Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: Experimental designs and animal groups. Experiment 1: Changes in mechanical allodynia; the expression of DKK3, Kremen-1, DVL-1, and ASK-1/JNK/p38 MAPK pathway after SNI in rats. Experiment 2: The effects of exogenous rDKK3 administration on mechanical allodynia, microglia polarization and neuroinflammation induced by neuropathic pain. Experiment 3: Kremen-1 siRNA or DVL-1 siRNA abolished the effects of rDKK3 on mechanical allodynia, microglia polarization and neuroinflammation caused by neuropathic pain

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Expressing

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: Analgesic effect of rDKK3 on mechanical allodynia in neuropathic pain rats. a , b A single dose of rDKK3 (30 and 50 μg) markedly increased the ipsilateral PWT in SNI rats, beginning at 2 h and lasted for at least 4 h, and didn’t affect the contralateral PWT ( *** p < 0.001, **** p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group). c , d Repetitive injections of rDKK3 (30 μg, i.t.) considerably reversed established mechanical allodynia in the ipsilateral side of SNI rats, and didn’t affect the contralateral PWT ( **** p < 0.0001 compared with Sham + Vehicle group, # p < 0.05, ## p < 0.01 compared with SNI + Vehicle group, n = 6 in each group). e , f The ipsilateral PWT was significantly increased from day 3 to day 8 in rDKK3-treated SNI rats compared with vehicle-treated SNI rats ( **** p < 0.0001 compared with Sham + Vehicle group, # p < 0.05, ## p < 0.01 compared with SNI + Vehicle group, n = 6 in each group). And the contralateral PWT in sham and SNI rats treated with Vehicle or rDKK3 had no significant change during the observation period ( p > 0.05). g Horizontal movement traces in the OFT of sham and SNI rats treated with Vehicle or rDKK3. h , i The OFT result showed that there was no significant difference in the total distance and average speed between the Sham + Vehicle group, Sham + rDKK3 group, SNI + Vehicle group, and SNI + rDKK3 group ( p > 0.05)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: rDKK3 inhibited the activation of ASK1/JNK/p-38 signaling pathway in rats with neuropathic pain. a rDKK3 could increase the reduced protein expression level of DKK3 in neuropathic pain rats ( ** p < 0.01 compared with Sham + Vehicle group, ### p < 0.001 compared with SNI + Vehicle group, n = 6 in each group). b , c Western blot results indicated that the protein level of ASK1 in sham and SNI rats treated with Vehicle or rDKK3 had no significant change, while rDKK3 down-regulated the elevated protein expression level of p-ASK1 induced by neuropathic pain in the spinal cord ( ** p < 0.01 compared with Sham + Vehicle group, ### p < 0.001 compared with SNI + Vehicle group, n = 6 in each group). d , e Western blot results indicated that the protein level of JNK in sham and SNI rats applied with Vehicle or rDKK3 had no significant change, while rDKK3 down-regulated the increased protein expression level of p-JNK caused by neuropathic pain in the spinal cord ( **** p < 0.0001 compared with Sham + Vehicle group, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group). f , g Western blot result indicated that the protein level of p38 in sham and SNI rats administrated with Vehicle or rDKK3 had no significant change, while rDKK3 alleviated the increased protein expression level of p-p38 in the spinal cord of rats with neuropathic pain ( **** p < 0.0001 compared with Sham + Vehicle group, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: rDKK3 promoted the switch of microglia from M1 type to M2 type in rats with neuropathic pain. a – c Western blot and immunofluorescence results showed that rDKK3 ameliorated the up-regulated protein expression level of Iba-1 in the spinal cord and suppressed the activated microglia caused by neuropathic pain ( **** p < 0.0001 compared with Sham + Vehicle group, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group). d – f Western blot results indicated that treatment with rDKK3 attenuated the increased protein expression level of CD16, CD86 and iNOS in the spinal cord induced by neuropathic pain ( **** p < 0.0001 compared with Sham + Vehicle group, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group). g – i Western blot result indicated that application with rDKK3 up-regulated the decreased protein expression level of Arg1, CD206 and IL-10 in the spinal cord caused by neuropathic pain ( * p < 0.05, ** p < 0.01, *** p < 0.001 compared with Sham + Vehicle group, # p < 0.05, ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Western Blot, Immunofluorescence, Expressing

