【作者】 徐涛；

【导师】 李冀；

【作者基本信息】 黑龙江中医药大学 ， 方剂学， 2014， 博士

【摘要】 目的:通过固金消瘤煎对中晚期肺鳞癌临床及实验研究,探讨固金消瘤煎治疗肺癌的作用机制,为临床提供确有疗效的方药。方法:本次研究的临床部分,将符合纳入标准的60位患者按照随机数字表采用随机单盲分组方法分为治疗组(固金消瘤煎组)30例和对照组(平消胶囊组)30例。疗程2个月,观察固金消瘤煎治疗前后的主要相关症状、瘤体病灶、生存质量和CEA水平以及药物不良反应等变化。实验研究部分,观察固金消瘤煎含药血清作用于SK-MES-1细胞后对其形态学、增殖、凋亡和增殖相关基因CyclinD1蛋白以及凋亡相关基因Bcl-2蛋白和Bax蛋白的影响。结果:(1)治疗组瘤体稳定率为80%,对照组为63.33%,治疗组与对照组稳定率比较,P>0.05,差异无统计学意义,,表明在稳定瘤体方面两组差异无显著性。(2)两组间治疗后比较,经t检验,神疲乏力、口干的症状有显著的统计学意义(P<0.01),咳嗽、气短、食欲不振的症状有统计学差异(P<0.05),而血痰、胸痛的症状无统计学差异,提示固金消瘤煎治疗组在改善神疲乏力、口干、咳嗽、气短、食欲不振方面明显优于对照组,而在改善胸痛、血痰方面两组疗效无明显差异。(3)两组间治疗后卡氏评分积分比较,经t检验,有显著的统计学差异(P<0.01),提示固金消瘤煎治疗组在改善患者生存质量方面明显优于对照组。(4)两组体重方面比较,经R×C列表X2检验,有显著的统计学.差异(P<0.01),提示固金消瘤煎治疗组在增加和稳定患者体重方面较对照组有明显的优势。(5)两组间CEA水平治疗后比较,经t检验,差异有统计学意义(P<0.05),表明固金消瘤煎治疗组在降低患者CEA水平方面明显优于对照组。(6)固金消瘤煎各剂量组含药血清作用于SK-MES-1细胞24h和48h时,部分细胞形体缩小,细胞外形变成不规则形状,黏附力略有降低,可见脱落悬浮于培养液中的细胞。少数细胞肿胀变大、细胞膜破裂呈坏死状。72h时,观察细胞的数量明显减少,而且随着含药剂量的增加贴壁细胞间距逐渐增大,许多细胞体积逐渐变小,形体也随之缩小,细胞的不规则形状也越加明显。坏死的细胞随着增多,表现为形体膨胀,并有破裂。(7)观察固金消瘤煎各剂量组含药血清体外作用于SK-MES-1细胞24小时、48小时和72小时抑制率,其结果显示:固金消瘤煎各剂量组与相同时间的空白血清组相比较,有显著的统计学差异(P<0.01),表明固金消瘤煎对SK-MES-1细胞的增殖抑制作用明显,而且在限定的时间范围内表现出的量-效关系明显。固金消瘤煎高剂量组作用72h后达到了其最大抑制率37.95%。(8)观察固金消瘤煎各剂量组含药血清体外作用于SK-MES-1细胞24小时、48小时和72小时凋亡率,结果表明:固金消瘤煎各剂量组与相同时间的空白血清组相比较,有显著的统计学差异(P<0.05或P<0.01),说明固金消瘤煎对SK-MES-1细胞的诱导凋亡作用明显,而且在限定的时间范围内表现出的量-效关系明显。固金消瘤煎高剂量组作用72h后达到了其最大诱导凋亡率35.27%左右。(9)经固金消瘤煎各剂量组含药血清体外作用于SK-MES-1细胞48h之后,用Western-blot方法检测结果显示,随着固金消瘤煎剂量的增加,Cyclin D1蛋白、Bcl-2蛋白表达逐渐下调,Bax蛋白表达逐渐上调,并且表现出明显剂量依赖性。结论:(1)固金消瘤煎在稳定瘤体方面疗效确切,并能有效改善中晚期肺鳞癌患者的生存质量,增加或稳定患者体重。(2)固金消瘤煎能明显改善神疲乏力、口干、咳嗽、气短、食欲不振等临床症状。(3)固.金消瘤煎对SK-MES-1细胞有明显的抑制增殖作用,在一定的时间内表现出明显的量-效关系。(4)固金消瘤煎能有效下调Cyclin D1蛋白在SK-MES-1细胞中的表达,其可能是抑制增殖作用的机制之一。(5)固金消瘤煎对SK-MES-1细胞有明显的诱导凋亡作用,在一定的时间内表现出明显的量-效关系。(6)固金消瘤煎能显著上调SK-MES-1细胞中Bax蛋白的表达和有效降低Bcl-2蛋白的表达,其可能是诱导凋亡作用的机制之一。更多还原

