Zhang Sai, Tian Yun-Na, Song Zheng-Yang, Wang Xiao-Ting, Wang Xin-Yu, Xu Jun-Peng, Yuan Lin-Bo, Wang Wan-Tie
Institute of Ischemia/Reperfusion Injury Research, Wenzhou Medical University, Wenzhou 325035, China.
Sheng Li Xue Bao. 2023 Aug 25;75(4):503-511.
In this study, we investigated the effects of Panax notoginseng saponins (PNS) on pulmonary vascular remodeling and ADAM10/Notch3 pathway in pulmonary arterial hypertension (PAH). PAH rat model was established, and male Sprague Dawley (SD) rats were randomly divided into control group, monocrotaline (MCT) group and MCT+PNS group, with 10 rats in each group. Rats in the control group were intraperitoneally injected with equal volume of normal saline. Rats in the MCT group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with the same volume of normal saline every day. Rats in the MCT+PNS group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with 50 mg/kg PNS every day. The modeling time of each group lasted for 21 days. After the model was established, the mean pulmonary artery pressure (mPAP) was measured by right heart catheterization technique, the right ventricular hypertrophy index (RVHI) was calculated, the microscopic morphology and changes of pulmonary vascular wall were observed by HE and Masson staining, and the expressions of ADAM10, Notch3, Hes-1, P27, PCNA, Caspase-3 proteins and mRNA in pulmonary vascular tissue of rats were detected by Western blot and qPCR. The expression and localization of Notch3 and α-SMA were detected by immunofluorescence staining. The protein expression of ADAM10 was detected by immunohistochemical staining. The results showed that compared with the control group, mPAP, RVHI, pulmonary vessels and collagen fibers in the MCT group were significantly increased, the expressions of ADAM10, Notch3, Hes-1, and PCNA protein and mRNA were significantly increased, while the expressions of P27 and Caspase-3 protein and mRNA were decreased significantly. Compared with the MCT group, mPAP and RVHI were significantly decreased, pulmonary vessels were significantly improved and collagen fibers were significantly reduced, the expressions of protein and mRNA of ADAM10, Notch3, Hes-1, and PCNA were decreased in MCT+PNS group, but the expressions of protein and mRNA of P27 and Caspase-3 were increased slightly. The results of immunofluorescence showed that Notch3 and α-SMA staining could overlap, which proved that Notch3 was expressed in smooth muscle cells. The expression of Notch3 in the MCT group was increased significantly compared with that in the control group, while PNS intervention decreased the expression of Notch3. Immunohistochemical staining showed that compared with the control group, the amount of ADAM10 in the MCT group was increased significantly, and the expression of ADAM10 in the MCT+PNS group was decreased compared with the MCT group. These results indicate that PNS can improve the PAH induced by MCT in rats by inhibiting ADAM10/Notch3 signaling pathway.
在本研究中，我们探究了三七总皂苷（PNS）对肺动脉高压（PAH）中肺血管重塑及ADAM10/Notch3信号通路的影响。建立PAH大鼠模型，将雄性Sprague Dawley（SD）大鼠随机分为对照组、野百合碱（MCT）组和MCT+PNS组，每组10只大鼠。对照组大鼠腹腔注射等体积的生理盐水。MCT组大鼠于第一天腹腔注射60 mg/kg MCT，随后每天注射等体积的生理盐水。MCT+PNS组大鼠于第一天腹腔注射60 mg/kg MCT，随后每天注射50 mg/kg PNS。每组的建模时间持续21天。模型建立后，采用右心导管技术测量平均肺动脉压（mPAP），计算右心室肥厚指数（RVHI），通过HE和Masson染色观察肺血管壁的微观形态及变化，采用蛋白质印迹法和qPCR检测大鼠肺血管组织中ADAM10、Notch3、Hes-1、P27、PCNA、Caspase-3蛋白及mRNA的表达。通过免疫荧光染色检测Notch3和α-SMA的表达及定位。采用免疫组织化学染色检测ADAM10的蛋白表达。结果显示，与对照组相比，MCT组的mPAP、RVHI、肺血管及胶原纤维显著增加，ADAM10、Notch3、Hes-1及PCNA蛋白和mRNA的表达显著增加，而P27和Caspase-3蛋白及mRNA的表达显著降低。与MCT组相比，MCT+PNS组的mPAP和RVHI显著降低，肺血管显著改善，胶原纤维显著减少，ADAM10、Notch3、Hes-1及PCNA的蛋白和mRNA表达降低，但P27和Caspase-3的蛋白和mRNA表达略有增加。免疫荧光结果显示，Notch3和α-SMA染色可重叠，证实Notch3在平滑肌细胞中表达。与对照组相比，MCT组中Notch3的表达显著增加，而PNS干预降低了Notch3的表达。免疫组织化学染色显示，与对照组相比，MCT组中ADAM10的量显著增加，与MCT组相比，MCT+PNS组中ADAM10的表达降低。这些结果表明，PNS可通过抑制ADAM10/Notch3信号通路改善MCT诱导的大鼠PAH。
