“井穴”放血预处理对急性低氧肺损伤模型大鼠AQP1、AQP5的影响

Effect of Preconditioning of Blood-letting Puncture on Twelve-well of Hand on AQP-1 and AQP-5 of Lung Injury in Rats with Acute Hypoxia

目的观察“手十二井穴”放血预处理对急性低氧肺损伤模型大鼠肺组织水通道蛋白表达的影响,探讨井穴放血预处理对急性低氧肺损伤作用的生物学机制。方法将雄性SD大鼠72只随机分为空白对照组(A)、低氧模型组(D)、“井穴”放血预处理组(C)。模型组及放血组依据低氧时间的不同分为4个亚组,即6h、24h、48h、72h组,每组8只。采用实验动物低氧模拟舱模拟15%氧浓度,制备急性低氧肺损伤模型。放血组大鼠按“少商”“商阳”“中冲”“关冲”“少冲”“少泽”的顺序,采血针放血预处理,每日1次,共5天。采用HE染色法观察肺组织变化,称重计算肺湿干重比,ELISA法检测肺组织TNF-α含量,实时荧光定量PCR(qRT-PCR)技术检测肺组织中AQP1mRNA、AQP5mRNA的表达情况。结果与对照组比较,肺组织形态学观察发现模型组和放血组各时间段大鼠肺组织结构均出现不同程度损伤,与同时间段模型组比较,放血组肺组织损伤程度减轻。与对照组比较,模型组(6h、24h、48h、72h)各时间段大鼠肺湿干重比值均显著增加(P<0.01),肺组织TNF-α含量显著升高(P<0.01),尤以6h升高明显;AQP-1mRNA表达水平增高,在72h时最显著(P<0.01);AQP5mRNA表达则整体呈下降趋势,在24、48、72h时明显(P<0.01)。与同时间段模型组比较,放血组(6h、24h、48h、72h)各时间段大鼠肺湿干重比值较模型组降低(P<0.01);肺组织TNF-α含量降低(P<0.01);AQP1mRNA表达下调,以6h、24h、48h时有统计学意义(P<0.01或P<0.05);AQP5mRNA表达上调,以24h、48h时有统计学意义(P<0.01)。结论“手十二井穴”放血预处理能够在一定时间和程度上改善15%O_(2)急性低氧模型大鼠肺组织损伤,其可能的作用与下调TNF-α含量,调节肺组织内AQP1mRNA、AQP5mRNA的表达,改善肺组织内液体异常积聚,从而减轻炎症反应、减少肺组织的含水量有关。

Objective To observe the effect of bloodletting pretreatment at the"Twelve Well-points"on the expression of aquaporin in the lung tissue of rats with acute hypoxic lung injury,and to explore the biological mechanism of the bloodletting at Well-points on acute hypoxic lung injury.Methods Male SD rats were randomly divided into normal control group A,hypoxia model group D,and"Well-points"bloodletting pretreatment group C which were further divided into 6 h,24 h,48 h,72 h subgroups(n=8 in each group).The experimental animal hypoxia simulation cabin was used to simulate 15%oxygen concentration to prepare the acute hypoxic lung injury model.Group C were pretreated with bloodletting in the order of"Shao Shang"(LU11),"Shang Yang"(LI1),"Zhong Chong"(PC9),"Guan Chong"(TE1),"Shao Chong"(HT9)and"Shao Ze"(SI1),once a day for 5 days.The changes of lung tissue were observed by HE staining method,the wet-to-dry weight ratio of lung was calculated by weighing,the content of TNF-αin lung tissue was detected by ELISA,and the expression of AQP1 mRNA and AQP5 mRNA in lung tissue was detected by qRT-PCR technique.Results Compared with group A,morphological observation of lung tissue showed that the lung tissue structure of rats in groups C and D was damaged to different degrees at each time period.Compared with groups D at the same time period,the degree of C group was reduced.Compared with group A,the wet-to-dry weight ratio of the lungs in group D(6 h,24 h,48 h,72 h)increased significantly(P<0.01),and the content of TNF-αincreased significantly(P<0.01),especially at 6 h;the expression level of AQP-1 mRNA increased significantly,and most significant at 72 h(P<0.01);the expression of AQP-5 mRNA showed a downward trend as a whole,and it was obvious at 24,48,and 72 h(P<0.01).Compared with group D at the same time period,the ratio of wet-to-dry lung weight of rats in group C(6 h,24 h,48 h,72 h)was lower than that in group D(P<0.01);TNF-αcontent in lung tissue was lower(P<0.01);AQP1 mRNA expression was down-regulated,and it was statistically significant at 6 h,24 h,and 48 h(P<0.01 or P<0.05);AQP5 mRNA expression was up-regulated,and it was statistically significant at 24 h,48 h(P<0.01).Conclusion The bloodletting preconditioning of"hand twelve well points"can improve the lung tissue damage of 15%O_(2) acute hypoxic model rats in a certain time and extent.Its possible effect is to down-regulate the content of TNF-αand Regulate the expression of AQP1 mRNA and AQP5 mRNA in lung tissue.Improve abnormal accumulation of fluid in lung tissue,Thereby Reduce inflammation and reduce the water content of lung tissue.