Br J Pharmacol: Amitriptyline blocks TLR4 and IL1R-mediated innate immune response: preclinical and clinical evidence for OA and gout

In modern society, musculoskeletal diseases are increasing day by day, which adversely affects our society

Experimental method: Through electronic docking analysis, RT-PCR, siRNA, ELISA, proteomics, and clinical data mining of drug use, the clinical significance of amitriptyline in blocking TLR4-mediated IIRs in hOCs was explored

Experimental design: Four options were determined: control group (untreated and unstimulated cells), drugs (amitriptyline [0.

In the specific experiment process, CLI-095 [1µM] or NLRP3 siRNA [15 nm] was used instead of amitriptyline

In terms of sample size, the availability of primary cells is very small, but we have established a group (n) consisting of 6 independent biological copies, each copy has 3 technology-dependent copies, suitable for the determination of all cell types

Results: Amitriptyline can bind to TLR4 but not IL1 receptor (IL1R)

Figure 1 The effect of amitriptyline on TLR4-mediated innate immune response factors

Figure 1 The effect of amitriptyline on TLR4-mediated innate immune response factors

Figure 2 Amitriptyline reverses TLR4-mediated proteomics changes

Figure 2 Amitriptyline reverses TLR4-mediated proteomics changes

Figure 3 The effect of amitriptyline on Il1R-mediated innate immune response factors.

Figure 3 The effect of amitriptyline on Il1R-mediated innate immune response factors.

Figure 4 Amitriptyline reverses IL1R-mediated proteomics changes

Figure 4 Amitriptyline reverses IL1R-mediated proteomics changes

Figure 5 Amitriptyline blocks NLRP3 inflammasome

Figure 5 Amitriptyline blocks NLRP3 inflammasome
.
All experiments are independent, standardized by the control group, and expressed as mean ± SEM
.
AC: Before TLR4 activation (LPS [100ng/ml]) or IL1R activation (IL1β [0.
1ng/ml]), human OA chondrocytes (hOC) were treated with cli095[1µM] and treated with AT[1µM] for 24h , To detect gene expression of NLRP3, IL1β mRNA (n=6) and IL1β secretion level (n=3)
.
DG: Before TLR4 activation (LPS [100ng/ml]) or IL1R activation (IL1β [0.
1ng/ml]), combined treatment of ATDC5 chondrocytes with cli095[1µM] or NLRP3 siRNA [15nM] for 24h (n=6), Detect the gene expression of NLRP3 and IL1β mRNA (RT-PCR)
.
HK: Before TLR4 activation (LPS [100ng/ml]) or IL1R activation (IL1β [0.
1ng/ml]) and AT[1µM] or NLRP3 siRNA [15nM] are combined for 24h (n=6), use cli095[1µM ][1µM] or NLRP3 siRNA [15nM] treated human SW982 synovial cells for 3h, and detected the gene expression of NLRP3 and IL1β mRNA (RT-PCR)
.
LN: Treat human SW982 synovial cells with OA (n=6) or gout (n=4) synovial fluid (10% v/v) combined with AT (1µM) for 24 hours, and detect MTT cell viability and NLRP3 and IL1β mRNA genes Expression (RTPCR)
.

Figure 6 Amitriptyline reduces the need for colchicine to treat gout.
107.
172 elderly people (>65 years old) from the health care area of ​​Santiago de Compostela (Spain) were subjected to 5 years of clinical data mining
.
A: The Fisher test compares whether the elderly (>65 years old) need colchicine and amitriptyline
.
B: The Spearman related test was used to compare the dosage of colchicine (dose: up to 0.
5 mg; the exact average dose is 0.
66 mg) and amitriptyline (the most commonly used dose is 25 mg; the kit for individual consumption by elderly (>65 years) patients)
.
C: The effect of amitriptyline on the activation and inhibition of TLR4 and IL1R crosstalk signals, including the NLRP3/IL1B axis, NFκβ and AP1 myd88-dependent pathways and hOC
.

Figure 6 Amitriptyline reduces the need for colchicine to treat gout.
107.
172 elderly people (>65 years old) from the health care area of ​​Santiago de Compostela (Spain) were subjected to 5 years of clinical data mining
.
A: The Fisher test compares whether the elderly (>65 years old) need colchicine and amitriptyline
.
B: The Spearman related test was used to compare the dosage of colchicine (dose: up to 0.
5 mg; the exact average dose is 0.
66 mg) and amitriptyline (the most commonly used dose is 25 mg; the kit for individual consumption by elderly (>65 years) patients)
.
C: The effect of amitriptyline on the activation and inhibition of TLR4 and IL1R crosstalk signals, including the NLRP3/IL1B axis, NFκβ and AP1 myd88-dependent pathways and hOC
.

Conclusion: We proved that amitriptyline can block IIRs that depend on TLR4, IL1R and NLRP3
.
This activity and clinical data indicate that amitriptyline can be used for systemic or local IIR treatment of different joint pathologies
.

We proved that amitriptyline can block IIRs that depend on TLR4, IL1R and NLRP3
.
This activity and clinical data indicate that amitriptyline can be used for systemic or local IIR treatment of different joint pathologies
.

Original source:

Franco-Trepat E, Alonso-Pérez A, Guillán-Fresco M, et al.
Amitriptyline blocks innate immune responses mediated by TLR4 & IL1R: preclinical and clinical evidence in OA and gout .
Br J Pharmacol 2021 Oct 13

Amitriptyline blocks innate immune responses mediated by TLR4 & IL1R: preclinical and clinical evidence in OA and gout

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