腹腔注射腺病毒pMultiRNAi-Ldhc对高原鼠兔骨骼肌Ldhc基因的沉默效果

The Silence of Ldhc in Plateau Pika Skeletal Muscle by Intraperitoneal Injection of Adenovirus pMultiRNAi-Ldhc

腹腔注射是一种简单且方便给药的方式,为了验证腹腔注射腺病毒pMultiRNAi-Ldhc对高原鼠兔(Ochotonacurzoniac)骨骼肌Ldhc基因沉默的可行性,将27只高原鼠兔分为干扰组、空壳组和空白对照组,每组各9只个体,干扰组和空壳组分别注射0.65ml腺病毒pMultiRNAi-Ldhc和腺病毒pMultiRNAi-NS,空白对照组注射等量生理盐水,注射后7d检测骨骼肌中Ldhc基因mRNA和蛋白的表达水平,测定了乳酸脱氢酶(LDH)活性和乳酸(LD)及三磷酸腺苷(ATP)的含量。结果表明,与空白对照组相比,干扰组在mRNA和蛋白水平上,Ldhc基因表达分别降低了41.73%和15.76%;乳酸脱氢酶(LDH)活性、乳酸(LD)和ATP含量分别降低了23.98%、51.08%和19.29%。结果说明,腹腔注射腺病毒pMultiRNAi-Ldhc能有效沉默骨骼肌中Ldhc基因表达。

Plateau Pika（Ochotona curzoniac） is a small mammal native to Qinghai-Tibet area of 3 000﹣5 000 m above sea level. Previous studies found that the sperm-specific lactate dehydrogenase（LDH-C4） gene is expressed in Plateau Pika skeletal muscle. Intraperitoneal injection is a simple and convenient method for drug injection. In order to verify the effect of Ldhc silence in Plateau Pika skeletal muscle by intraperitoneal injection of adenovirus p Multi RNAi-Ldhc as a method of RNAi, 27 Pikas were randomly divided into the RNAi-LDHC, RNAi-HK and control groups, each had nine individuals. The Pikas of RNAi-LDHC, RNAi-HK were injected with 4 × 109 pfu 0.65 ml adenovirus p Multi RNAi-Ldhc and p Multi RNAi-NS, respectively. The control group was injected the same amount of normal saline. Seven days after injection, swimming time and the expression level of Ldhc m RNA and protein in Plateau Pika skeletal muscle, and the levels of lactate dehydrogenase and lactic acid as well as the content of ATP were detected. The results showed that, compared to the control group, RNAi-LDHC group′s swimming time reduced by 12.45%（Fig. 1）. At the m RNA and protein levels, Ldhc gene expressions were reduced by 41.73% and 15.76%, respectively（Fig. 2）. LDH, LD and ATP were reduced by 23.98%, 51.08% and 19.29%, respectively（Fig. 3）. It is indicated that intraperitoneal injection of adenovirus p Multi RNAi-Ldhc can effectively silence Ldhc expression in Plateau Pika skeletal muscle.