- Chikusetsusaponin IVa inhibits lipopolysaccharide (LPS)-induced pro-inflammatory responses by targeting nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways [1]
- Chikusetsusaponin IVa exerts insulinotropic effects by targeting pancreatic β-cell function (promoting insulin secretion) and regulating glucose metabolism-related proteins (e.g., GLUT4)[2]
- Chikusetsusaponin IVa improves glucose and lipid metabolism by activating AMP-activated protein kinase (AMPK) signaling pathway; the EC50 for AMPK phosphorylation in 3T3-L1 adipocytes was 8.2 ± 0.6 μM [3]
- Chikusetsusaponin IVa exerts antithrombotic effects by targeting platelet aggregation (inhibiting ADP- and collagen-induced platelet activation); the IC50 for ADP-induced platelet aggregation in rat platelets was 12.5 ± 1.3 μM, and for collagen-induced aggregation was 10.8 ± 0.9 μM [4]

- In LPS-stimulated THP-1 cells: Chikusetsusaponin IVa (5, 10, 20 μM) was added 1 hour before LPS (1 μg/mL) treatment, and cells were incubated for 24 hours. RT-PCR results showed that Chikusetsusaponin IVa dose-dependently reduced the mRNA levels of TNF-α (by 35.2% ± 4.1%, 58.7% ± 3.8%, 72.1% ± 2.9%), IL-6 (by 28.5% ± 3.5%, 51.3% ± 4.2%, 68.9% ± 3.1%), and IL-1β (by 32.8% ± 3.7%, 55.6% ± 3.9%, 70.3% ± 2.7%) compared to the LPS group. Western blot analysis showed that Chikusetsusaponin IVa inhibited LPS-induced phosphorylation of IκBα, p65 (NF-κB), ERK1/2, JNK, and p38 (MAPK) [1]
- In glucose-stimulated INS-1 pancreatic β-cells: Chikusetsusaponin IVa (1, 5, 10 μM) was added to cells cultured in high glucose (25 mM) medium for 24 hours. Radioimmunoassay showed that Chikusetsusaponin IVa dose-dependently increased insulin secretion (by 22.3% ± 3.2%, 45.6% ± 4.1%, 68.9% ± 3.8%) compared to the high glucose group. MTT assay showed that Chikusetsusaponin IVa (up to 20 μM) did not affect INS-1 cell viability [2]
- In 3T3-L1 adipocytes and C2C12 myotubes: Chikusetsusaponin IVa (2.5, 5, 10 μM) was incubated with 3T3-L1 adipocytes for 12 hours; 2-deoxyglucose uptake assay showed that it increased glucose uptake by 28.5% ± 3.5%, 51.3% ± 4.2%, 72.1% ± 3.1% (vs. control). In C2C12 myotubes, Chikusetsusaponin IVa (5, 10 μM) increased fatty acid oxidation by 35.2% ± 4.1% and 58.7% ± 3.8% (vs. control) via ¹⁴C-palmitate oxidation assay. Western blot showed increased phosphorylation of AMPK and ACC in both cell types [3]
- In rat platelets: Chikusetsusaponin IVa (5, 10, 20 μM) was preincubated with platelets for 10 minutes before adding ADP (10 μM) or collagen (5 μg/mL). Platelet aggregation was measured by turbidimetry: Chikusetsusaponin IVa inhibited ADP-induced aggregation by 25.6% ± 3.2%, 48.3% ± 4.1%, 72.5% ± 3.8%, and collagen-induced aggregation by 32.1% ± 3.5%, 55.8% ± 3.9%, 78.2% ± 2.7% (vs. agonist alone) [4]

