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Therapeutic Effect of Rosiglitazone Combined with Adipose-Derived Stem Cell Transplantation in Rats with Type 2 Diabetes Mellitu

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Therapeutic Effect of Rosiglitazone Combined with Adipose-Derived Stem Cell Transplantation in Rats with Type 2 Diabetes Mellitu ęඈඋ1࣏ۘऺ ৰ 21 Ż ৰ 33 ƛ 20171128 ϦČ Chinese Journal of Tissue Engineering Research November 28, 2017 Vol.21, No.33 www.CRTER.org ऺƣጛۘ 2Ƨசɳ҉۔ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћ̓ ăএ) ϴ ዐ(ǧSďS˂ˡŋʒ½ˍãKǧSď 300350) .Ƨசɳ҉[J].ęඈඋ1࣏ۘऺ 2017 21(33):5325-5331 2 ۔šϬƨăএ) ϴዐ. ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћ̓ DOI:10.3969/j.issn.2095-4344.2017.33.013 ORCID: 0000-0001-9343-6732(ăএ)) তȷợⓉᪿ ÒŻűK˿K1973 ˝ÛK Ƨசɳ҉ 2 ۔ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћ̓ ǧSďNK˩ȂKƶƍ åKƶϙÎāˍǫɯΡ~ ̼ϟūʃ Ƿ ͏ћȒ 3 ɬ Ě ିǻ:R394.2 ाွᜄ ඗Ș۔ȭ+ඈ (1)ᢆȓȈඈ̓ B:ۅǒɀ¬ĭ Dzʃ᪊ (1)µƧඈ˸྇ǎɲ☝࿍ ச3ྴˏ௤3CཱྀǎĿᯬ ปʸ′⍤ █ɳ SD ̓ তේǻ:2095-4344 ˈ֔ˊͼȸϣ ₓǜĚ ༘Ȍ࿆཮¾ (2017)33-05325-07 ปʸ′⍤ඈ˸྇ඝ(2) ۔ (2)࿆཮¾ඊྡྷ Oǎƌ ඊྡྷࢿΉdz ɘ4ěY2017-06-03 tปʸ′⍤ Ϗ ƅά͏ћ̓ 2 Ƨசɳ Ƨசɳ 2 ۔ ཮¾ඊྡྷඈɲ☝࿍ (3)ྴှඈඋƮɍǜĚǎ࿆(3) ҉µƧ ͼȸ࿆཮¾ඊྡྷ ྴˏඊྡྷη̑Ρ ҉ õ:̺ (4)ǒɀඈ˸྇ඝtปʸ (4) ྴ ှ ඈ උ CXCL1 3 ҋǑà4 ′⍤ՈȐrɲ☝࿍ͼȸ CXCL2 ǎ͢ǛĿ CXCR2 ࿆཮¾ඊྡྷ4 ᖏՁᜬẂˈ¿4 L₎5 ปʸ′⍤«P࢑ ÖZϬՈ̕Σx′ିሳĭ dzƅά┑ĺ 2 Ƨசɳ҉˯້Ոᜄசˈ¿ ෗ᢧ̺ҋǑ à IJ⑃ƛūϬKĆȰႸཀྵྚʐˈྃ਍]ᄳǑà4 ཮¾ඊྡྷࢿΉ͏ћசɳ҉dz&ƶĿȴƇǷՈྴˏඈඋඊྡྷ ẟ໐ŻÏĿͱ>Ǜ4۸ūՈྴˏඈඋ ම࿆ ťǷՈᜄசˈ¿ ęû̺ҋǑà ෗ᢧசɳ҉ՈÂǕҋ਍ IJʌசɳʟȭ࿆཮¾ඊྡྷͣƅǃĐňϬ ƽˑ ࿆཮¾ඊྡྷࢿΉ͏ћՈάȘ4 ʼᡅ ëۘऺº߾ ʌசɳʟȭ࿆཮¾ඊྡྷƌϏͣƅºՈõ:ňϬ ẝPňϬₑƽˑ࿆཮¾ඊྡྷń 2 Ƨசྐ ɳ҉͏ћ5☦ՈάȘ4 Ƨசɳ҉ՈάȘ4 2 ۔Ոᢆȓปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћֲ̓ ỞẋွဘͼȸɃFₓ⏂࿶◍௤༘۔Ǯ ╓ƶǚ 20 Ǯň&Ƿȭ+ඈ mŅՈ 88 Ǯ̓ 108 ۔̓ 5ͩǚ SD ˈ֔ˊ 4 ඈ͏ћµƧඈ˸྇ǎɲ☝࿍ͼȸϣ Ȍʌசʌ࿆Ģņএ 2 Ƨசɳ҉µƧ ỤµȒ 20 d ╓ƶ ปʸ′⍤ඈ˸྇ඝtปʸ′⍤ ࿆཮¾ඊྡྷඈɲ☝࿍ͼȸ࿆཮¾ඊྡྷ ǒɀඈ˸྇ඝtปʸ′⍤ՈȐ ाွᜄச3ྴˏ௤3Cཱྀ۔