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: The improved mechanical allodynia was reversed by intrathecal injection with Kremen-1 siRNA or DVL-1 siRNA. a Western blot results showed that Kremen-1 siRNA effective down-regulated the protein level of Kremen-1 ( **** p < 0.0001 compared with scramble siRNA group, n = 6 in each group). b Western blot results showed that DVL-1 siRNA successful down-regulated the protein level of DVL-1 ( **** p < 0.0001 compared with scramble siRNA group, n = 6 in each group). c , d rDKK3 alleviated the decreased ipsilateral PWT induced by neuropathic pain, while application with Kremen-1 siRNA or DVL-1 siRNA both could abolish the analgesic effect of rDKK3 ( **** p < 0.0001 compared with Sham group, ### p < 0.001, #### p < 0.001 compared with SNI + Vehicle group, & p < 0.05, && p < 0.01, &&& p < 0.001 &&&& p < 0.0001 compared with SNI + rDKK3 group, n = 6 in each group). In contrast, the contralateral PWT in the six groups had no significant change. (e) Horizontal movement traces in the OFT of the Sham group, SNI + Vehicle group, SNI + rDKK3 group, SNI + rDKK3 + scramble siRNA group, SNI + rDKK3 + Kremen-1 siRNA group, and SNI + rDKK3 + DVL-1 siRNA group. f , g The OFT result showed that there was no significant difference in the total distance and average speed between the Sham group, SNI + Vehicle group, SNI + rDKK3 group, SNI + rDKK3 + scramble siRNA group, SNI + rDKK3 + Kremen-1 siRNA group, and SNI + rDKK3 + DVL-1 siRNA group ( p > 0.05)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Injection, Western Blot

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: rDKK3 inhibited ASK1/JNK/p38 MAPK signaling pathway via Kremen-1 and DVL-1. a , b The expression level of ASK1 in the six groups had no significant change. rDKK3 reduced the increased p-ASK1 in neuropathic pain rats, while Kremen-1 siRNA or DVL-1 siRNA both reversed the down-regulated p-ASK1 caused by rDKK3 in neuropathic pain rats ( *** p < 0.001 compared with Sham group, ## p < 0.01 compared with SNI + Vehicle group, && p < 0.01 compared with SNI + rDKK3 group, n = 6 in each group). c , d The expression level of JNK in the six groups had no significant change. rDKK3 reduced the increased p-JNK in rats with SNI, while Kremen-1 siRNA or DVL-1 siRNA both up-regulated the decreased p-JNK caused by rDKK3 in rats with neuropathic pain ( **** p < 0.0001 compared with Sham group, #### p < 0.0001 compared with SNI + Vehicle group, &&&& p < 0.0001 compared with SNI + rDKK3 group, n = 6 in each group). e , f The expression level of p38 in the six groups had no significant change. rDKK3 dampened the increased p-p38 in neuropathic pain rats, while Kremen-1 siRNA or DVL-1 siRNA both elevated the decreased p-p38 caused by rDKK3 in neuropathic pain rats ( * p < 0.05 compared with Sham group, # p < 0.05 compared with SNI + Vehicle gro up, & p < 0.05, && p < 0.01 compared with SNI + rDKK3 group, n = 6 in each group)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Expressing

Journal: Journal of Neuroinflammation

Article Title: DKK3 ameliorates neuropathic pain via inhibiting ASK-1/JNK/p-38-mediated microglia polarization and neuroinflammation