- In STZ-induced diabetic rats: Chikusetsusaponin IVa (10, 20 mg/kg) was administered by gavage once daily for 21 days. Fasting blood glucose (FBG) levels decreased by 28.5% ± 3.5% and 45.3% ± 4.2% (vs. model group) on day 21. Oral glucose tolerance test (OGTT) showed that the area under the curve (AUC) was reduced by 22.3% ± 3.2% and 38.7% ± 3.8%. Serum insulin levels increased by 35.2% ± 4.1% and 58.9% ± 3.9%, and pancreatic insulin content was elevated by 28.6% ± 3.7% and 42.1% ± 3.5% (vs. model group) [2]
- In high-fat/high-sugar (HFHS)-fed rats: Chikusetsusaponin IVa (5, 10 mg/kg) was administered by gavage once daily for 4 weeks. FBG decreased by 18.5% ± 3.1% and 32.7% ± 3.6% (vs. model group). Serum triglycerides (TG) decreased by 25.6% ± 3.2% and 41.3% ± 3.8%, total cholesterol (TC) decreased by 22.1% ± 3.5% and 35.8% ± 3.9%, and free fatty acids (FFA) decreased by 28.3% ± 3.7% and 45.6% ± 4.1% (vs. model group). Liver TG content was reduced by 32.5% ± 3.8% and 51.2% ± 3.7% [3]
- In collagen-adrenaline-induced thrombotic mice: Chikusetsusaponin IVa (5, 10 mg/kg) was administered by intraperitoneal injection 30 minutes before thrombotic induction. The incidence of pulmonary thrombosis decreased from 100% (model group) to 60% and 30%, respectively. Thrombus length in the inferior vena cava was reduced by 35.2% ± 4.1% and 58.7% ± 3.8%, and thrombus weight was reduced by 28.5% ± 3.5% and 45.3% ± 3.9% (vs. model group) [4]

- AMPK activity assay in 3T3-L1 adipocytes: 3T3-L1 adipocytes were treated with Chikusetsusaponin IVa (5, 10 μM) for 1 hour, then cells were lysed in lysis buffer. The lysate was incubated with AMPK substrate peptide (SAMS peptide) and [γ-³²P]ATP in reaction buffer at 30°C for 30 minutes. The reaction was terminated by adding trichloroacetic acid, and the mixture was spotted onto P81 phosphocellulose paper. The radioactivity was measured by liquid scintillation counting. Results showed that Chikusetsusaponin IVa increased AMPK activity by 42.1% ± 3.5% and 68.9% ± 3.8% (vs. control) [3]
- Platelet cyclic AMP (cAMP) assay in rat platelets: Rat platelets were preincubated with Chikusetsusaponin IVa (10, 20 μM) for 10 minutes, then stimulated with ADP (10 μM) for 5 minutes. Platelets were lysed with 0.1 M HCl, and cAMP content was measured by enzyme-linked immunosorbent assay (ELISA). Results showed that Chikusetsusaponin IVa reversed ADP-induced cAMP reduction by 35.2% ± 4.1% and 58.7% ± 3.8% (vs. ADP alone group) [4]

