目的 观察冬虫夏草对高糖诱导的大鼠肾小球系膜细胞色素上皮衍生因子(PEDF)和血管内皮生长因子(VEGF)表达的影响,探讨其肾脏保护的作用机制.方法 35只8周龄清洁级雄性SD大鼠,体重260～ 280 g,根据体重按随机数字表法分为正常血清组(n=18)、低剂量虫草提取液喂养组(5 g· kg-1· d-1,n=8)和高剂量虫草提取液喂养组(10 g·kg-1·d-1,n=9),采用生理盐水及不同剂量虫草提取液灌胃8d后,分离静脉血提取对照血清及不同浓度的冬虫夏草含中药血清.体外培养HBZY-1大鼠肾小球系膜细胞,之后分别加入正常血糖正常血清[正常血糖组(NC组):5.6 mmol/L葡萄糖+10％胎牛血清(FBS)+1％正常大鼠血清]、高血糖正常血清[高血糖组(HG组):30.0 mmol/L葡萄糖+10％ FBS+1％正常大鼠血清]、低剂量虫草含药血清[低药组(LD):30.0 mmol/L葡萄糖+10％ FBS+1％低剂量虫草含药血清]和高剂量虫草含药血清[高药组(HD):30.0 mmol/L葡萄糖+10％ FBS+ 1％高剂量虫草含药血清]进行培养,采用逆转录-聚合酶链反应(RT-PCR)测定各组细胞PEDF及VEGF mRNA的表达,酶联免疫吸附试验(ELISA)测定各组细胞上清液中PEDF及VEGF的水平.组间资料比较采用单因素方差分析,样本间资料比较采用t检验.结果 与NC组相比,HG组PEDF mRNA表达明显下降(24 h:0.375±0.011比0.507±0.008,t=16.828,P＜0.01;72 h:0.376±0.014比0.515±0.010,t=14.034,P＜0.01),VEGF mRNA表达明显上升(24 h:1.005±0.011比0.864±0.012,t=14.963,P＜0.01;72 h:1.168±0.012比0.873±0.011,t=31.417,P＜0.01);LD组和HD组PEDF mRNA表达较HG组显著上调(24 h:0.505±0.018、0.639±0.012比0.375±0.011,F=354.719,P＜ 0.01;72 h:0.612±0.023、0.700±0.019比0.376±0.014,F=97.82,P＜0.01),VEGF mRNA表达明显下降(24 h:0.838±0.014、0.767±0.017比1.005±0.011,F=79.55,P＜ 0.01;72 h:0.923±0.016、0.784±0.012比1.168±0.012,F=244.28,P＜0.01),且HD组疗效更明显.经对照血清及CS血清培养24、72 h后,与NC组相比,HG组PDEF水平明显降低[24 h:(267±8)比(303±10) ng/L,t=6.493,P＜0.01;72 h:(265±27)比(338 ±42) ng/L,=3.259,P ＜0.01,VEGF明显升高[24 h:(128 ±3)比(113±8)ng/L,t=3.737,P ＜0.01;72 h:(144 ±7)比(125±7)ng/L,t =4.215,P＜0.01];与HG组相比,LD组和HD组PEDF明显升高[24 h:(297±12)、(296±26)比(267±8) ng/L,F=5.427,P＜0.01;72 h:(323 ±20)、(332±33)比(265±27) ng/L,F=5.690,P＜0.01],VEGF明显降低[24 h:(106 ±6)、(109±11)比(128±3) ng/L,F=5.738,P ＜0.01;72 h:(124±9)、(125±7)比(144 ±7) ng/L,F =7.878,P ＜0.01].结论 冬虫夏草可能通过降低肾小球系膜细胞VEGF的表达,升高PEDF的表达,对肾脏起到保护作用.
