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Objective To observe the effects of orexin-1 receptor inhibitor on proliferation and estrogenic differentiation of bone marrow mesenchymal stem cells ( BMSCs) . Methods BMSCs were isolated and cultured from Sprague-Dawley rats, and orexin-1 receptor inhibitor was prepared to different concentration of solutions:10-2 , 10-3 , 10-4 , 10-5 and 10-6 mol·L-1 . Then, the cells at passage 3 were cultured in orexin-1 receptor inhibitor solutions. The cell growth, alkaline phosphatase activity and osteocalcin secretion were measured by MTT, PNPP and ELISA, respectively. The mRNA expression levels of Runx2 and COL1A1 were detected by real-time quantitative PCR. Results The best concentration of orexin-1 receptor inhibitor for the proliferation of BMSCs was 1 × 10-4 mol·L-1 . Orexin-1 receptor inhibitor promoted the proliferation at concentration of 10-6 , 10-5 , 10-4 mol·L-1 in 3, 5 and 7 days. Orexin-1 receptor inhibitor at concentration of 10-6 , 10-5 and 10-4 mol·L-1 for 5 days significantly stimulated ALP activity. Orexin-1 receptor inhibitor significantly up-regulated Runx2 and COL1A1 mRNA expression at concentration of 10-6, 10-5, 10-4 mol·L-1 for 5 days (P<0.05). Conclusion Orexin-1 receptor inhibitor can promote BMSCs proliferation and stimulated ALP activity and osteocalcin secretion.
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Objective To observe the effects of orexin-1 receptor inhibitor on proliferation and estrogenic differentiation of bone marrow mesenchymal stem cells ( BMSCs) . Methods BMSCs were isolated and cultured from Sprague-Dawley rats, and orexin-1 receptor inhibitor was prepared to different concentration of solutions:10-2 , 10-3 , 10-4 , 10-5 and 10-6 mol·L-1 . Then, the cells at passage 3 were cultured in orexin-1 receptor inhibitor solutions. The cell growth, alkaline phosphatase activity and osteocalcin secretion were measured by MTT, PNPP and ELISA, respectively. The mRNA expression levels of Runx2 and COL1A1 were detected by real-time quantitative PCR. Results The best concentration of orexin-1 receptor inhibitor for the proliferation of BMSCs was 1 × 10-4 mol·L-1 . Orexin-1 receptor inhibitor promoted the proliferation at concentration of 10-6 , 10-5 , 10-4 mol·L-1 in 3, 5 and 7 days. Orexin-1 receptor inhibitor at concentration of 10-6 , 10-5 and 10-4 mol·L-1 for 5 days significantly stimulated ALP activity. Orexin-1 receptor inhibitor significantly up-regulated Runx2 and COL1A1 mRNA expression at concentration of 10-6, 10-5, 10-4 mol·L-1 for 5 days (P<0.05). Conclusion Orexin-1 receptor inhibitor can promote BMSCs proliferation and stimulated ALP activity and osteocalcin secretion.
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Objective To investigate clinical efficacy of tissue-engineered artificial nerve grafts constructed by acellular nerve grafts combined with adult rat Schwann cell (SC)in the treatment of peripheral nerve injury.Methods SCs were isolated and cultured from the distal nerves of adult Sprague Dawley (SD) rats with 1-week Wallerian degeneration and then combined with acellular nerve grafts to construct tissue-engineered artificial nerve.All rats were divided into acellular nerve graft containing SCs (SC group)and nerve graft containing no cells groups (control group),five animals in each group.At 2-,4-and 8-week after surgery,sciatic function index (SFI)of the affected side was compared between two groups.At postoperative 8 weeks,nerve conduction of sciatic nerve of the injured side,recovery rate of triceps surae wet weight and other relevant parameters were equally compared between two groups.Results In the SC group,SFI of the affected side at 2-,4-and 8-week after surgery,nerve conduction of sciatic nerve at the injured side and recovery rate of triceps surae wet weight at postoperative 8 weeks were significantly better compared with those in the control group (all in P <0.05).Conclusions Combined use of adult rat SCs and acellular nerve grafts effectively repairs peripheral nerve defects and accelerates functional recovery of injured nerves.
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Objective To discuss the effect of bone marrow mesenchymal stem cell (BMSC)as the seed cell transplantation of tissue-engineered artificial nerve in the treatment of peripheral nerve injury. Methods BMSC was obtained from the bone marrow of adult rat through isolation and culture and combined with acellular nerve scaffold to construct ‘tissue-engineered artificial nerve’.After transplantation,rats were divided into two groups,the BMSC +acellular nerve conduit group(BMSC treatment group)and the empty cell conduit group(negative control group)with 5 rats in each group.Sciatic functional index (SFI)of the affected side of rats was compared between two groups at 2 weeks,4 weeks and 8 weeks after the surgery.Moreover,the sciatic conduction,recovery rate of tricipital muscle wet weight and other repair effects of the affected side were compared between two groups at 8 weeks after the surgery.Results The indicators of BMSC treatment group, including SFI assessed at 2 weeks,4 weeks and 8 weeks after the surgery as well as the sciatic conduction and recovery rate of tricipital muscle wet weight assessed at 8 weeks after the surgery,were better than those of the negative control group(all in P <0.05).Conclusions BMSC combined with tissue-engineered artificial nerve of acellular nerve scaffold can effectively promote nerve regeneration and function recovery.
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AIM:To explore the effects of panax notoginsenosides(PNS)on bone histomorphometry in rats with hyperlipidemia-induced osteoporosis.METHODS:Thirty 3-month-old Sprague-Dawley rats were randomly divided into three groups with 10 per group as follows.Control group was intragastrically given distilled water at the dose of 5 mL?kg-1?d-1 for 20 weeks;Model group was intragastrically given lipid emulsion at the dose of 5 mL?kg-1?d-1 for 20 weeks;PNS preventing group was intragastrically given lipid emulsion at the dose of 5 mL?kg-1?d-1 in the morning and PNS at the dose of 5 mL?kg-1?d-1 at afternoon for 20 weeks.At the end of the experiment,the right longitudinal proximal tibial metaphyseal parameters of rats were performed undecalcifiedly and used for bone histomorphometry analysis.And the blood serum contents of FFA,TG,TC,HDL-C and LDL-C were measured by biochemical analysis.RESULTS:Compared with model group,the blood serum contents of FFA,TC and LDL-C were decreased and HDL-C was increased significantly(P