<TR>
<TD class=line18 vAlign=top align=middle>
<><FONT size=5>镇银膏药理学报告之四:镇银膏对NK细胞活性的影响(山东医科大学)</FONT></P></TD></TR>
<TR>
<TD class=line18 vAlign=top>
< ><FONT size=5><FONT face="Times New Roman"> 1</FONT>.试验方法<FONT face="Times New Roman"> </FONT>取<FONT face="Times New Roman">BALB/C</FONT>小鼠<FONT face="Times New Roman">20</FONT>只,体重<FONT face="Times New Roman">18</FONT>~<st1:chmetcnv UnitName="g" SourceValue="22" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on"><FONT face="Times New Roman">22g</FONT></st1:chmetcnv>,雌雄各半,随机分为<FONT face="Times New Roman">2</FONT>组,每只鼠背部剃毛约<FONT face="Times New Roman"><st1:chmetcnv UnitName="cm" SourceValue="3" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on">3cm</st1:chmetcnv><SUP>2</SUP></FONT>,第<FONT face="Times New Roman">1</FONT>组为空白对照组,于背部剃毛处涂抹芝麻油<FONT face="Times New Roman">0</FONT>.<FONT face="Times New Roman">05ml</FONT>/<st1:chmetcnv UnitName="g" SourceValue="10" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on"><FONT face="Times New Roman">10g</FONT></st1:chmetcnv>,第<FONT face="Times New Roman">2</FONT>组为镇银膏组,于背部剃毛处涂抹镇银膏<FONT face="Times New Roman">0</FONT>.<FONT face="Times New Roman">05ml</FONT>/<st1:chmetcnv UnitName="g" SourceValue="10" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on"><FONT face="Times New Roman">10g</FONT></st1:chmetcnv>,隔日<FONT face="Times New Roman">1</FONT>次共<FONT face="Times New Roman">7</FONT>次。按常规方法取脾脏并制成单个核细胞悬液,以<FONT face="Times New Roman">1640</FONT>完全培养液(<FONT face="Times New Roman">CM</FONT>)调整细胞浓度为<FONT face="Times New Roman">l</FONT>×<FONT face="Times New Roman">10<SUP>8</SUP></FONT>/<FONT face="Times New Roman">ml</FONT>作为效应细胞。靶细胞为传代<FONT face="Times New Roman">72h</FONT>培养的<FONT face="Times New Roman">L<SUB>929</SUB></FONT>细胞,经洗涤,用<FONT face="Times New Roman">CM</FONT>调整靶细胞浓度为<FONT face="Times New Roman">l</FONT>×<FONT face="Times New Roman">10<SUP>6</SUP></FONT>/<FONT face="Times New Roman">ml</FONT>细胞浓度。取上述靶细胞悬液,加入<FONT face="Times New Roman">96</FONT>孔平底培养板,每孔<FONT face="Times New Roman">0.1ml</FONT>;放于<FONT face="Times New Roman">37<SUP>O</SUP>C</FONT>孵育<FONT face="Times New Roman">1</FONT>小时,使靶细胞贴壁成一单层细胞,再向每孔加<FONT face="Times New Roman">0.1ml</FONT>效应细胞(一式三份)对照孔加<FONT face="Times New Roman">0.1ml l640</FONT>,再放<FONT face="Times New Roman">37<SUP>O</SUP>C</FONT>孵育<FONT face="Times New Roman">4</FONT>小时,取出去培养液,再用<FONT face="Times New Roman"><st1:chmetcnv UnitName="m" SourceValue="0.15" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on">0.15M</st1:chmetcnv> PH7.2 PBS</FONT>洗一次,以除去效应细胞和被杀伤而脱落下来的靶细胞,弃去<FONT face="Times New Roman">BS</FONT>液,每孔加<FONT face="Times New Roman">0.2ml0.5</FONT>醋酸乙醇(<FONT face="Times New Roman">50</FONT>%乙醇配制),溶解留下的靶细胞,摇匀,冰箱放置过夜,自每孔吸取<FONT face="Times New Roman">0.1ml</FONT>至另一<FONT face="Times New Roman">96</FONT>孔板,用酶标仪在<FONT face="Times New Roman">540nm</FONT>处测定光密度,参考波长为<FONT face="Times New Roman">630nm</FONT>。按下列公式计算溶解百分率:</FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> </FONT>对照孔<FONT face="Times New Roman">OD</FONT>值<FONT face="Times New Roman"> </FONT>- 实验孔<FONT face="Times New Roman">OD</FONT>值</FONT></P>
