CAJ | 학술논문

目的：探讨针刺穴位与筋膜在治疗肝纤维化中的体外机制。方法：大鼠分为正常对照组、肝纤维化模型12周组、肝纤维化模型20周组、针刺穴位组和针刺筋膜组。采用HE染色和Mallory染色，光镜下观察各组大鼠肝脏石蜡切片标本，制备各组血清并检测血清中的血清甘油三酯（TG）、透明质酸（HA）、层粘连蛋白（LN）、Ⅲ型前胶原（PC-Ⅲ），验证模型是否造模成功，针刺穴位组和针刺筋膜组是否有效，同时制备各组大鼠血清，对大鼠骨髓间充质干细胞进行干预，以肝细胞标志物白蛋白作为检测指标，采用免疫组化、ELISA法和免疫印迹法检测各组血清对大鼠骨髓间充质干细胞分化增殖是否具有影响。结果：HE染色和Mallory染色及血清学指标显示，造模12周后，大鼠肝脏出现了明显的纤维化和肝细胞的脂肪变，模型组血清中TG、HA、LN、PC-Ⅲ浓度明显高于其他各组。造模20周后，大鼠肝脏出现一定程度自愈，但效果明显差于两个治疗组。免疫组化法、ELISA法和免疫印迹法显示正常对照组血清干预的大鼠骨髓间充质干细胞无白蛋白表达，20周模型组、针刺穴位组和针刺筋膜组干预的大鼠骨髓间充质干细胞均有白蛋白表达。结论：正常对照组血清不能诱导骨髓间充质干细胞向肝细胞分化；而20周模型组、针刺穴位组和针刺筋膜组大鼠血清可诱导骨髓间充质干细胞向肝细胞分化；这提示损伤及治疗均可诱导骨髓间充质干细胞向肝细胞分化。
목적：탐토침자혈위여근막재치료간섬유화중적체외궤제。방법：대서분위정상대조조、간섬유화모형12주조、간섬유화모형20주조、침자혈위조화침자근막조。채용HE염색화Mallory염색，광경하관찰각조대서간장석사절편표본，제비각조혈청병검측혈청중적혈청감유삼지（TG）、투명질산（HA）、층점련단백（LN）、Ⅲ형전효원（PC-Ⅲ），험증모형시부조모성공，침자혈위조화침자근막조시부유효，동시제비각조대서혈청，대대서골수간충질간세포진행간예，이간세포표지물백단백작위검측지표，채용면역조화、ELISA법화면역인적법검측각조혈청대대서골수간충질간세포분화증식시부구유영향。결과：HE염색화Mallory염색급혈청학지표현시，조모12주후，대서간장출현료명현적섬유화화간세포적지방변，모형조혈청중TG、HA、LN、PC-Ⅲ농도명현고우기타각조。조모20주후，대서간장출현일정정도자유，단효과명현차우량개치료조。면역조화법、ELISA법화면역인적법현시정상대조조혈청간예적대서골수간충질간세포무백단백표체，20주모형조、침자혈위조화침자근막조간예적대서골수간충질간세포균유백단백표체。결론：정상대조조혈청불능유도골수간충질간세포향간세포분화；이20주모형조、침자혈위조화침자근막조대서혈청가유도골수간충질간세포향간세포분화；저제시손상급치료균가유도골수간충질간세포향간세포분화。
This study was aimed to discuss cellular mechanism of anti-liver fibrosis by acupuncture on acupoint and connective tissues. Rats were divided into five groups, which were the normal group, liver fibrosis model group (12 week), liver fibrosis model group(20 week), acupoint acupuncture group, and connective tissue acupuncture group. The observation was made on morphology of liver in various groups by HE stain and Mallory stain. Concentrations of TG, HA, LN and PC-Ⅲin serum were also detected. Mesenchymal stem cells (MSCs) were intervened by serum from each group to observe MSCs proliferation and differentiation. The results showed that the serum level of model group(12 week) was significantly different from other groups. And the same results were found in the inspection of morphology. In the normal group there were no cell expressing albumin. In other groups, MSCs can express albumin. It was concluded that the serum of normal group cannot make MSCs transform into hepatocyte-like cell. The serum of other groups can make MSCs transform into hepatocyte-like cell. It was concluded that both hepatic fibrosis and acupuncture can induce MSCs differentiation.