npsþsha68  (MedChemExpress)

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 6 PLAU and TIMP1 are enriched in USC-EVs and contribute to the anti-aging effects of USC-EVs in senescent cells. (A) Expression ratios of the top five most abundant anti-aging proteins in USC-EVs (E) compared with USCs (C) assessed by proteomic analysis. n Z 3 per group. (B) Inhibitory efficiency of shRNAs targeting PLAU or TIMP1 in USCs by qRT-PCR. n Z 3 per group. (C) Down-regulation efficiency of PLAU and TIMP1 in USC-EVs detected by Western blotting. (D) SA-b-gal staining images of different cell types in hydrogen peroxide con- taining medium added with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs. Scale bar: 50 mm. (E) Quantification of the mean intensity for the SA-b-gal-stained cells. n Z 3 per group. (F) SA-b-gal staining images of CAD cells treated with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs in serum-free condition. Scale bar: 50 mm. (G) Quantification of the mean intensity for the SA-b-gal-stained cells and the length of neurites. n Z 3 per group. (H) Quantification of the percent of late apoptotic/dead cells (Q2) and early apoptotic cells (Q3) tested by flow cytometry with annexin V-FITC/PI staining. n Z 3 per group. Data are shown as mean SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Cytometry

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 7 Inhibition of MMPs, P16INK4a, and P21Cip1 by PLAU and TIMP1 contributes to the anti-senescent effects of USC-EVs. (A, B) Western blot analysis of the protein expression of MMP1, MMP12, P16INK4a, P21Cip1, P-PI3K, and PI3K in HL-1, C2C12, and BMSCs receiving different treatments under senescence induction by hydrogen peroxide. (C) SA-b-gal staining images in BMSCs receiving different treatments under senescence induction by hydrogen peroxide. Scale bar: 50 mm. (D)e(F) Quantification of the mean intensity for the SA-b-gal-stained cells in BMSCs (D), C2C12 (E), and HL-1 (F) receiving different treatments under senescence induction by hydrogen peroxide. n Z 3 per group. Data are shown as mean SD. *P < 0.05, ***P < 0.001.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: Inhibition, Western Blot, Expressing, Staining

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 8 Downregulation of PLAU and TIMP1 weakens the benefits of USC-EVs on behavioral responses and bone quality in natural aging mice. (A)e(C) MWM test of the spatial learning and memory abilities of the natural aging mice treated with vehicle, USCshCon-EVs, USCshPLAU- EVs, or USCshTIMP1-EVs. n Z 8e10 per group. (D)e(F) Muscle strength and motor function assessments by grip strength test (D), balance beam test (E), and rotarod test (F) in natural aging mice treated with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs. n Z 8e10 per group. (G, H) mCT-reconstructed images of the femurs (G) and quantification of the trabecular and cortical bone microstructural parameters (H) in natural aging mice treated with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs. Scale bar: 1 mm. n Z 8e10 per group. (I) Three- point bending test of the femur strength in natural aging mice treated with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs. n Z 8e10 per group. Data are shown as mean SD. For panel (D): *P < 0.05 vs. Control group and #P < 0.05 vs. USCshCon-EVs group. */#P < 0.05, **/##P < 0.01, ***/###P < 0.001. For other panels: *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: Control

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 9 PLAU and TIMP1 mediate the inhibitory effects USC-EVs on cellular senescence in different organs of natural aging mice. (A, B) SA-b-gal staining images of the hippocampus (A) and quantification of the mean staining intensity (B) in natural aging mice treated with vehicle, USCshCon-EVs, USCshPLAU-EVs, or USCshTIMP1-EVs. Scale bar: 500 mm. n Z 5 per group. (C, D) Immunostaining images of P16INK4a in SGZ region of the hippocampus (C) and quantification of the mean staining intensity (D) in natural aging mice from (A). Scale bar: 100 mm. n Z 5 per group. (E, F) Immunostaining images of P21Cip1 in femur, muscle, heart, and skin tissues (E) and quantification of the mean staining intensity (F) in natural aging mice with different treatments. Scale bar: 50 mm (for femurs) or 100 mm (for other tissues). n Z 5 per group. Data are shown as mean SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: Staining, Immunostaining

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 10 PLAU and TIMP1 contribute to the USC-EVs-induced suppression of various aging-related phenotypes in different organs of natural aging mice. (A, B) TUNEL staining images (A) and quantification of apoptotic cell number (B) in bone tissues of the natural aging mice with different treatments. Scale bar: 20 mm. n Z 5 per group. (C, D) H&E staining images of muscle tissues (C) and quantification of muscle fiber size (D) in natural aging mice with different treatments. Scale bar: 100 mm. n Z 5 per group. (E, F) Immunohistochemical staining for COL17A1 in skin tissues (E) and quantification of the mean staining intensity (F) in natural aging mice with different treatments. Scale bar: 50 mm. n Z 5 per group. (G, H) Masson’s trichrome staining of skin tissues (G) and quantification of the mean staining intensity as well as dermal thickness (H) in natural aging mice with different treatments. Scale bar: 200 mm. n Z 5 per group. (I) Evaluation of the serum concentrations of SASP factors in natural aging mice with different treatments by ELISA. n Z 5 per group. Data are shown as mean SD. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: TUNEL Assay, Staining, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay

Journal: Acta pharmaceutica Sinica. B

Article Title: Extracellular vesicles from human urine-derived stem cells delay aging through the transfer of PLAU and TIMP1.

doi: 10.1016/j.apsb.2023.12.009

Figure Lengend Snippet: Figure 11 Schematic diagram showing human USC-EVs rejuvenate multiple old organs in aged mice through the transfer of PLAU and TIMP1 proteins. USCs were isolated from human urine and USC-EVs were collected from the culture medium of USCs. The intravenous administration of USC-EVs to aged mice reduced senescent cell burden and improved the structure and function of various organs by transferring PLAU and TIMP1 proteins, thereby rejuvenating the old organ towards a more “youthful” state.

Article Snippet: To assess whether the inhibition of MMPs or PI3KeAkt signaling could affect the anti-senescent effects of USC-EVs, PLAU, and TIMP1, the cells were treated with hydrogen peroxide with or without PLAU, TIMP1, ilomastat (10 nmol/L; Selleckchem, Houston, USA), wortmannin (10 nmol/L; MedChemExpress, NJ, USA), USC-EVs, ilomastat þ USC-EVs, or wortmannin þ USC-EVs.

Techniques: Isolation, Transferring