rɲ☝࿍ͼȸ࿆཮¾ඊྡྷ4ţ 1 Ƶ/d Ảන͏ћ 7 d4͏ћȒ 3 ɬ ᢆȓȈඈ̓ ǎĿᯬₓǜĚ ࿆཮¾ඊྡྷ OǎƌϏ ྴှඈඋƮɍǜĚǎྴˏඊྡྷη̑Ρ Ñǎྴှඈඋ CXCL13 CXCL2 ǎ͢ǛĿ CXCR2 ᖏՁᜬẂˈ¿4 ඗Ș^඗᩾ ^Ƿȭ+ඈɨṇ µƧඈाွᜄசŋʌ(P < 0.05) ྴˏ௤3CཱྀǎĿᯬₓţ[┑(P < 0.05) ྴˏඊྡྷηϔₓʺ¤(P < 0.05) ྴှඈඋ CXCL13CXCL2 ǎ͢ǛĿ CXCR2 ᖏՁᜬẂŋʌ(P < 0.05) ྴ ˏඈඋƮɍ௎] ±ᢅɅₓྴˏඊྡྷþ ^µƧඈɨṇ ปʸ′⍤ඈ3࿆཮¾ඊྡྷඈ3ǒɀඈाွᜄச ┑ĺ(P < 0.05) ྴˏ௤3CཱྀǎĿᯬₓţŋʌ(P < 0.05) ྴˏඊྡྷηϔₓλɅ(P < 0.05) ྴှඈඋ CXCL13 CXCL2 ǎ͢ǛĿ CXCR2 ᖏՁᜬẂ┑ĺ(P < 0.05) ྴˏඈඋƮɍɌɴ]Ȑ࣏òΝΈ ͢ÑǒɀඈȈūʃǎ ྴˏඈඋΝΈ࣏òżº ࿆཮¾ඊྡྷඈ3ǒɀඈdzᢅࢿΉՈ࿆཮¾ඊྡྷ Ñǒɀඈɹ̶඗Șᜬ Ƨசɳ҉ õ:̺ҋǑà4 2 ۔ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉdzƅά͏ћ̓ ͟⏲᪑ ҋᱏĚüƄ̺۔ඊྡྷ࿆཮¾ඊྡྷปʸ′⍤ࢿΉசɳ҉̓¾ 'L᪑ ¾ඊྡྷசɳ҉ඊྡྷࢿΉඈඋ1࣏ Therapeutic effect of rosiglitazone combined with adipose-derived stem cell transplantation in rats with type 2 diabetes mellitus Kang Li-li, Attending Kang Li-li, Tian Meng (Department of Internal Medicine, Tianjin Jinnan District Xianshuigu Hospital, Tianjin 300350, physician, Department of China) Internal Medicine, Tianjin Jinnan District Xianshuigu Hospital, Tianjin 300350, Abstract China BACKGROUND: Studies have shown that high glucose environment has a significant inhibitory effect on the ISSN 2095-4344 CN 21-1581/R CODEN: ZLKHAH 5 3 2 5 Ƨசɳ҉ www.CRTER.org 2 ۔ăএ) ਍. ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћ̓ survival of adipose-derived stem cells, which significantly affects the therapeutic efficacy of adipose-derived stem cells on type 2 diabetes mellitus. OBJECTIVE: To investigate the therapeutic effect of rosiglitazone combined with adipose-derived stem cell transplantation in rats with type 2 diabetes mellitus. METHODS: A total of 108 male Sprague-Dawley rats were chosen, 20 of which were randomly taken as normal controls and the rest 88 rats were used to make type 2 diabetes mellitus models through intraperitoneal