< ><v:line><FONT size=5></FONT></v:line><FONT size=5><FONT face="Times New Roman"> </FONT>溶解<FONT face="Times New Roman">%= </FONT>×<FONT face="Times New Roman">100% </FONT></FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> </FONT>对照孔<FONT face="Times New Roman">OD</FONT>值</FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> 2</FONT>.给药途径<FONT face="Times New Roman"> </FONT>皮肤</FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> 3</FONT>.给药次数<FONT face="Times New Roman"> </FONT>隔日<FONT face="Times New Roman">1</FONT>次,共<FONT face="Times New Roman">7</FONT>次</FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> 4</FONT>.试验对照(<FONT face="Times New Roman">1</FONT>)空白对照芝麻油(<FONT face="Times New Roman">2</FONT>)己知阳性药物对照及选择依据,本试验未设阳性药物对照组,理由同前实验。</FONT></P>
< ><FONT size=5><FONT face="Times New Roman"> 5</FONT>.试验结果见表<FONT face="Times New Roman">5</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>表<FONT face="Times New Roman">5</FONT>:镇银膏对小鼠<FONT face="Times New Roman">NK</FONT>细胞活性的影响</FONT></P>
<TABLE cellSpacing=0 cellPadding=0 border=1>
<TR >
<TD vAlign=top width=182>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>组别</FONT></P></TD>
<TD vAlign=top width=114>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>动物数</FONT></P></TD>
<TD vAlign=top width=174>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>溶解%</FONT></P></TD></TR>
<TR >
<TD vAlign=top width=182>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>空白对照组</FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>镇银膏中剂量组</FONT></P></TD>
<TD vAlign=top width=114>
<P align=left><FONT face="Times New Roman"><FONT size=5> 8</FONT></FONT></P>
<P ><FONT face="Times New Roman"><FONT size=5> 8</FONT></FONT></P></TD>
<TD vAlign=top width=174>
<P ><FONT size=5><FONT face="Times New Roman">61.27</FONT>±5<FONT face="Times New Roman">.78 </FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman">80.46</FONT>±4<FONT face="Times New Roman">.16 **</FONT></FONT></P></TD></TR></TABLE>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>与空白对照组比较<FONT face="Times New Roman"> ** P</FONT><<FONT face="Times New Roman">0. 01 <o:p></o:p></FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>结果表明,镇银膏<FONT face="Times New Roman">0.05ml/<st1:chmetcnv UnitName="g" SourceValue="10" HasSpace="False" Negative="False" NumberType="1" TCSC="0" w:st="on">10g</st1:chmetcnv></FONT>,可非常显著的增强小鼠<FONT face="Times New Roman">NK</FONT>细胞杀伤靶细胞活性<FONT face="Times New Roman">(P</FONT><<FONT face="Times New Roman">0. 01)</FONT>。</FONT></P>
<P ><FONT size=5><B >六,数据及统计学处理:</B><B ><o:p></o:p></B></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>上述试验数据均采用<FONT face="Times New Roman">T</FONT>检验</FONT></P>
<P ><FONT size=5><B >七、试验结论:</B><B ><o:p></o:p></B></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>㈠、镇银膏高、中、低剂量对小鼠和大鼠阴道上皮细胞有丝分裂均有明显的抑制作用。</FONT></P>