doi: 10.1186/s12974-022-02495-x

Figure Lengend Snippet: rDKK3 promoted the switch of microglia from M1 type to M2 type in neuropathic pain rats via Kremen-1 and DVL-1. a – c Western blot and immunofluorescence results showed that rDKK3 ameliorated the up-regulated protein expression level of Iba-1 in the spinal cord and suppressed the activated microglia caused by neuropathic pain, while treatment with Kremen-1 siRNA or DVL-1 siRNA abolished the effect of rDKK3 on microglia activation ( **** p < 0.0001 compared with Sham group, #### p < 0.0001 compared with SNI + Vehicle group, &&&& p < 0.0001 compared with SNI + rDKK3 group, n = 6 in each group). d – f Western blot results indicated that treatment with rDKK3 attenuated the increased protein expression level of CD16, CD86 and iNOS in the spinal cord induced by neuropathic pain, while Kremen-1 siRNA or DVL-1 siRNA abolished the effect of rDKK3 on M1 type biomarkers ( ** p < 0.01, *** p < 0.001, **** p < 0.0001 compared with Sham group, ## p < 0.01, ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group, && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 compared with SNI + rDKK3 group, n = 6 in each group). g – i Western blot results indicated that application with rDKK3 up-regulated the decreased protein expression level of Arg1, CD206 and IL-10 in the spinal cord caused by neuropathic pain, while administration with Kremen-1 siRNA or DVL-1 siRNA attenuated the effect of rDKK3 on M2 type biomarkers (** p < 0.01, **** p < 0.0001 compared with Sham group, # p < 0.05, #### p < 0.0001 compared with SNI + Vehicle group, && p < 0.01, &&&& p < 0.0001 compared with SNI + rDKK3 group, n = 6 in each group)

Article Snippet: rDKK3 (SRP6268, Sigma-Aldrich, MO, USA) was dissolved in saline.

Techniques: Western Blot, Immunofluorescence, Expressing, Activation Assay

Journal: American Journal of Translational Research

Article Title: Use of tumor suppressor genes of naked mole rats for human cancer treatment

doi:

Figure Lengend Snippet: Phylogenetic gain/loss trees of DKK3 gene (A) and PDCD5 gene (B) were created in www.ensembl.com. The trees showed the number of DKK3 and PDCD5 paralogues/copies for each species or taxon. Red lines mark significant expansion of the DKK3 and PDCD5 gene copies. Comparison of protein sequence and 3D structure of DKK3 (C) and PDCD5 (D) using the visualization software PyMOL.

Article Snippet: Cells were treated with recombinant human DKK3 (hDKK3, 200 ng/ml), rat DKK3 (rDKK3, 200 ng/ml) or mouse DKK3 (mDKK3, 200 ng/ml) (Creative BioMart, Shirley, NY) for 24 hours.

Techniques: Comparison, Sequencing, Software

Journal: American Journal of Translational Research

Article Title: Use of tumor suppressor genes of naked mole rats for human cancer treatment

doi:

Figure Lengend Snippet: Anti-tumor effect and molecular mechanisms of DKK3 on breast cancer cells. A. Proliferation of human, rat, and mouse breast cancer cells with DKK3 treatment was measured using the MTT assay. B. Changes in β-catenin, p-β-catenin, p-AKT and p-mTOR in DKK3-treated breast cancer cells and their parental cells were studied using western blotting. C. Stemness regulatory transcription factors, SOX2, Nanog, C-myc and Oct4, in DKK3-treated breast cancer cells and their parental cells were studied using RNA sequencing.

Article Snippet: Cells were treated with recombinant human DKK3 (hDKK3, 200 ng/ml), rat DKK3 (rDKK3, 200 ng/ml) or mouse DKK3 (mDKK3, 200 ng/ml) (Creative BioMart, Shirley, NY) for 24 hours.

Techniques: MTT Assay, Western Blot, RNA Sequencing

Journal: American Journal of Translational Research

Article Title: Use of tumor suppressor genes of naked mole rats for human cancer treatment

doi:

Figure Lengend Snippet: Anti-tumor effect and molecular mechanisms of PDCD5 on breast cancer cells. A. Proliferation of human, rat, and mouse breast cancer cells with DKK3 treatment was measured using the MTT assay. B. Changes of β-catenin, p-β-catenin, p-AKT, and p-mTOR in PDCD5-treated breast cancer cells and their parental cells were studied using western blotting. C. Stemness regulatory transcription factors, SOX2, Nanog, C-myc and Oct4, in PDCD5-treated breast cancer cells and their parental cells were studied using RNA sequencing.

Article Snippet: Cells were treated with recombinant human DKK3 (hDKK3, 200 ng/ml), rat DKK3 (rDKK3, 200 ng/ml) or mouse DKK3 (mDKK3, 200 ng/ml) (Creative BioMart, Shirley, NY) for 24 hours.

Techniques: MTT Assay, Western Blot, RNA Sequencing