电针对压力性尿失禁大鼠阴道前壁胶原分解与合成的影响

Effect of electroacupuncture on collagen decomposition and synthesis of anterior vaginal wall in rats with stress urinary incontinence

目的观察电针对压力性尿失禁(SUI)大鼠阴道前壁胶原分解与合成的影响。方法60只SD大鼠随机分为假手术组10只、造模组50只。假手术组予开腹处理,找到卵巢组织后直接缝合,不予卵巢切除及阴道扩张;造模组采用阴道扩张后双侧卵巢切除法建立SUI大鼠模型,采用喷嚏试验检验模型是否成功。将造模成功大鼠随机分为模型组、电针非穴位组(简称“针非组”)、电针组、阻断剂组。模型组不予任何处理;电针组大鼠分别在双侧“肾俞”穴和“会阳”穴给予电针治疗,每次20 min,连续3 d;针非组针刺“肾俞”对照点、“会阳”对照点(两穴旁开1 cm)并予电针治疗,每次20 min,连续3 d;阻断剂组予盐酸多西环素灌胃给药,连续3 d。治疗结束后,评价尿动力学指标,取各组大鼠尿道下阴道前壁组织,采用PCR、Western blot、免疫组化的方法检测阴道前壁胶原总含量(Col-Ⅰ、Col-Ⅲ)、降解环节相关因子(MMP-1、MMP-2、MMP-9、TIMP-1)、胶原合成环节相关因子(DCN、TGF-β)的mRNA及蛋白表达。结果(1)造模后,喷嚏试验阳性提示SUI模型造模成功,与假手术组比较,造模大鼠的最大膀胱容量和腹压漏尿点压明显降低(P<0.05)。(2)治疗后,与假手术组比较,模型组大鼠Col-I、Col-ⅢmRNA及蛋白表达水平升高,MMP-1、MMP-2、MMP-9 mRNA及蛋白表达水平降低,TIMP-1 mRNA及蛋白表达水平升高,DCN mRNA及蛋白表达水平降低,TGF-βmRNA表达水平升高(P<0.05)。(3)与模型组比较,电针组及阻断剂组大鼠漏尿点压及膀胱最大容量均升高(P<0.05);电针组阴道前壁Col-I、Col-ⅢmRNA及蛋白表达水平升高(P<0.05),MMP-1、MMP-2 mRNA及蛋白表达水平降低(P<0.05),MMP-9 mRNA表达水平降低(P<0.05)。TIMP-1 mRNA表达水平升高(P<0.05),DCN mRNA及蛋白表达水平降低(P<0.05),TGF-βmRNA表达水平升高(P<0.05)。与针非组比较,电针组Col-ⅢmRNA及蛋白表达水平升高(P<0.05),MMP-1、MMP-2 mRNA及蛋白表达水平降低(P<0.05),MMP-9 mRNA表达水平降低(P<0.05)。TIMP-1 mRNA表达水平升高(P<0.05),DCN mRNA及蛋白表达水平均降低(P<0.05)。结论SUI大鼠阴道前壁胶原蛋白流失,胶原降解增加,胶原合成减少。电针可通过抑制胶原蛋白异常降解,促进胶原蛋白合成,提高SUI大鼠阴道前壁胶原蛋白的总含量,进而达到治疗SUI的目的,且电针穴位疗效优于非穴位。

ObjectiveTo observe the effect of electroacupuncture on the decomposition and synthesis of collagen in anterior vagina wall of rats with stress urinary incontinence(SUI).MethodsTotally 60 SD rats were randomly divided into sham operation group(n=10)and model group(n=50). The sham operation group was treated by laparotomy,and the ovarian tissue was found and sutured directly without ovariectomy and vaginal dilatation;the model group was established by bilateral ovariectomy after vaginal dilatation,and the sneeze test was used to test the success of the model. The successful model rats were randomly divided into the model group, the electroacupuncture non-acupoint group (“acupuncture non-acupoint group”), the electroacupuncture group and the blocker group,and the model group did not receive any treatment. The rats in the electroacupuncture group were treated with electroacupuncture at bilateral“Shenshu(BL23)”and“Huiyang(BL35)”points for 20 min each time for 3 days. The rats in the acupuncture non-acupoint group were treated with acupuncture at the control points of“Shenshu(BL23)”and“Huiyang(BL35)”(1 cm beside the two acupoints) and electroacupunctured for 20 min each time for 3 days,while those in the blocker group were given doxycycline hydrochloride intragastrically for 3 days. After treatment,urodynamic indexes were evaluated,and the mRNA and protein expression of total collagen content(Col-Ⅰ,Col-Ⅲ),degradation link related factors(MMP-1,MMP-2,MMP-9,TIMP-1),collagen synthesis link related factors(DCN,TGF-β)in anterior vagina wall were detected by PCR,Western blot and immunohistochemistry.Results(1) After modeling,the positive sneezing test indicated that the SUI model was successful. Compared with the sham operation group,the maximum bladder capacity and abdominal pressure leakage point pressure of the model rats were significantly lower(P<0.05).(2) After treatment,compared with the sham operation group,the expression levels of Col-I,Col-Ⅲ mRNAs and proteins,TGF-β mRNA,and TIMP-1 mRNA and protein increased(P<0.05),while the expression levels of MMP-1,MMP-2,MMP-9 mRNAs and proteins,and DCN mRNA and protein decreased(P<0.05)in the model group.(3) Compared with the model group,the leak point pressure and the maximum capacity of bladder in the electroacupuncture group and the blocker group were significantly higher than those in the model group(P<0.05). In the electroacupuncture group,the expressions of Col-I,Col-Ⅲ mRNAs and proteins,TIMP-1 mRNA and TGF-β mRNA increased(P<0.05),while the expressions of MMP-1,MMP-2 mRNAs and proteins,MMP-9 mRNA,DCN mRNA and protein decreased(P<0.05)in the anterior vaginal wall. Compared with the acupuncture non-acupoint group,the expression of Col-Ⅲ mRNA and protein and TIMP-1 mRNA increased(P<0.05),while the expressions of MMP-1,MMP-2 mRNAs and proteins,MMP-9 mRNA,and DCN mRNA and protein decreased(P<0.05)in electroacupuncture group.Conclusion collagen synthesis is decreased in the anterior vaginal wall. Electroacupuncture can inhibit the abnormal degradation of collagen,promote collagen synthesis,increase the total content of collagen in the anterior vaginal wall of SUI rats,and then achieve the purpose of treating SUI,and the curative effect of electroacupuncture at acupoints is better than that of electroacupuncture at non-acupoints.