Shenjinhuoxue Mixture Attenuates Inflammation, Pain, and Cartilage Degeneration by Inhibiting TLR-4 and NF-κB Activation in Rats with Osteoarthritis: A Synergistic Combination of Multitarget Active Phytochemicals

Table 1

Pharmacological characteristic of the key active phytochemicals in SHM against OA.

Phytochemicals	Herbs	Molecular targets against OA	Effect on TLR-4	Other pharmacological activities	Pharmacokinetics	OB	DL	Toxicity
β-Sitosterol	Minister: RAS, RPA, MH, and RGM; assistant: VH and RD; guide: RC	Phosphorylation of NF-κB and the other components of the NF-κB pathway (↓) [29]	Protein expression (↓) [30]	SOD, CAT, and MDA (↓); GSH-PX, GSH, and Nrf2 (↑) [31]^a	/	36.91	0.75	No toxicity [32]
Oleanolic acid	Minister: RPA, RGM, and CPL; assistant: VH; guide: RC and RG	SIRT3 (↑) and NF-κB (↓) [33]; MMP-3 (↓); MMP-13, PGE2, IL-6, and caspase 9 (↓), PPARg and SOD2 (↑) [34]; RANKL-induced osteoclastogenesis (↓) [35]	Protein expression (↓) [36]	NF-κB, iNOS, TNF-α, IL-1β, and IL-6 (↓); MDA (↓); SOD, GSH-px, and Nrf2 (↑) [37]^a	: ; : ; : ; : ; CL/F: ; : (dose: 20 mg, oral, human) [38]	29.02	0.76	No influence on C57BL/6 mice at the concentrations below 90 mg/kg [39]
Licochalcone A	Guide: RG	Phosphorylation of NF-κB p65 and IκBα (↓), iNOS and COX-2 (↓); Wnt/β-catenin signaling (↓); Nrf2 and HO-1 (↑); [35, 36] RANKL-induced osteoclastogenesis (↓) [40]	Protein expression (↓) [41]	SIRT1/AMPK (↑) [42], NF-κB, AP-1, and, JNK (↓) [36]; Nrf2 signaling (↑); [41, 43]^a CYP3A4, CYP2C9, CYP1A2, CYP2C8, CYP2E1, CYP2D6, and P-gp (↓); BCRP and MRP2 (↑). [41–43]	/	40.79	0.29	No influence on HFF cell viability at the concentrations below 9 μg/mL (MTT cell viability assays) [44]
Quercetin	Minister: CPL and MH; guide: RC and RG	M2 polarization of synovial macrophages (↑) [45]; SIRT1/AMPK (↑); SOD and TIMP-1 (↑), MMP-13 (↓) [46]; TLR-4 and NF-κB (↓) [47]; RANKL-induced osteoclastogenesis (↓) [48]	Protein expression (↓) [49]	Akt/NF-κB signaling (↓); PI3K signaling, TLR4/MyD88/PI3K, JAK-STAT, NF-κB, p38 MAPK (↓); Nrf2 signaling, and AP-1 (↑) [50]^b; CYP2E1,CYP3A4,CYP2C19,MRP2,BCRP,P-gp, CYP1A2, and CYP2C8(↓); ABCA1, CYP1A1, and CYP2A6(↑) [51]	: 15.4 ng/ml; : 3 h; : 3.47 h; : 62.5 ng/h/mL; CL/F: 35300 L/h (dose: 500 mg, oral, human) [52]	46.43	0.28	>1500 mg/day with nephrotoxiciy [53]
Isorhamnetin	Assistant: VH	ROS production, RANKL-induced osteoclastogenesis, and MAPK/NF-κB/AKT signaling (↓) [54]; NF-κB and p65 (↓) [54]	Inhibiting the bond of LPS with TLR4 [55]	MAPK, NF-κB signaling (↓), PXR (↑); Nrf2 signaling (↑) [56]^b	: ; : ; : ; : ; : ; : ; CL/F: ; : ; : (dose:1.00 mg/kg, oral, rats) [57]	49.60	0.31	No toxicity [58]
Kaempferol	Minister: RPA; guide: RG	NF-κB (↓) [59]; MAPK-associated ERK and P38 signaling (↓) [60]	Protein expression (↓) [61]	TLR4/MyD88/NF-κB P65 signaling (↓); SOD, GPx, GCLC, and Nrf2 signaling (↑); UGT1A1, CYP3A4, P-gp, and CYP2E1 (↓) [62, 63]	: ; : ; CL: ; : (dose: 4 mg/kg, iv, rats) [64]	41.88	0.24	No data from in vivo studies evidencing these effects [65]
Morusin	Guide: RG	NF-κB signaling (↓) [66]	NONE	CYP3A4, CYP1A2, CYP2C9, CYP2E1, UGT1A6, UGT1A7, and UGT1A8 (↓) [67]	/	11.52	0.76	Unknown
Lupeol	Minister: MH; assistant: VH	RANKL, phosphorylation of MAPK, and NF-κB signaling (↓) [68]	Protein expression (↓) [69]	Phosphorylation of p38 MAPK, JNK, TLR4/MyD88/NF-κB P65 signaling, IRAK (↓), PI3K/Akt signaling, and caspase-3 activity; ROS (↓) and Nrf2 (↑) [70]^b	: ; : 6.444 h; : ; : ; CL/F: ; : ; (dose: 200 ng/kg, oral, CD-1 mice) [71]	12.12	0.78	No toxicity [72]
Pinocembrin	Guide: RG	NF-κB signaling (↓) [73]	Protein expression (↓) [74]	PI3K/Akt/NF-κB and MAPK signaling (↓), SIRT3 (↑); Erk1/2-Nrf2 (↑), SOD, MDA, and ROS (↓) [75]^a	: ; : ; CL/F: ; : (dose: 50 mg/kg, oral, SD rats) [76]	64.72	0.18	Unknown