<P ><FONT size=5>㈡、镇银膏高、中剂量对小鼠尾鳞片颗粒细胞层的形成有明显的促进作用,但低剂量作用不明显。</FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>㈢、镇银膏高、中、低剂量可明显提高小鼠外周血<FONT face="Times New Roman">T</FONT>淋巴细胞活性,而以中剂量更为显著。</FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> </FONT>㈣、镇银膏可明显增强小鼠<FONT face="Times New Roman">NK</FONT>细胞杀伤靶细胞的活性。</FONT></P>
<P ><B ><FONT size=5>八、试验参考文献出处<o:p></o:p></FONT></B></P>
<P ><FONT size=5><FONT face="Times New Roman"> [1]</FONT>黄畋、林熙然、利用实验动物模型研究中草药内用治疗银屑病的作用,中西医结合杂志<FONT face="Times New Roman"> 1980.4</FONT>(<FONT face="Times New Roman">7</FONT>):<FONT face="Times New Roman">428</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> [2]</FONT>黄畋、林熙然<FONT face="Times New Roman"> </FONT>石蒜碱对表皮角化及细胞有丝分裂的影响<FONT face="Times New Roman"> </FONT>中华皮肤科杂志<FONT face="Times New Roman">1989</FONT>,<FONT face="Times New Roman"> 18</FONT>(<FONT face="Times New Roman">1</FONT>):<FONT face="Times New Roman">37</FONT>。</FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> [3]</FONT>苏秀玲等<FONT face="Times New Roman"> </FONT>维胺酸等维甲类衍生物对小鼠银屑病模型的实验治疗<FONT face="Times New Roman"> </FONT>中华皮肤科杂志<FONT face="Times New Roman"> 1989</FONT>,<FONT face="Times New Roman">22</FONT>(<FONT face="Times New Roman">6</FONT>):<FONT face="Times New Roman">400</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman">[4]</FONT>麻致中等<FONT face="Times New Roman"> </FONT>华蟾素治疗银屑病的临床和实验观察<FONT face="Times New Roman"> </FONT>中华皮肤科杂志<FONT face="Times New Roman"> 1989</FONT>,<FONT face="Times New Roman">22</FONT>(<FONT face="Times New Roman">4</FONT>):<FONT face="Times New Roman"> 236</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> [5]</FONT>刘贞富<FONT face="Times New Roman"> </FONT>等银屑病患者外周血<FONT face="Times New Roman">T</FONT>淋巴细胞及其亚群和天然杀伤(<FONT face="Times New Roman">NK</FONT>)细胞活性的研究<FONT face="Times New Roman"> </FONT>中华皮肤科杂志<FONT face="Times New Roman">1989</FONT>,<FONT face="Times New Roman">22</FONT>(<FONT face="Times New Roman">5</FONT>):<FONT face="Times New Roman">296</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman"> [6]</FONT>麻致中等<FONT face="Times New Roman"> </FONT>银屑病皮损直接免疫荧光及<FONT face="Times New Roman">T</FONT>细胞<FONT face="Times New Roman">ANAE</FONT>活性研究<FONT face="Times New Roman"> </FONT>中华皮肤科杂志<FONT face="Times New Roman">1989</FONT>,<FONT face="Times New Roman">22</FONT>(<FONT face="Times New Roman">4</FONT>):<FONT face="Times New Roman">225</FONT></FONT></P>
<P ><FONT size=5><FONT face="Times New Roman">[7]</FONT>喻永平等<FONT face="Times New Roman"> </FONT>用比色法测定<FONT face="Times New Roman">P0LY1</FONT>,<FONT face="Times New Roman">C</FONT>和<FONT face="Times New Roman">CP</FONT>菌苗对小鼠<FONT face="Times New Roman">NK</FONT>活性的增强作用<FONT face="Times New Roman"> </FONT>中华微生物学及免疫学杂志<FONT face="Times New Roman"> 1988</FONT>,<FONT face="Times New Roman">8</FONT>(<FONT face="Times New Roman">3</FONT>):<FONT face="Times New Roman"> 193</FONT></FONT></P>
<P ><o:p><FONT face="Times New Roman"> </FONT></o:p></P></TD></TR></TABLE>