黑睛排斥汤防治大鼠高危角膜移植免疫排斥反应的机理研究

Mechanism of Heijing Paichi Decoction in preventing immune rejection on high-risk corneal transplantation rats

目的探讨黒睛排斥汤在大鼠高危角膜移植免疫排斥反应中作用的机理。方法角膜缝线法诱导角膜新生血管,建立大鼠高危角膜移植模型,随机分为A组(模型对照),B组(灌服黒睛排斥汤4 g/(kg.d)),C组(灌服黒睛排斥汤4 g/(kg.d)+CsA 10 mg/(kg.d))。行封闭群大鼠SD-Wistar组间同种异体全板层角膜移植,观察植片存活状况;应用流式细胞分析,免疫荧光组织化学等方法对受体局部和全身免疫状态机制进行检测。结果 1.角膜植片存活时间:A组(10.38±1.69)d,B组(13.00±2.03)d,C组(22.50±3.07)d,3组比较差异有统计学意义(P<0.01)。2.外周血淋巴细胞CD4+和CD8+亚群:术后7 d,CD4+,CD8+,CD4+/CD8+各组之间比较均无明显差别(P>0.05);术后14 d,C组CD4+、CD4+/CD8+与A组比较明显降低(P<0.05);术后21 d,B、C组CD4+、CD4+/CD8+与A组比较明显降低(P<0.05),但各组CD8+比较无明显差异(P>0.05)。3.ConA诱导的脾淋巴细胞的增殖:B组、C组脾T淋巴细胞的增殖被明显抑制,与A组比较差异有统计学意义(P<0.01),其中以C组更为明显。结论黒睛排斥汤对高危角膜移植排斥反应模型大鼠术后免疫排斥反应的抑制作用主要是通过抑制T淋巴细胞的增殖来实现的。

OBJECTIVE To study the action mechanism of Heijing Paichi Decoction on the immunological rejection after high-risk corneal transplantation. METHODS Established homogeneity variant SD-Wister rat＇s high-risk corneal transplantation model. The model was established by suture method including corneal neoangio-genesis and the whole shelf corneal transplantation. Randomly divided into group A-C. Group A ： negative control, Group B ： Heijing Paichi Decoction （4g/kg·d）, Group C ： Heijing Paichi Decoction （4g/kg- d） combined with CsA （ 10mg/kg·d）. We observed the survival status of the allograft and the local and systemic immune status in the recip-ients using immunofluorescence histochemistry and flow cytometry. RESULTS The implant＇s survival time, group A was the shortest, （10.38±1.69d） and group C was the longest （22.5±3.07d）. On the 1st week, the expression of CD4＋ molecule and CD8＋ molecule and the ratio of CD4＋/CD8＋ had no obvious difference in every group （P〉0.05） ; The expression of CD4＋ molecule in experimental B and C was down-regulated on the 2nd week, and it had a significant difference compared with the control group （P〈0.05）. And on the 3rd week, the expression of CD4＋ molecule and the ratio of CD4＋/CD8＋ in experimental B and C was down-regulated compared with the control （P〈0.05）. There was no difference between control and experimental group on CD8＋ expression （P〉0.05）. Group B and C had signifi- cant down-regulation compared with A （P〈0.01）, especially C. The MTT was inhibited in group B and C compared with group A. CONCLUSIONS Heijing Paiehi Decoction has certain effect on preventing high-risk corneal transplantation immune rejection in rat model, which was mainly by inhibiting T lymphocyte proliferation to fulfill.