Effects Of The Water Extract Of The Drug Shijueming On Endoplasmic Reticulum Stress In Lens Epithelial Cells Of Rats With D-galactose Cataract

The topic was aimed to investigate the mechanism by which D-galactose induced endoplasmic reticulum stress in lens epithelial cells of rats with cataract, and study effects of the water extract of the drug shijueming on lens of rats with D-galactose cataract.Part one Preparation.Purpose D-galactose cataract models were established.Methods1One hurdred healthy SD rats(40days,200±10g) were randomly divided into four groups(blank group,control group, baineiting eyedrops group, shijueming eyedrops group), both eyes were treated drugs(200lens).2D-galactose cataract models were established.3Blank group and control group were treated with physiological saline eyedrops. Baineiting eyedrops group was treated with Baineiting eyedrops. Shijueming eyedrops group was treated with0.25%Shijueming eyedrops(three times each day, one drop each time).4On15th day.One hundred lens of SD rats were extracted by strict aseptic manipulation after rats executed,then lens were keep in-80℃fridge.Results To observe the lens of rats after mydriasis on15th day, the stage of cortical opacities in control group achieved Ⅱ.refering to standard stage of cortical opacities invented by Qi Mingxing and Huang Xiurong. Cortical opacities of lens in shijueming eyedrops group developed slower compared with control group. The stage of cortical opacities in baineiting eyedrops group and shijueming eyedrops group achieved I.The lens of blank group were transparent.Conclusion The water extract of the drug shijueming can delay cataracts of rats.Part two The levels of CHOP,GRP78and XBP-lmRNA were measured by immunohistochemistry, Western blot and RT-PCR methods.Purpose To investigate further effects of the water extract of the drug shijueming on lens epithelial cells of rats with D-galactose cataract. Methods The levels of CHOP,GRP78and XBP-1mRNA by immunohistochemistry, Western blot and RT-PCR methods of four group were measured.Results1CHOP was measured by immunohistochemistry methodTo observe the slice of lens by light microscope (Fig1).The postive rate of cell in blank group was5.74+3.20%, the postive rate-of cell in control group was92.62+2.32%, the postive rate of cell in baineiting eyedrops group was45.72+3.51%, the postive rate of cell in shijueming eyedrops group was44.77+4.54%(Tab1), The postive rate of cell in shijueming eyedrops group was lower than in control group(P<0.05). The postive rate of cell in shijueming eyedrops group was was similar to baineiting eyedrops group(P>0.05). The postive rate of cell in control group was higher than in blank group obviously (P<0.01).2GRP78was measured by Western blot methodThe expression in blank group was0.013±0.21μg/ml, the expression in control group was0.556±0.16μg/ml, the expression in baineiting eyedrops group was0.321±0.26μg/ml, the expression in shijueming eyedrops group was0.212±0.15μg/ml (Tab1) The expression in shijueming eyedrops group was lower than in control group (P<0.05). The expression in shijueming eyedrops group was was similar to baineiting eyedrops group(P>0.05). The expression in blank were lower obviously compared with control group (P<0.01).3XBP-1mRNA was measured by RT-PCR methodThe expression in control group was9.2±0.04, the expression in baineiting eyedrops group was6.5±0.02. the expression in shijueming eyedrops group was2.4±0.03, the expression in blank group was1.0±0.02(Tab1). The expression in shijueming eyedrops group was lower than in control group (P<0.05). The expression in shijueming eyedrops group was lower than in baineiting eyedrops group(P<0.05). The expression in blank were lower obviously compared with control group (P<0.01).Conclusion The water extract of the drug shijueming can decrease endoplasmic reticulum stress in lens epithelial cells of rats with D-galactose cataract and protect lens. It also can delay the development of cataract.