目的:观察逆针灸"关元"穴对自然更年期大鼠免疫功能的影响.方法:经阴道筛查出动情周期紊乱的10月龄SD雌性大鼠160只,随机分为自然10、12、14、16月龄组,逆灸12、14、16月龄组和逆针12、14、16月龄组.另选3.5月龄健康SD雌性大鼠16只为年轻对照组.逆灸、逆针各组从大鼠10月龄开始,每周灸或针"关元"穴2次,共8周.采用免疫组化法检测脾脏热休克蛋白70(HSP 70),原位杂交法检测HSP 70mRNA表达,放射免疫法测定血清白介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)含量.结果:更年期大鼠随着月龄的增长脾脏HSP 70及HSP 70 mRNA的表达呈先升后降的变化,血清IL-2含量波动性降低,TNF-α含量先降后升.与自然同月龄组相比,逆灸、逆针各月龄组HSP 70及HSP 70 mRNA表达和IL-2含量均呈增加趋势,其中逆灸、逆针12月龄组HSP 70及HSP 70 mRNA,逆灸14、16月龄组HSP 70,逆针14月龄组HSP 70 mRNA的表达差异具有统计学意义(P＜0.01或P＜0.05),IL-2含量逆灸14月龄组明显升高(P＜0.05);TNF-α含量逆灸、逆针12月龄组均明显升高(P＜0.01),逆灸16月龄组明显降低(P＜0.05).同月龄逆灸、逆针组相比,逆针12月龄组HSP 70 mRNA显著升高(P＜0.05).结论:逆针灸"关元"穴对调整更年期免疫功能紊乱具有一定作用,其机制可能与调节脾脏组织细胞HSP 70及其基因的表达,启动了机体内在的抗病与应变能力,从而调节血清免疫因子IL-2、TNF-α的含量有关.%Objective To observe the influence of preventive acupuncture (PA) and preventive moxibustion (PM) at "Guanyuan" (CV 4) on the immune function in natural climacteric rats. Methods A total of 160 female SD rats were randomized into control, PA and PM groups, the former one group was further divided into 10 month (mon) ,12 mon, 14 mon and 16 mon subgroups, and the later two groups were further divided into 12 mon, 14 mon and 16 mon subgroups, with 16 rats in each. In addition, other 16 female SD rats aged 3. 5 mon were used as the young control (YC) group. "Guanyuan" (CV 4) was punctured with an acupuncture needle and the needle was retained for 20 min, or given with one ignited moxa-cone from the age of 10 mon on. The treatment was conducted twice every week, 8 weeks altogether. The expression of HSP 70 and HSP 70 mRNA of the spleen tissue was detected by using immunohistochemistry and in situ hybridization respectively, and serum IL-2 and TNF-α contents were assayed by using radio-immunoassay. Results In comparison with YC group, 1) the expression of spleen HSP 70 and HSP 70 mRNA increased significantly in 10 mon control (mon-C), 12 mon-PM and 12 mon-PA groups, and 14 mon-PA group (only HSP 70 mRNA) (P<0. 05, P<0.01); 2) HSP 70 expression decreased remarkably in 14 mon-C, 16 mon-C and 16 mon-PA groups (P<0. 05, P<0. 01); 3) IL-2 contents decreased evidently in 12 mon-C and 14 mon-C groups, and TNF-α contents increased obviously in 12 mon-PM, 12 mon-PA and 16 mon-C groups (P<0. 05). In comparison with the corresponding same age control groups, HSP 70 and HSP 70 mRNA expression increased significantly in 12 mon-PM and 12 mon-PA groups, 14 mon-PM and 16 mon-PM (HSP 70 only), 14 mon-PA (HSP 70 mRNA only) groups (P<0. 05, P<0. 01); IL-2 level of 12 mon-PM group, and TNF-α contents of 12 mon-PM and 12 mon-PA groups increased markedly (P<0. 05, P<0. 01). No significant differences were found between PM and PA groups in most different age groups (P>0. 05). Conclusion Both preventive acupuncture and preventive moxibustion can upregulate the expression of spleen HSP 70 and HSP 70 mRNA and serum IL-2 and TNF-α levels, which may contribute to their effects in enhancing the immune function in menopausal rats.
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