逆针灸对去卵巢大鼠子宫保护效应的观察

Observation on the Protective Effect of Preventive Acupuncture and Moxibustion on Uterus in Ovariectomized Rats

目的:探讨逆针灸"关元"穴对随后去卵巢更年期模型大鼠子宫保护效应的部分机制。方法:将3.5月龄SD雌性大鼠80只随机分为5组:正常组、假手术组、去卵模型组、逆针组和逆灸组,每组16只。逆针组、逆灸组于大鼠去卵巢之前分别针刺和艾灸"关元"穴,每周2次,共4周,其余3组与逆针、逆灸组同步以手抓放大鼠。至大鼠4.5月龄开始造模,去卵模型组、逆针组、逆灸组行卵巢切除术,假手术组仅切除卵巢周围少量脂肪,正常组不造模。大鼠于造模1个月后(即5.5月龄)取材,采用HE染色、免疫比浊、放射免疫、免疫组化、原位杂交等方法,观察逆针灸对随后去卵巢大鼠子宫组织形态、激素水平、雌激素受体与其基因表达及抗氧化相关指标的影响。结果:大鼠去卵巢后子宫组织形态有明显的衰退表现,子宫内雌二醇(E2)、孕酮(P)含量,雌激素受体(ER)-α及其基因表达,超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性均明显降低,丙二醛(MDA)显著升高(均P<0.01)。与去卵模型组相比,逆灸、逆针组子宫结构的整体情况明显改善,且子宫内P含量、ER-α及基因表达明显升高,MDA显著下降(P<0.05,0.01);同时子宫E2含量,SOD、NOS活性亦有升高趋势,但无统计学意义(P>0.05)。结论:逆针灸"关元"穴对随后去卵巢更年期大鼠子宫退行性变有延缓作用,其保护效应可能与调节子宫局部E2、P含量,ER-α及其基因表达,提高机体抗氧化能力有关。

Objective To observe the effect of preventive acupuncture and moxibustion of ＂Guanyuan＂ （CV 4） on uterus in ovariectomized （OVX） rats. Methods A total of 80 female SD rats were randomly divided into 5 groups： normal control, sham-operation （sham）, model, preventive acupuncture （PA） and preventive moxibustion （PM） groups, with 16 cases in each. PA （with the needle retained for 20 rain after insertion） and PM （one moxa-cone/rat） were applied to ＂Guanyuan＂ （CV 4） sepa- rately before ovariectomy, two times a week, 4 weeks altogether. Then, ovariectomy was performed on rats of model, PA and PM groups. Uterus tissue was taken under anesthesia for homogenate （10 rats/group） or sectioning （5-6 pro, 6 rats/group）. U- terus estradiol （E2）, progestone （P） were detected with radioimmunoassay; superoxide dismutase （SOD）, nitric oxide synthase （NOS）, and malondialdehyde （MDA） were assayed with immunoturdidimetry. Partial uterus slices were stained with H ＆ E method for observing morphological changes, or stained with immunohistochemical method or with in situ hybridization method for displaying estrogen receptor （ER）-α and ER-α mRNA expression separately. Results After ovariectomy, the rat＇s uterus presented obvious squamous metaplasia, hyperplasy, and thickening of the endomembrane, decrease in glands and blood vessels and increase in fibrous connective tissue, etc; while the situation was evidently better in PA and PM groups. In comparison with normal control group, E2, P contents and SOD, NOS activities of uterus tissue in model group decreased significantly （P〈0.01）, while MDA in model group increased evidently （P〈0. 01）. Compared with model group, uterus P content in PA and PM groups in- creased obviously （P〈0. 05）, while uterus MDA decreased apparently （P〈0.01）. Compared with normal control group, the expression of ER-α and ER-α mRNA in model group was downregulated markedly （P〈0.01）; after PA and PM, both ER-α and ER-α mRNA expression increased obviously （P〈0.01）. No significant differences were found between PA and PM groups in the above mentioned 7 indexes （P〉O. 05）. Conclusion Preventive acupuncture and moxibustion of CV4 can postpone the structural degeneration of uterus in OVX rats, which may be related to their effects in modulating the secretion of uterus E2 and P, upregu- lating the expression of ER-α and ER-α mRNA, and improving the anti-oxidative ability.