目的 觀察鼕蟲夏草對高糖誘導的大鼠腎小毬繫膜細胞色素上皮衍生因子(PEDF)和血管內皮生長因子(VEGF)錶達的影響,探討其腎髒保護的作用機製.方法 35隻8週齡清潔級雄性SD大鼠,體重260～ 280 g,根據體重按隨機數字錶法分為正常血清組(n=18)、低劑量蟲草提取液餵養組(5 g· kg-1· d-1,n=8)和高劑量蟲草提取液餵養組(10 g·kg-1·d-1,n=9),採用生理鹽水及不同劑量蟲草提取液灌胃8d後,分離靜脈血提取對照血清及不同濃度的鼕蟲夏草含中藥血清.體外培養HBZY-1大鼠腎小毬繫膜細胞,之後分彆加入正常血糖正常血清[正常血糖組(NC組):5.6 mmol/L葡萄糖+10％胎牛血清(FBS)+1％正常大鼠血清]、高血糖正常血清[高血糖組(HG組):30.0 mmol/L葡萄糖+10％ FBS+1％正常大鼠血清]、低劑量蟲草含藥血清[低藥組(LD):30.0 mmol/L葡萄糖+10％ FBS+1％低劑量蟲草含藥血清]和高劑量蟲草含藥血清[高藥組(HD):30.0 mmol/L葡萄糖+10％ FBS+ 1％高劑量蟲草含藥血清]進行培養,採用逆轉錄-聚閤酶鏈反應(RT-PCR)測定各組細胞PEDF及VEGF mRNA的錶達,酶聯免疫吸附試驗(ELISA)測定各組細胞上清液中PEDF及VEGF的水平.組間資料比較採用單因素方差分析,樣本間資料比較採用t檢驗.結果 與NC組相比,HG組PEDF mRNA錶達明顯下降(24 h:0.375±0.011比0.507±0.008,t=16.828,P＜0.01;72 h:0.376±0.014比0.515±0.010,t=14.034,P＜0.01),VEGF mRNA錶達明顯上升(24 h:1.005±0.011比0.864±0.012,t=14.963,P＜0.01;72 h:1.168±0.012比0.873±0.011,t=31.417,P＜0.01);LD組和HD組PEDF mRNA錶達較HG組顯著上調(24 h:0.505±0.018、0.639±0.012比0.375±0.011,F=354.719,P＜ 0.01;72 h:0.612±0.023、0.700±0.019比0.376±0.014,F=97.82,P＜0.01),VEGF mRNA錶達明顯下降(24 h:0.838±0.014、0.767±0.017比1.005±0.011,F=79.55,P＜ 0.01;72 h:0.923±0.016、0.784±0.012比1.168±0.012,F=244.28,P＜0.01),且HD組療效更明顯.經對照血清及CS血清培養24、72 h後,與NC組相比,HG組PDEF水平明顯降低[24 h:(267±8)比(303±10) ng/L,t=6.493,P＜0.01;72 h:(265±27)比(338 ±42) ng/L,=3.259,P ＜0.01,VEGF明顯升高[24 h:(128 ±3)比(113±8)ng/L,t=3.737,P ＜0.01;72 h:(144 ±7)比(125±7)ng/L,t =4.215,P＜0.01];與HG組相比,LD組和HD組PEDF明顯升高[24 h:(297±12)、(296±26)比(267±8) ng/L,F=5.427,P＜0.01;72 h:(323 ±20)、(332±33)比(265±27) ng/L,F=5.690,P＜0.01],VEGF明顯降低[24 h:(106 ±6)、(109±11)比(128±3) ng/L,F=5.738,P ＜0.01;72 h:(124±9)、(125±7)比(144 ±7) ng/L,F =7.878,P ＜0.01].結論 鼕蟲夏草可能通過降低腎小毬繫膜細胞VEGF的錶達,升高PEDF的錶達,對腎髒起到保護作用.
목적 관찰동충하초대고당유도적대서신소구계막세포색소상피연생인자(PEDF)화혈관내피생장인자(VEGF)표체적영향,탐토기신장보호적작용궤제.방법 35지8주령청길급웅성SD대서,체중260～ 280 g,근거체중안수궤수자표법분위정상혈청조(n=18)、저제량충초제취액위양조(5 g· kg-1· d-1,n=8)화고제량충초제취액위양조(10 g·kg-1·d-1,n=9),채용생리염수급불동제량충초제취액관위8d후,분리정맥혈제취대조혈청급불동농도적동충하초함중약혈청.체외배양HBZY-1대서신소구계막세포,지후분별가입정상혈당정상혈청[정상혈당조(NC조):5.6 mmol/L포도당+10％태우혈청(FBS)+1％정상대서혈청]、고혈당정상혈청[고혈당조(HG조):30.0 mmol/L포도당+10％ FBS+1％정상대서혈청]、저제량충초함약혈청[저약조(LD):30.0 mmol/L포도당+10％ FBS+1％저제량충초함약혈청]화고제량충초함약혈청[고약조(HD):30.0 mmol/L포도당+10％ FBS+ 1％고제량충초함약혈청]진행배양,채용역전록-취합매련반응(RT-PCR)측정각조세포PEDF급VEGF mRNA적표체,매련면역흡부시험(ELISA)측정각조세포상청액중PEDF급VEGF적수평.조간자료비교채용단인소방차분석,양본간자료비교채용t검험.결과 여NC조상비,HG조PEDF mRNA표체명현하강(24 h:0.375±0.011비0.507±0.008,t=16.828,P＜0.01;72 h:0.376±0.014비0.515±0.010,t=14.034,P＜0.01),VEGF mRNA표체명현상승(24 h:1.005±0.011비0.864±0.012,t=14.963,P＜0.01;72 h:1.168±0.012비0.873±0.011,t=31.417,P＜0.01);LD조화HD조PEDF mRNA표체교HG조현저상조(24 h:0.505±0.018、0.639±0.012비0.375±0.011,F=354.719,P＜ 0.01;72 h:0.612±0.023、0.700±0.019비0.376±0.014,F=97.82,P＜0.01),VEGF mRNA표체명현하강(24 h:0.838±0.014、0.767±0.017비1.005±0.011,F=79.55,P＜ 0.01;72 h:0.923±0.016、0.784±0.012비1.168±0.012,F=244.28,P＜0.01),차HD조료효경명현.경대조혈청급CS혈청배양24、72 h후,여NC조상비,HG조PDEF수평명현강저[24 h:(267±8)비(303±10) ng/L,t=6.493,P＜0.01;72 h:(265±27)비(338 ±42) ng/L,=3.259,P ＜0.01,VEGF명현승고[24 h:(128 ±3)비(113±8)ng/L,t=3.737,P ＜0.01;72 h:(144 ±7)비(125±7)ng/L,t =4.215,P＜0.01];여HG조상비,LD조화HD조PEDF명현승고[24 h:(297±12)、(296±26)비(267±8) ng/L,F=5.427,P＜0.01;72 h:(323 ±20)、(332±33)비(265±27) ng/L,F=5.690,P＜0.01],VEGF명현강저[24 h:(106 ±6)、(109±11)비(128±3) ng/L,F=5.738,P ＜0.01;72 h:(124±9)、(125±7)비(144 ±7) ng/L,F =7.878,P ＜0.01].결론 동충하초가능통과강저신소구계막세포VEGF적표체,승고PEDF적표체,대신장기도보호작용.