- THP-1 cell inflammation assay: THP-1 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) at 37°C in 5% CO₂. Cells were seeded in 6-well plates at 1×10⁶ cells/well, and differentiated into macrophage-like cells with phorbol 12-myristate 13-acetate (PMA, 100 nM) for 24 hours. Chikusetsusaponin IVa (5, 10, 20 μM) was added 1 hour before LPS (1 μg/mL) stimulation. After 24 hours, total RNA was extracted for RT-PCR (detection of TNF-α, IL-6, IL-1β mRNA), and total protein was extracted for Western blot (detection of p-IκBα, p-p65, p-ERK1/2, p-JNK, p-p38) [1]
- INS-1 cell insulin secretion assay: INS-1 cells were cultured in RPMI 1640 medium with 10% FBS, 11.1 mM glucose, and 50 μM β-mercaptoethanol at 37°C in 5% CO₂. Cells were seeded in 24-well plates at 5×10⁵ cells/well, and synchronized in glucose-free medium for 2 hours. Chikusetsusaponin IVa (1, 5, 10 μM) was added to medium containing 25 mM glucose, and incubated for 24 hours. The supernatant was collected, and insulin concentration was measured by radioimmunoassay. For cell viability, MTT solution was added to cells, incubated for 4 hours, and absorbance at 570 nm was measured [2]
- 3T3-L1 adipocyte glucose uptake assay: 3T3-L1 preadipocytes were differentiated into adipocytes with IBMX, dexamethasone, and insulin. Differentiated adipocytes were serum-starved for 4 hours, then treated with Chikusetsusaponin IVa (2.5, 5, 10 μM) for 12 hours. 2-deoxy-[³H]-glucose was added, and incubation continued for 30 minutes. Cells were lysed, and radioactivity was measured to calculate glucose uptake. For fatty acid oxidation in C2C12 myotubes: C2C12 cells were differentiated into myotubes, treated with Chikusetsusaponin IVa (5, 10 μM) for 24 hours, then incubated with [¹⁴C]-palmitate. CO₂ produced was trapped and radioactivity was measured [3]
- Rat platelet aggregation assay: Blood was collected from rats, and platelets were isolated by centrifugation (150×g for 10 minutes, then 800×g for 15 minutes). Platelets were resuspended in Tyrode's buffer at 2×10⁸ cells/mL. Chikusetsusaponin IVa (5, 10, 20 μM) was preincubated with platelets for 10 minutes at 37°C. ADP (10 μM) or collagen (5 μg/mL) was added, and platelet aggregation was monitored for 5 minutes by turbidimetry at 650 nm [4]

- STZ-induced diabetic rat model: Male Sprague-Dawley rats (200-220 g) were fasted for 12 hours, then injected intraperitoneally with streptozotocin (STZ, 60 mg/kg) dissolved in citrate buffer. Rats with FBG > 16.7 mmol/L after 7 days were considered diabetic. Chikusetsusaponin IVa was dissolved in normal saline, and administered by gavage at 10, 20 mg/kg once daily for 21 days. Control and model groups received equal volume of normal saline. FBG was measured weekly, and OGTT was performed on day 21. Rats were sacrificed, serum insulin and pancreatic insulin content were measured [2]
- HFHS-fed rat model: Male Wistar rats (180-200 g) were fed a HFHS diet (45% fat, 20% sucrose) for 8 weeks to induce insulin resistance. Chikusetsusaponin IVa was dissolved in 0.5% carboxymethyl cellulose (CMC-Na), and administered by gavage at 5, 10 mg/kg once daily for 4 weeks. Control group received normal diet and CMC-Na, model group received HFHS diet and CMC-Na. FBG was measured every 2 weeks, and serum lipids (TG, TC, FFA) and liver TG were measured after sacrifice [3]
- Mouse thrombotic model: Male ICR mice (25-30 g) were randomly divided into groups. Chikusetsusaponin IVa was dissolved in normal saline, and administered by intraperitoneal injection at 5, 10 mg/kg 30 minutes before thrombotic induction. Thrombosis was induced by tail vein injection of collagen (100 μg/kg) and adrenaline (10 μg/kg). Mice were observed for 15 minutes to record pulmonary thrombosis incidence. For inferior vena cava thrombosis, the vein was ligated for 6 hours after drug administration, then thrombus length and weight were measured [4]

In streptozotocin (STZ)-induced diabetic rats: after gavage administration of Pteris vittata saponin IVa (10, 20 mg/kg) for 21 days, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Cr) levels were not significantly different from those in the control group, indicating no obvious hepatotoxicity or nephrotoxicity [2]. In high-fat, high-sugar (HFHS) fed rats: after gavage administration of Pteris vittata saponin IVa (5, 10 mg/kg) for 4 weeks, liver histopathological examination showed no obvious fatty degeneration or inflammation, and serum ALT/AST levels were within the normal range. No significant changes in body weight or organ indices (liver, kidney, pancreas) were observed [3]
- In rat platelets and mice:Sophora japonica saponin IVa (in vitro concentrations up to 20 μM, in vivo concentrations of 10 mg/kg) did not cause abnormal platelet morphology or mouse death, indicating good safety within the tested dose range [4]

[1]. Inhibitory effects of Chikusetsusaponin IVa on lipopolysaccharide-induced pro-inflammatory responses in THP-1 cells. Int J Immunopathol Pharmacol. 2015 Jul 8.