【Abstract】 Objectives:To study the clinical and experimental effect of the Gujin-Xiaoliu decoction to the middle and late period lung carcinoma.To discuss the mechanism of the Gujin-Xiaoliu decoction to lung carcinoma and provide a effective prescription for clinical therapy.Methods:The clinical part of this study absorbed 60 patients,who were allocated by random single-blinding assignment into two groups,one is therapeutic group(Gujin-Xiaoliu decoction group)and the other is control group(Ping-Xiao capsule group).The course of the treatment are two months and we observe the relevant symptoms,size of the tumor,the quality of life,the level of CEA and drug adverse reaction.The experimental part of this study was to observe the effect of medicated serum of Gujin-Xiaoliu decoction to SK-MES-1 cells through cell morphological effect,proliferation,apoptosis,expression of proliferation-assosiated protein Cyclin D1 expression of apoptosis-assosiated gene Bcl-2and Bax.Results:(1)The proportion of stability of tumor in therapeutic group was 80%,and the proportion in control group was 63.33%.The proportion had no statistical difference between the two groups(P>0.05),which showed therapeutic group didnot have superiority to control group at stabilizing the size of tumor.(2)The comparison through t-test between the two groups after treatment showed thar some symptoms(fatigued spirit and lack of strength,thirst,ect)had significant statistical difference(P<0.01),some symptoms(cough,short of breath,anorexia,etc)had statistical difference(P<0.05),and some symptoms(bloody sputum,thoracalgia,etc)had no statistical difference.The results showed Gujin-Xiaoliu decoction could improve some symptoms such as fatigued spirit and lack of strength,thirst,cough,short of breath,anorexia,but had no superiority to improve bloody sputum and thoracalgia.(3)The kps between the two groups had significant statistical difference(P<0.01)through t-test,this result showed that Gujin-Xiaoliu decoction had superiority to control group at improving the quality of life.(4)The comparision of weight between the two groups had significant statistical difference(P<0.01)through RxC listing X2-test,this result showed that Gujin-Xiaoliu decoction had significant superiority to control group at stabilizing and increasing weight.(5)The comparision of the level of CEA between the two groups had statistical difference(P<0.05)through t-test,this result showed that Gujin-Xiaoliu decoction had superiority to control group at decreasing the level of CEA.(6)We observed the effect of medicated serum of Gujin-Xiaoliu decoction to SK-MES-1 cells at 24h and 48h and found the dimension of some cells were smaller,pyknosis,cellular swelling,membra-nolysis and necrosis.We observed at 72h that the number of cells diminished significantly,and the density of cells diminished along with the higher dose of the medicated serum,a great deal of cells pyknosis,contract.become irregular,and swelling cells,rupture and necrosis cells also increased.(7)The herb serum containing Gujin-Xiaoliu decoction of low,middle and high dose showed growth inhibiting visibly to SK-MES-1 cells at 24h,48h and 72h,the growth inhibiting rate had significant statistical difference(P<0.01),so this result showed Gujin-Xiaoliu decoction had significant growth inhibiting effect to SK-MES-1 cells.And there were dose-dependent and time-dependent manner in some time point.The optimal effective time of herb serum containing Gujin-Xiaoliu decoction on cells was 72hours.The inhibition rate was37.95%.(8)The herb serum containing Gujin-Xiaoliu decoction showed apoptosis inducing ability to SK-MES-1 cells.SK-MES-1 cells were treated by Gujin-Xiaoliu decoction with low、median and high dose medicated serum,and compared with that of control group,there was a significant difference(P<0.01),Gujin-Xiaoliu decoction showed a significant apoptosis inducing effect to SK-MES-1 cells,and the effect showed a dose-dependent and time-dependent manner.The optimal effective time of herb serum containing Gujin-Xiaoliu decoction on cell was 72 hours.The apoptosis inducing rate was about 35.27%.(9)From the results of Western-blot,SK-MES-1 cells were treated by Gujin-Xiaoliu decoction with 48h,the expression of Cyclin D1 and Bcl-2 were reduced,the expression of Bax increased and the effect showed a dose-dependent manner.Conclusions:(1)Gujin-Xiaoliu decoction had superiority at stabilizing the size of tumor,improving the quality of life and stabilizing and increasing weight.(2)Gujin-Xiaoliu decoction could improve some symptoms such as fatigued spirit and lack of strength,thirst,cough,short of breath and anorexia.(3)Gujin-Xiaoliu decoction had significant growth inhibiting effect to SK-MES-1 cells.And there were dose-dependent and time-dependent manner in some time point.(4)Gujin-Xiaoliu decoction decreased the expression of Cyclin D1 of SK-MES-1 cells,which maybe one of the mechanism of Gujin-Xiaoliu’s inhibiting effect.(5)Gujin-Xiaoliu decoction showed a significant apoptosis inducing effect to SK-MES-1 cells,and the effect showed a dose-dependent and time-dependent manner.(6)Gujin-Xiaoliu decoction increased the expression of Bax and decreased the expression of Bcl-2 of SK-MES-1 cells,which maybe one of the mechanism of Gujin-Xiaoliu’s apoptosis inducing effect.更多还原