胸腺,分别测定抑瘤率、脾、肾、胸腺指数,使用全自动生化分析仪器分析测定血清中丙氨酸氨基转移酶、天门冬氨酸氨基转氨酶和胱抑素C的水平,HE染色观察肿瘤组织形态。结果荒地阿魏、多伞阿魏、山地阿魏、准噶尔阿魏对人宫颈癌Hela细胞有抑制作用,IC_(50)分别为25.67、32.67、50.03、54.07μg/mL;对人宫颈癌Hela细胞有促进凋亡作用,凋亡率分别为54.82%、48.99%,51.83%、69.75%。与模型对照组比较,荒地阿魏、多伞阿魏、山地阿魏、准噶尔阿魏高、低剂量给药组瘤重均明显降低,其抑瘤率分别为85.49%、69.15%、29.02%、78.09%、82.16%、55.09%、72.76%、88.67%。与顺铂组比较,新疆4种阿魏瘤重无显著性差异(P>0.05)。与空白对照组比较,模型对照组的脾脏、胸腺明显增大,顺铂组脾脏、胸腺质量降低,山地阿魏、准噶尔阿魏胸腺质量降低,差异有统计学意义(P<0.05),其他阿魏对脾脏、胸腺质量无明显影响;新疆4种阿魏对肾脏质量均无明显影响(P>0.05),新疆4种阿魏对谷丙转氨酶、谷草转氨酶、胱抑素C均无明显影响,顺铂组的胱抑素C值明显升高,差异有统计学意义(P<0.05)。HE染色肿瘤组织,阿魏各给药组肿瘤细胞体积减小。结论荒地阿魏、多伞阿魏、山地阿魏、准噶尔阿魏乙醇提取物具有较强的抗宫颈癌活性。

Objective To study the anti-cervical cancer activity of 4 kinds of ferulaacid(Ferula syreitschikowii,Ferulaferulaeoides,Ferulaakitschkensis,Ferula soongarica)in Xinjiang.Methods The ethanol extracts of Xinjiang Aweigen were prepared and treated with different concentrations(9.375,18.750,37.500,75.000,150.000,300.000 mg/L).The absorbance(OD)of Hela cells were detected by tetrazolium salt colorimetric method,and the cell proliferation inhibition rate and IC_(50) were obtained.Flow cytometry was used to detect the apoptosis rate of 4 ferula ethanol extracts.A total of 110 KM mice were randomly divided into blank control group,model control group,cisplatin group,high-dose administration group and low-dose administration group.Except blank control group,U14 cells were injected into the axilla of the right limb of mice in other groups.On the second day after modeling,the high-dose and thelow-dose groups were given 1.5 g/kg raw drug and 0.75g/kg raw drug of 4 kinds of ferula ethanol extracts from Xinjiang,respectively,while the blank control group and the model control group were given the same proportion of blank emulsion by gavage for consecutive 12 days.The eyeballs were collected 1h after the last administration.The tumor,spleen,kidney and thymus were removed,and the tumor inhibition rate,pleen,kidney and thymus index were measured respectively.The levels of alanine aminotransferase,aspartate aminotransferase and cystatin C in serum were analyzed by automatic biochemical analysis instrument,and the tumor tissue morphology was observed by HE staining.Results The IC_(50) values of Ferulasyreitschikowii,Ferulafeurlaeoides,Ferulaakitschkensis and Ferulasoongarica were 25.67μg/mL,32.67μg/mL,50.03μg/mL and 54.07μg/mL,respectively.The apoptosis rate of Hela cells was 54.82%,48.99%,51.83%and 69.75%.Compared with model control group,more than Ferulasyreitschikowii,Ferulafeurlaeoides,Ferulaakitschkensis and Ferulasoongarica high and low dose group of tumors were significantly decreased,and the inhibitory rate were 85.49%,69.15%,29.02%,78.09%,82.16%,55.09%,72.76%and88.67%.Compared with the blank control group,the spleen and thymus of the model control group were significantly increased,the spleen and thymus of the cisplatin group were significantly decreased,and the thymus of Ferulaakitschkensis and Ferulasoongarica were decreased,and the differences were statistically significant(P<0.05).Other ferulahad no significant effect on the spleen and thymus.The 4 kinds of ferula from Xinjiang had no significant effect on kidney mass(P>0.05),and the 4 kinds of ferula from Xinjiang had no significant effect on alanine aminotransferase,aspartate aminotransferase and cystatin C.The cystatin C value in cisplatin group was significantly increased,and the difference was statistically significant(P<0.05).The tumor tissue was stained with HE,and the volume of tumor cells weredecreased in each ferula administration group.Conclusion The ethanol extracts of ferula from Ferulasyreitschikowii,Ferulafeurlaeoides and Ferulaakitschkensis have strong activity against cervical cancer.