【摘要】：目的探討穴位注射脂肪來(lái)源基質(zhì)血管成分(SVF)聯(lián)合針刺足三里穴位抗皮膚老化作用及其作用機(jī)制。方法采用I型膠原酶溶液及紅細(xì)胞裂解液制備SVF細(xì)胞懸液,染色后采用倒置顯微鏡測(cè)定SVF存活率,采用血球計(jì)數(shù)板細(xì)胞計(jì)數(shù)法進(jìn)行SVF計(jì)數(shù)。24只18月齡SD大鼠隨機(jī)分為四組A組、B組、C組和D組。A組大鼠足三里穴位單次注射PBS, B組大鼠針刺足三里穴位10日,C組大鼠足三里穴位單次注射SVF細(xì)胞懸液,D組大鼠足三里穴位注射SVF細(xì)胞懸液后針刺10日。各組大鼠治療10日脫椎處死后取得各組皮膚組織。各組大鼠脫臼處死后分別取背部部分皮膚組織,勻漿后分別采用黃嘌呤氧化酶法、二甲氨基苯甲醛法、硫代巴比妥酸法和ELISA法測(cè)定皮膚組織SOD活性、羥脯氨酸、丙二醛含量和透明質(zhì)酸水平。各組大鼠取其余皮膚組織常規(guī)石蠟切片,HE染色后比較各組大鼠皮膚厚度；采用兔抗人八因子相關(guān)抗原(Ⅷ-R Ag)多克隆抗體(1：100)行免疫組化染色后比較各組大鼠皮膚血管化程度。結(jié)果(1)B組和C組大鼠皮膚組織SOD活性、羥脯氨酸和透明質(zhì)酸含量均較A組大鼠顯著升高(P＜0.05),B組和C組大鼠皮膚組織丙二醛含量較A組大鼠顯著降低(P＜0.05)；D組大鼠皮膚SOD活性、羥脯氨酸和透明質(zhì)酸含量并且較B組和C組大鼠顯著增加(P0.05),D組大鼠皮膚丙二醛含量較B組和C組大鼠顯著降低(P＜0.05)；(2)B組和C組大鼠皮膚厚度和血管化程度較A組大鼠顯著增加(P0.05)；D組大鼠皮膚厚度和微血管數(shù)目較B組和C組大鼠顯著增加(P＜0.05)。結(jié)論穴位注射脂肪來(lái)源基質(zhì)血管成分(SVF)聯(lián)合針刺足三里通過(guò)增加SOD活性、羥脯氨酸含量,以及降低丙二醛,從而提高皮膚組織的抗氧化防御能力；通過(guò)SVF促進(jìn)皮膚血管化；通過(guò)增加透明質(zhì)酸和膠原蛋白含量,從而提高皮膚厚度；兩者結(jié)合具有更好的抗皮膚老化作用。
[Abstract]:Objective to investigate the anti-aging effect of (SVF) combined with acupuncture at Zusanli acupoint and its mechanism. Methods SVF cell suspension was prepared by using type I collagenase solution and erythrocyte lysis solution. The survival rate of SVF was determined by inverted microscope after staining. 24 18-month-old SD rats were randomly divided into four groups: group A, group B, group C and group D. Group A rats were injected at Zusanli acupoint of group A with single injection of PBS, B for 10 days, and the rats in group B were treated with acupuncture at Zusanli acupoint for 10 days. Group C rats were treated with single injection of SVF cell suspension at Zusanli point and group D rats were treated with SVF cell suspension at point Zusanli for 10 days. The skin tissues of each group were obtained after 10 days of treatment. After the rats were killed dislocated, the skin tissues of each group were removed, and the skin tissue SOD activity and hydroxyproline were measured by xanthine oxidase method, dimethylaminobenzaldehyde method, thiobarbituric acid method and ELISA method respectively. Malondialdehyde content and hyaluronic acid level. The normal paraffin sections of other skin tissues were taken from the rats in each group, and the skin thickness of each group was compared after HE staining. The degree of skin vascularization in each group was compared by immunohistochemical staining with rabbit anti-human factor eight related antigen (鈪，

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