injection of low-dose streptozotocin combined with high-sugar/high-fat diet. At 20 days after modeling, the model rats were randomized into model, rosiglitazone, cell transplantation and combined groups, followed by administration of normal saline (by gavage and via tail vein), rosiglitazone (by gavage), adipose-derived stem cells (via tail vein), and rosiglitazone (by gavage)+adipose-derived stem cells (via tail vein), once a day, for consecutive 7 days. Three weeks after treatment, fasting blood glucose level, insulin level, C-peptide level, body mass changes were detected; distribution and survival of adipose-derived stem cells were observed; the changes of pancreas histomorphology was observed, the apoptosis in islet cells was detected, and the CXCL1, CXCL2 and CXCR2 protein expression levels in the pancreas tissue were determined. RESULTS AND CONCLUSION: Compared with the normal control group, the fasting blood glucose level was increased (P < 0.05), while the insulin level, C-peptide level and body mass were significantly decreased in the model group (P < 0.05); the number of apoptotic islet cells was increased (P < 0.05); the expression levels of CXCL1, CXCL2 and CXCR2 in the pancreas tissue were elevated (P < 0.05); and the pancreas tissue morphologies were in disorder, with few islet cell masses. Compared with the model group, the fasting blood glucose level was decreased (P < 0.05), while the insulin level, C-peptide level and body mass were significantly increased in the rosiglitazone, cell transplantation and combined groups (P < 0.05); the number of apoptotic islet cells was decreased (P < 0.05); the expression levels of CXCL1, CXCL2 and CXCR2 in the pancreas tissue were declined (P < 0.05); and the