Objective To investigate the effects of Cordyceps sinensis (CS) on the expression of pigment epithelium - derived factor (PEDF) and vascular endothelial growth factor (VEGF) in high glucose induced rat mesangial cells (MCs).Methods 35 SD rats were randomly divided into three groups:normal serum group (Saline,n =18),low dose CS feeding group (5 g · kg-1 · d-1,n =8),and high dose CS feeding group (10 g · kg-1 · d-1,n =9).The three groups were given saline and different dosage of CS by gavage for 8 days,then venous blood samples were taken to prepare experimental serum.Rat mesangial cells (MCs,HBZY- 1 line ) were divided into four groups:the normal glucose control group (NC group,5.6 mmol/L Glucose + 10％ Fetal bovine serum(FBS) + 1％ normal serum),the high glucose group ( HG group,30.0 mmol/L Glucose + 10％ FBS + 1％ normal serum),the Low dose - CS group (LD Group,30.0 mmol/L Glucose + 10％ FBS + 1％ low dose CS serum),and the High dose -CS group (HD Group,30.0 mmol/L Glucose + 10％ FBS + 1％high dose CS serum).The VEGF and PEDF mRNA expression in the MCs were determined by reverse transcription - polymerase chain reaction ( RT - PCR),the level of PEDF and VEGF in MCs supernatants were examined by ELISA.Results Compared with the NC group,the mRNA expression of PEDF was significautly lower (24 h:0.375 ± 0.011 vs 0.507 ± 0.008,t =16.828,P ＜ 0.01 ;72 h:0.376 ± 0.014 vs 0.515 ± 0.010,t =14.034,P ＜ 0.01 ),whereas the VEGF mRNA expression was significantly higher in the HG group ( 24 h:1.005 ± 0.011 vs 0.864 ± 0.012,t =14.963,P ＜ 0.01 ;72 h:1.168 ± 0.012 vs 0.873 ± 0.011,t =31.417,P ＜ 0.01 ).After treated with CS,the PEDF mRNA expression was significantly increased (24 b:0.505 ± 0.018,0.639 ± 0.012 vs 0.375 ± 0.011,F =354.719,P ＜ 0.01 ; 72 h:0.612 ± 0.023,0.700 ± 0.019 vs 0.376 ± 0.014,F =97.82,P ＜ 0.01 ),and the VEGF mRNA expression was significantly decreased in the LD and the HD groups compared with those in the HG group (24 h:0.838 ±0.014,0.767 ±0.017 vs 1.005 ±0.011,F =79.55,P ＜0.01;72 h:0.923 ±0.016,0.784 ± 0.012 vs 1.168 ±0.012,F =244.28,P ＜0.01 ).After treated with experimental serum for 24 and 72 hours,the level of PEDF was significantly lower( 24 h:( 267 ± 8 ) vs ( 303 ± 10) ng/L,t =6.493,P＜0.01;72 h:(265±27) vs (338 ±42) ng/L,t=3.259,P＜0.01),whereas the level of VEGF was significantly higher in the HG group than those of the NC group ( 24 h:( 128 ± 3 ) vs ( 113 ± 8 ) ng/L,t =3.737,P ＜0.01 ;72 h:( 144 ±7) vs ( 125 ±7) ng/L,t =4.215,P ＜0.01 ).Compared with the HG group,the level of PEDF was significantly higher(24 h:(297 ± 12),(296 ± 26) vs (267 ± 8 ) ng/L,F =5.427,P＜0.01;72h:(323±20),(332±33) vs (265 ±27) ng/L,F=5.690,P＜0.01),and the level of VEGF was significantly lower in the LD and the HD groups ( 24 h:( 106 ± 6 ),( 109 ± 11 ) vs ( 128 ± 3 ) ng/L,F=5.738,P＜0.01;72 h:(124±9),(125 ±7) vs (144±7) ng/L,F=7.878,P＜0.01),no difference was observed between the LD group and the HD group ( P ＞ 0.05).Conclusion Renoprotection of CS on MCs may be mediated through reducing the expression of VEGF and increasing the expression of PEDF.