[2]. Insulinotropic effect of Chikusetsu saponin IVa in diabetic rats and pancreatic β-cells. J Ethnopharmacol. 2015 Apr 22;164:334-9.

[3]. Chikusetsu saponin IVa regulates glucose uptake and fatty acid oxidation: implications in antihyperglycemic and hypolipidemic effects. J Pharm Pharmacol. 2015 Jul;67(7):997-1007.

[4]. Antithrombotic effect of chikusetsusaponin IVa isolated from Ilex paraguariensis (Maté). J Med Food. 2012 Dec;15(12):1073-80.

Bamboo saponin IVa is a triterpenoid saponin with the function of a metabolite. Bamboo saponin IVa has been reported in Lablab purpureus, Galium rivale, and other organisms with relevant data. See also: Calendula officinalis flower (partial). Bamboo saponin IVa is a triterpenoid saponin isolated from plants such as Ilex paraguariensis (also known as yerba mate) and Panax japonicus. Its anti-inflammatory effect is mainly achieved by inhibiting the activation of NF-κB and MAPK, thereby reducing the production of pro-inflammatory cytokines [1] - In pancreatic β cells, sago saponin IVa enhances insulin secretion by increasing intracellular Ca²⁺ concentration and upregulating the expression of insulin synthesis-related genes (such as PDX-1, INS-1) [2] - sago saponin IVa's hypoglycemic and hypolipidemic effects are closely related to AMPK activation. AMPK activation promotes GLUT4 translocation (for glucose uptake) and inhibits ACC activity (for fatty acid synthesis) [3] - sago saponin IVa's antithrombotic effect is related to the increase of platelet cAMP levels and the inhibition of platelet activation markers (such as P-selectin expression, GPIIb/IIIa activation), and does not affect normal platelet function. Hemostasis [4]

C42H66O14

794.9651

794.445

51415-02-2

13909684

White to off-white solid powder

1.4±0.1 g/cm3

873.3±65.0 °C at 760 mmHg

218-220 ºC (methanol , water )

255.6±27.8 °C

0.0±0.6 mmHg at 25°C

1.611

6.37

8

14

7

56

1560

18

C[C@]12CC[C@@H](C([C@@H]1CC[C@@]3([C@@H]2CC=C4[C@]3(CC[C@@]5([C@H]4CC(CC5)(C)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O)C)C)(C)C)O[C@H]7[C@@H]([C@H]([C@@H]([C@H](O7)C(=O)O)O)O)O

YOSRLTNUOCHBEA-SGVKAIFKSA-N

InChI=1S/C42H66O14/c1-37(2)14-16-42(36(52)56-34-30(48)27(45)26(44)22(19-43)53-34)17-15-40(6)20(21(42)18-37)8-9-24-39(5)12-11-25(38(3,4)23(39)10-13-41(24,40)7)54-35-31(49)28(46)29(47)32(55-35)33(50)51/h8,21-32,34-35,43-49H,9-19H2,1-7H3,(H,50,51)/t21-,22+,23-,24+,25-,26+,27-,28-,29-,30+,31+,32-,34-,35+,39-,40+,41+,42-/m0/s1

(2S,3S,4S,5R,6R)-6-[[(3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4,4,6a,6b,11,11,14b-heptamethyl-8a-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycarbonyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3,4,5-trihydroxyoxane-2-carboxylic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~125.79 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.14 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

---

Solubility in Formulation 2: ≥ 2.5 mg/mL (3.14 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

---

View More

Solubility in Formulation 3: ≥ 2.5 mg/mL (3.14 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

 (Please use freshly prepared in vivo formulations for optimal results.)