pancreas tissue morphologies were improved to different extents, and the combined group showed the best improvement. There were many transplanted adipose-derived stem cells in the cell transplantation and combined groups, especially in the latter one. Overall findings indicate that rosiglitazone combined with adipose-derived stem cell transplantation is an effective treatment for type 2 diabetes mellitus in rats, which is better to inhibit inflammations. Subject headings: Stem Cells; Diabetes Mellitus; Cell Transplantation; Tissue Engineering Cite this article: Kang LL, Tian M. Therapeutic effect of rosiglitazone combined with adipose-derived stem cell transplantation in rats with type 2 diabetes mellitus. 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Box 10002, Shenyang 110180 www.CRTER.org Ƨசɳ҉ www.CRTER.org 2 ۔ăএ) ਍. ปʸ′⍤༘Ȍ࿆཮¾ඊྡྷࢿΉ͏ћ̓ 60 ?ʞÕˈ ੵ 30 sͱẟᜐȷợ̩ᆓ4ᙑĚ7ȒȺ࿆཮ ྡྷʃᩴɋ[22-24]4 ȏ̠̓⁍ٯඊྡྷẉ҂Ḱ͑EP੥¤͑DMEMȕͧȖ ₑ̩ɃȏȽ· ᆓƤஂ.ö൦ɏᄶᢆȓྴှඈඋƮɍǜĚ¾ ϔ10%϶ Ėǎྴှඈඋ ÂǚϦྴှඈඋ ϬĿࢳ۔ O ╓ȒࡿȏŏZ±^4Ⱥ̩ᆓȒՈඊྡྷϬ ඊྡྷū͢ţĄ DMEMǀ͔ȕͧȖ᫇Ϙ&1×109 L1Ոඊྡྷϧò4ẟᜐȕͧ3 :̣ྴှඈඋ ċūϬϵĺϧò4ʌϧòՈE⁋ň࿵ˈ ÏȒ ྴᖏՁ›Ţ^4Ě2.0 3.0 minȒඌǶ4ĚࡿȏŏZ F Ổ¼࿵ƿඈඋųՈˈ 4͹Ⱥඈඋųาz^ŢzE ᗥՈồFx϶ᆳồ Ñx϶ᆳŻdžϦඈڷញႷ᪙੥ ⁋njŢz ₑ˸ඊྡྷ4᫇ϘඊྡྷϧòႷ1×109 L1± û¦ උųՈE⁋ ╓Ȓẟᜐ$ᗥĉǧ4ȺĉǧΙՈᗥųĖǎ۔mL/੥4 Ȉ¤͑5 µL඗Ȍƅ FITCzʃᩴՈɃ 0.1 CD343CD293CD443CD4453 CD903CD105řû 4 ? z ċƶZ t5 8 µmƞՈᑈċ4ȺᑈċΗ¿ ͹᯸4 ὃ̵ƩĚ 30 minȒ ÑPBSΫP1Ƶ 1 000 r/minࡿȏ5 min ṁɿċZ 45 ?ʞÕ΄¾4࿵ᗥȒ ẟᜐᆓƤஂö൦ɏ Ϭ500 µL PBSₑ˸ඊྡྷ ϕśඊྡྷÖ̼ϟỤᜄඊྡྷᜬ☦ʃᩴ ᄶ4ɏᄶȒՈ ċඓ൳E⁋࿵ˈ ͹ඓx϶ᆳū ċồ4 ċƠZ¤ţ̓ʍྺ ֚Z֚ɿċȵĖ4Ǒʍ ĭՈᜬẂ4 Ⱥ>ồՈ ှඈඋՈƮɍǜྴ۔ℸǎǀtȒ ࡿȏƿZ±ΫP Ⱥ࿆཮¾ඊྡྷϧò᫇ ྺ¾©Ȓ ራ̵ºǺ␠[ᢆȓȈඈ̓ Ϙ&8×108 L1 ╓Ȓȕඊྡྷ˸^¤͑5 µL CM-DiLʃᩴ Ě4 ǚྴှඈ۔30 minȒ ࡿȏŏZ TUNEL̼ͩϟྴˏඊྡྷη̑Ρ̠ȏ̓ ඊྡྷ╓ȒาzȕͧੵʃᩴƩĚ25 ċᢈ࿵ᗥ ᗥڷ ċ͝ᩥϔ5 ှඈඋྴ۔PBSₑ˸ ₑ̩ʃᩴ2Ƶ ºǺ␠[ᢆȓඊྡྷʃᩴՈ̑ උ ɣǮ̓ ± Ρ4␠[dzᢅ࿆཮¾ඊྡྷʃᩴºɴϦѐᄷՈì൦ᄶhPᄶ ႷˈƠ¤1Ơ̩Ȍ4Ě^(ᖏՁ›K^྇ᖏՁ›) 25 ?Ʃ ţĄ ̣Ϭ4 Ě20 min ΫPȒƠ¤TUNELʃᩴǑà£Ȍ^50 µL ń ƶ ī֖37 ?