Astragaloside IV (AS-IV), an active component isolated from Astragalus membranaceus, suppresses the activation of ERK1/2 and JNK, and downregulates matrix metalloproteases (MMP)-2, (MMP)-9 in MDA-MB-231 breast cancer cells. Astragaloside IV is a bioactive saponin first isolated from the dried plant roots of the genus Astragalus, which is used in traditional Chinese medicine.1 It dose-dependently inhibits human adenovirus type 3 (HAdV-3) in A549 cells (IC50 = 23 μM; LC50 = 865 μM).It inhibits replication of HAdV-3 and decreases HAdV-3-induced apoptosis. It has diverse protective effects for the cardiovascular, immune, digestive, and nervous systems. In particular, it reduces myocardial infarct size in dogs when administered prior to coronary ligation and reduces reperfusion arrhythmias in isolated rat hearts.

Astragaloside IV对MDA-MB-231乳腺癌细胞的活性和侵袭性产生抑制作用，能够压抑丝裂原活化蛋白激酶（MAPK）家族成员ERK1/2和JNK的激活，并下调金属蛋白酶（MMP）-2和-9的表达。在NSCLC细胞生长抑制方面，Astragaloside IV（10、20、40 ng/mL）显示出明显的作用，而低浓度Astragaloside IV（1、2.5、5 ng/mL）对细胞活性无明显细胞毒性。此外，Astragaloside IV与化疗化合物顺铂联合治疗显著提高了NSCLC细胞对化疗化合物的敏感性。在分子水平上，Astragaloside IV与顺铂联合使用显著抑制了B7-H3的mRNA和蛋白水平的表达。

在小鼠模型中，高剂量的Astragaloside IV组在48小时存活率上显示出显著增加[60%(9/15)对比13.3%(2/15), P<0.05]，血清ALT和AST水平显著降低(P<0.01)，肝脏组织病理学指数及肝细胞凋亡程度显著降低(P<0.01)，以及肝匀浆中MDA含量显著减少(P<0.01)，SOD活性显著增加。Astragaloside IV（10, 20 mg/kg, p.o.）显著地预防了短暂脑缺血及再灌注所诱导的认知缺陷。与模型组相比，Astragaloside IV（10 mg/kg）和Astragaloside IV（20 mg/kg）能显著降低这些细胞因子的水平。Astragaloside IV明显抑制了TLR4及其下游蛋白的水平，表明MyD88依赖性和非依赖性途径在Astragaloside IV的抗炎作用中发挥了重要作用。此外，Astragaloside IV减少了NLRP3和活化的caspase-1的表达，同时也降低了Iba1蛋白的表达。

MDA-MB-231 cells treated as indicated or tumor tissues are harvested and lysed in Mg2+ lysis buffer containing 50 mM Tris (pH 7.5), 10 mM MgCl2, 0.5 M NaCl, and protease inhibitor cocktail. Equal amounts of lysates are incubated with PAK-PBD beads at 4°C for 1 h. PAK-PBD beads are pelleted by centrifugation and washed with ish buffer containing 25 mM Tris (pH 7.5), 30 mM MgCl2, 40 mM NaCl. Active Rac1 is detected by western blotting.

CCK-8 assay is adopted to determine cell viability. cultured NSCLC cells are seeded into 96-well plates at the density of 4×104 (cells/well). Then 10 μL?well CCK8 solution is added and incubated in dark at 37°C for another 2 h. The absorbance is determined with the wavelength of 490 nm.

Transient cerebral ischemia and reperfusion is prepared by BCCAO. Mice are randomly divided into the Sham, Model, Astragaloside IV (10 mg/kg) and Astragaloside IV (20 mg/kg) treatment groups. The Astragaloside IV treatment groups are intragastrically administered 7 days before the surgery and terminated on the day of sacrifice. On the day of the surgery, Astragaloside IV is administrated 2 h prior to ischemia. The Sham-operated and Model groups are treated with distilled water. After the mice are anesthetized with an intraperitoneal injection of chloral hydrate (350 mg/kg), the bilateral common carotid arteries are exposed and carefully separated with a small ventral neck incision and occluded twice (20 min each) with ligated surgical silk as described previously with minor modifications. There is a 10 min reperfusion period between the two occlusion periods (ischemia 20 min ? reperfusion 10 min ? ischemia 20 min). Sham-operated mice are subjected to the same surgical operation without the surgical silk ligation. Mouse body temperature is maintained at 37±0.5°C during the surgery with heating equipment until recovery from the anesthesia.