银杏叶提取物联合胶原蛋白抑制皮肤鳞状细胞癌的动物学机制研究

【作者】 刘楠；

【导师】 白彦萍；

【作者基本信息】 北京中医药大学 ， 中西医结合临床（专业学位）， 2020， 硕士

【摘要】 本文主要包括文献综述及实验研究两部分。文献综述部分:文献综述一:银杏叶提取物药理作用研究进展文献综述二:胶原蛋白应用的研究进展实验研究部分:目的:1.阐明7-12二甲基苯并蒽联合UVB照射是否可以快速诱导建立小鼠皮肤鳞状细胞癌模型。2.阐明银杏叶提取物及胶原蛋白是否可以通过JAK2/STAT3/SOCS1通路抑制皮肤鳞状细胞癌的发生,从靶向、靶标方面,为银杏叶提取物及胶原蛋白的临床应用提供客观的实验依据。方法:1.BALB/c小鼠分组后背部剃除毛发,分别联合应用DMBA/UVB与单用DMBA处理小鼠背部皮肤,每周三次,持续16周。使用数码相机记录小鼠背部皮损变化情况;HE染色检测皮损中鳞状细胞癌发生情况;PCNA免疫组织化学染色检测皮损中恶性程度的高低。2.BALB/c小鼠分组后背部剃除毛发,在应用DMBA/UVB处理小鼠背部皮肤的同时分别使用银杏叶提取物、胶原蛋白与联合应用银杏叶提取物及胶原蛋白进行干预,每周三次,持续16周。使用数码相机记录小鼠背部皮损变化情况;HE染色检测皮损中鳞状细胞癌发生情况;PCNA免疫组织化学染色检测皮损中恶性程度的高低;并使用RT-PCR法检测各组皮损中JAK2、STAT3及SOCS1的表达情况。结果:1.在小鼠皮肤鳞状细胞癌的模型构建实验中,相较于单纯使用DMBA,联合应用DMBA/UVB的小鼠有着肿瘤发生迅速、肿瘤数目多、肿瘤体积大等特点,结合HE染色及PCNA免疫组织化学染色,提示模型构建成功。2.在银杏叶提取物及胶原蛋白抑制小鼠皮肤鳞状细胞癌发生的实验中,模型组联合应用DMBA/UVB处理小鼠背部皮肤后,在红斑的基础上逐渐出现丘疹,且丘疹逐渐增多、增大且相互融合,HE染色提示造模成功。银杏叶提取物、胶原蛋白及银杏叶提取物与胶原蛋白联合组小鼠背部皮损较模型组有着不同程度的好转,HE染色及PCNA免疫组织化学染色均提示银杏组与联合组小鼠皮肤鳞状细胞癌恶性程度低于模型组。PCR法发现实验组小鼠皮肤JAK2与STAT3的表达较对照组升高,但显著低于模型组,SOCS1的表达低于实验组,但高于模型组。结论:1.实验一说明相较于单纯使用DMBA,联合应用DMBA/UVB可以更加快速稳定地构建小鼠皮肤鳞状细胞癌模型。2.实验二说明银杏叶提取物可以通过调控JAK2、STAT3及SOCS1的表达来起到抑制皮肤鳞状细胞癌的作用。单独使用胶原肽可以在一定程度上抑制小鼠皮肤鳞状细胞癌的发生。胶原肽与银杏叶提取物的联合应用并不能加强银杏叶提取物的抑制皮肤鳞状细胞癌发生的作用。更多还原

【Abstract】 This dissertation is composed of two parts:literature review and experimental research.The part of literature review:The first literature review:Advances in pharmacological activities of ginkgo biloba extractThe second literature review two:Advances in the application of collagen proteinOBJECTIVE:1.To clarify whether 7-12 dimethyl benzanthracene combined with UVB irradiation can rapidly induce the establishment of mouse skin squamous cell carcinoma model.2.Clarify whether ginkgo biloba extract and collagen can inhibit the occurrence of skin squamous cell carcinoma through JAK2-STAT3-SOCS1 pathway,and provide objective experimental basis for the clinical application of ginkgo biloba extract and collagen from the aspects of target and target.METHODS:1.BALB/c mice were divided into groups and shaved on the back.DMBA/UVB and DMBA alone were applied to the back skin of mice three times a week for 16 weeks.A digital camera was used to record the changes in the skin lesions on the back of mice.HE staining was used to detect the occurrence of squamous cell carcinoma in the skin lesions,and PCNA immunohistochemical staining was used to detect the degree of malignancy in the skin lesions.2.BALB/c mice were divided into groups and shaved on the back.While DMBA/UVB was applied to treat the skin on the back of the mice,ginkgo biloba extract and collagen were used and combined with ginkgo biloba extract and collagen for intervention three times a week for 16 weeks.A digital camera was used to record changes in back lesions.HE staining was used to detect squamous cell carcinoma in skin lesions.PCNA immunohistochemical staining was used to detect the degree of malignancy in skin lesions.The expression of JAK2,STAT3 and SOCS1 in each group was detected by RT-PCR.Results:1.In the construction experiment of skin squamous cell carcinoma in mice,compared with DMBA alone,the mice with combined DMBA/UVB showed the characteristics of rapid tumor development,large number of tumors and large tumor volume.The combination of HE staining and PCNA immunohistochemical staining indicated the success of the modeling.2.In the experiment of ginkgo biloba extract and collagen inhibiting the occurrence of squamous cell carcinoma in the skin of mice,papules gradually appeared on the basis of erythema after DMBA/UVB treatment in the model group,and papules gradually increased,increased and fused with each other.HE staining suggested the success of modeling.Compared with the model group,the dorsal skin lesions of mice in the combination group of ginkgo biloba extract,collagen and ginkgo biloba extract and collagen had different degrees of improvement.HE staining and PCNA immunohistochemical staining indicated that the malignant degree of skin squamous cell carcinoma in the ginkgo group and the combination group was lower than that in the model group.PCR method found that the expression of skin JAK2 and STAT3 in the experimental group was higher than that in the control group,but significantly lower than that in the model group,and the expression of SOCS1 was lower than that in the experimental group,but higher than that in the model group.Conclusion:1.Experiment 1 shows that compared with DMBA alone,combined application of DMBA/UVB can construct mouse skin squamous cell carcinoma model more quickly and stably.2.Experiment 2 showed that ginkgo biloba extract could inhibit squamous cell carcinoma by regulating the expression of JAK2,STAT3 and SOCS1.Collagen peptide alone can inhibit the development of squamous cell carcinoma in mice to some extent.The combination of collagen peptide and ginkgo biloba extract could not enhance the inhibitory effect of ginkgo biloba extract on squamous cell carcinoma.更多还原