Ʃྲྀ1 h Ơ¤POD2ḰĚ^50 µL ī֖ 37╓۔ඈ¾8 º108ǮSD̓ சɳ҉Ụµǎ 1.4.2 ö൦̩ Ảන₋7ɬՈʌ ?Ʃྲྀ30 min PBSΞΫ3Ƶ DABºᄶᆓƤஂ۔ọǚ20Ǯň&Ƿȭ+ඈ Ņ88ǮSD̓ ᄶඊྡྷʴՈඊྡྷ&ηඊྡྷ ᩥϔٸசʌ࿆Ģ(ʌ࿆ʌசĦȯ10%Ꮫச310%Ǯ͍ǎ5%ྊĖ ɏ ºǺ␠[dzᢅ͑ ⁋)7Ȓ Ảන3 dႮွဘͼȸɃFₓ⏂࿶◍௤(45 mg/kg) ηඊྡྷϔₓÂᩥਜ਼ඊྡྷηɋ[25-26]4 ņএ2Ƨசɳ҉µƧ4ỤµȒ20 d ỞẋᜄசÖ̼ϟỤµ̓ Western blot̼ͩϟྴှඈඋCXCƧᱏĚüƄ‑Ŀ ۔Ոɲ☝࿍ᜄ^Ոᜄசˈ¿ Ⱥ̼ϟ඗ȘϧඌÑȭ+Ոᜄ CXCL13CXCL2ǎ͢ǛĿCXCR2ᖏՁᜬẂˈ¿ǚ̓۔ சȨ(16.7 mmol/L)ň&ņµt£Ոʃβ4 ྴှඈඋ ʵǒBCA᪙F֖ȴǚÂϟǎྴှඈඋᖏՁϧò &4ඈ ɣඈ19 ੄໐ᣄ7Ⱥඊྡྷឆᢧ7ÂȺ7ࣄ₎ႷּȐՈᖏՁϧò ƶ╓۔Ⱥżඌņµt£Ո76ǮSD̓ ǮµƧඈ˸྇ඝtϣˊ֔ˈ1 mL ɲ☝࿍ͼȸ1 mLϣˊ 100 ?2͌5 min ūᖏՁǜɳ4ɣඈǚᖏՁ40 µg 10% ปʸ′⍤ඈ˸྇ඝt4 mg/(kg࠽d)ปʸ′⍤(1 mL SDS-PAGEωྺϹ·(70 V 30 min100 V 90 min)Ḱˈ֔ ϣˊ֔ˈŢᢧ) ɲ☝࿍ͼȸ1 mLϣˊ֔ˈ࿆཮¾ඊྡྷඈ ၠ(200 mA 3 h)ႷPVDFၠ5%࿵࿆ğΒǘÕȵ⒱2 h(z ඝt1 mLϣˊ֔ˈ ɲ☝࿍ͼȸ1 mL࿆཮¾ඊྡྷ˸^ 4 ?ẋ̸)Pû1a500 ǘÕƩྲྀ2 h (z4 ?ẋ̸)TPBS྇˸ (ඊྡྷϧò1×107 L1) ǒɀඈ˸྇ඝt4 mg/(kg࠽d)ปʸ ΫP3Ƶ PBSΫP1Ƶ7ȒẟᜐᖏՁ̹ѯŴẽՈǑà Ȍ ϟǎȈඈྴှඈඋ ⍤(1 mLϣˊ֔ˈŢᢧ) ɲ☝࿍ͼȸ1 mL࿆཮¾ඊྡྷ˸ ı+ᩴơ7ȒẟᜐḳâՈĚʻ′ ^)¿ˈඊྡྷϧò1×107 L1)4ȈඈẢන¾87 d 1Ƶ/d4 CXCL13CXCL2ǎ͢ǛĿCXCR2ᖏՁᯬּȭᜬẂ)^ 1.5 ƶϙʬ+ŷ/ ͏ћȒ3ɬ ẟᜐÑ[̼ϟ4 β-actinՈ̜òࢳ ɨȨ) ň&CXCL13CXCL2ǎ͢ǛĿ ाွᜄச3ྴˏ௤3CཱྀǎĿᯬₓǜĚ̼ϟ CXCR2ᖏՁᜬẂՈₓ4۔̓ Ȍα͸ǫʭ ¤ƅϔǒᰈՈඈⒸɨṇ₋Ϭřü௤ 1.6 ۔ाွᜄசʐĿᯬₓǜĚ(ń̼ϟ7Q◄ᡅȭ̓۔Ȉඈ̓ _ Ȍ ǒɀ඗ȘÑx±sᜬ߾ Ո 5:(One-way ANOVA)࣏Û۔ẟᜐ12 hՈाွ̠ˊ)4ाွᜄசỞẋᜄசÖ̼ϟ̓ ဘ☝ ÂᢈǎP < 0.05&:Ŏƅºጛɳ͛54]۔z̓ ۔ɲ☝࿍ᜄՈᜄசȯₓ ╓Ȓ̠ȏ̓ ཱྀ ाွྴˏ௤ʐC۔࿍ġᜄ ₋ϬELISA᪙F֖ϟǎ̓ ˈ¿4 2඗ȘResults ራ̵ºǺ␠ᢆȓCM-Dil⓷ɳඊྡྷ OǎƌϏ₋▊̓ 2.1 ūǓȼˍFĝ́Ƨʬ+ ǒɀọǚĿ̲ȕͧႷP3Ï Ոϣ⑃v֟Ո࿆཮¾ඊྡྷ ΞĚ1¤߾ ৰ3Ï࿆཮¾ඊ ċÂẟᜐ4’ 6 ှඈඋΜΧྴ۔ှඈඋ :ň̓ྴ۔ x࿖Ȗ 2ᆳȖȶ˞ɏᄶ4¾©Ȓ ºǺ␠[ᢆȓr ╓ ྡྷȕͧYͅȒ ඊྡྷ¿⎾᱉ẋκá80%ÑZ ඊྡྷɌ ƶọǚɣŬΣΧ ċՈ3ᢊₒhȭ7ẟᜐı+ᩴơȳ ▊ጁʳϣ⑃ ʴ­º ඗Ȁ±ì ƮɍĨt൨මʳඊ ᪂ɣŬ+ċՈʇò&1 ɣŬ+ċọǚP̵òͥ ྡྷ ඊྡྷᜬ☦̵Ž Oṇ&ţĄ dzᢅºǕMźՈ -ϬACDsee5.0ḳâܲǎ͢ŬʃȨ4ńĚʻ╓ƶọǚ8 ඊྡྷ̷╊ ƾඊྡྷȖƨ4͍Čৰ3ÏՈ࿆཮¾ඊྡྷºɴ ᢊₒ ᩥϔɣᢊₒ7CM-DilʃᩴՈ⓷ɳඊྡྷϔǎ ϦѐᄷՈì൦ᄶhPᄶţĄ4࿆཮¾ඊྡྷՈᜬ☦ʃᩴĭ ᢊₒ࿆཮¾ඊྡྷʇϔ Âᩥਜ਼ྴှඈඋՈ࿆཮¾ඊ ᜬẂ ᢅĚ24 ISSN 2095-4344 CN 21-1581/R CODEN: ZLKHAH 5327 Ƨசɳ҉ www.CRTER.org 2 ۔ăএ) ਍.
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