Effects of Zhu Lian’s type Ⅱ inhibition needling on cerebral angiogenesis in rats with ischemic stroke based on the Ang-1/Tie-2 pathway

ObjectiveTo observe the effects of Zhu Lian’s type Ⅱ inhibition needling on neurological function and angiogenesis in rats with ischemic stroke （IS）， so as to explore its potential mechanisms.MethodsWistar rats were divided into sham operation group， model group， acupuncture group， and acupuncture+angiogenin （Ang）-2 group， with 12 rats in each group. A suture method was used to establish the IS model. The acupuncture group received Zhu Lian’s type Ⅱ inhibition needling at the “Guanyuan” （CV4）， “Guilai” （ST29）， “Zusanli” （ST36）， “Sanyinjiao” （SP6）， and “Fenglong” （ST40） acupoints for 30 min per session. Rats in the acupuncture+Ang-2 group were injected via the tail vein with Ang-2 recombinant protein （10 mg/kg） before Zhu Lian’s type Ⅱ inhibition needling intervention. All treatments were administered once daily for 7 consecutive days. Neurological deficit scores were assessed in rats of each group. TTC staining was employed to determine the percentage of cerebral infarction volume. HE staining was used to observe the pathological changes in the ischemic penumbra cortex. Immunohistochemistry was used to evaluate microvascular density in the ischemic penumbra cortex. Western blot was conducted to detect the expression levels of vascular endothelial growth factor （VEGF）， vascular endothelial growth factor receptor 2 （VEGFR-2）， Ang-1， Ang-2， tyrosine kinase 2 （Tie-2）， and phosphorylated Tie-2 （p-Tie-2） proteins in the ischemic penumbra cortex.ResultsCompared with the sham operation group， the model group showed elevated neurological deficit scores， cerebral infarction volume percentage， microvascular density in the ischemic penumbra cortex， and protein levels of VEGF， VEGFR-2， Ang-1， and p-Tie-2/Tie-2 ratio. Conversely， Ang-2 protein level was decreased （PPConclusionZhu Lian’s type Ⅱ inhibition needling can improve neurological function in IS rats， possibly by activating the Ang-1/Tie-2 pathway to promote angiogenesis.