Herbs of SHM—CPL: Carica papaya L.; CR: Cibot Rhizome; FK: Frankincense; MH: Myrrha; RAB: Radix Achyranthis Bidentatae; RAS: Radix Angelicae Sinensis; RC: Radix Cyathulae; RD: Radix Dipsaci; RG: Radix Glycyrrhizae; RGM: Radix Gentianae Macrophyllae; RPA: Radix Paeoniae Alba; VH: Visci Herba; AP-1: activator protein-1; AMPK: AMP-activated protein kinase; ABCG2: ATP-binding cassette transporter G2 and also known as breast cancer resistance protein (BCRP); Akt: protein kinase B; AUC_0-t: area under the concentration time curve from zero to time; COX: cyclooxygenase; CAT: catalase; CL: clearance; CL/F: apparent clearance; C_max: the maximum plasma concentrations; DL: druglikeness; ERK: extracellular-regulated kinase; GSH: glutathione; GSH-Px: glutathione peroxidase; HO-1: hemeoxygenase-1; IL: interleukin; IRAK: interleukin-1 receptor-associated kinase; IκBα: nuclear factor of kappa light polypeptide gene enhancer in B-cell inhibitor, alpha; IRF5: interferon regulatory factor 5; JAK-STAT: Janus kinase/signal transduction and activator of transcription; JNK: Jun N-terminal kinase; MAPK: mitogen-activated protein kinase; MDA: malondialdehyde; MMP: matrix metalloproteinase; MRP2: multidrug resistance protein 2; MyD88: myeloid differentiation primary response gene 88; NF-κB: nuclear factor kappa B; Nrf2: nuclear factor- (erythroid derived 2-) like 2; OB: oral bioavailability; PI3K: phosphatidylinositol-3-kinase; PPAR: peroxisome proliferator-activated receptor; P-gp: P-glycoprotein 1; RANKL: receptor activator of the NF-κB ligand; ROS: reactive oxygen species; SIRT 3: sirtuin 3; SOD: superoxide dismutase; TLR4: Toll-like receptor 4; : time taken to reach the ; : half-life; : the distribution half-life; : the elimination half-life; : the absorption half-life; TNF-α: tumor necrosis factor-α; UGT: UDP-glucuronosyltransferase; : volume of distribution; : apparent volume of distribution; : apparent volume of distribution to the central compartment. ^aHepatoprotection; ^bhepatoprotection and renoprotection.