The Effect Of Artesunate On The Expression Of Hsp47and α-SMA In Pulmonary Fibrosis In Rats

Objective:This study was to investigate the effect of artesunate onpulmonary fibrosis and the expression of Hsp47and α-SMA. The relationbetween the expression of Hsp47,α-SMA and TGFβ1/smad3was alsoobserved.This study may show more evidence for the treatment effect ofartesunate on pulmonary fibrosis and provide a new sight for anti-fibrosis in clinic probably.Method:The experiment consisted of two steps. The kinetic effect ofartesunate on BLM-induced pulmonary fibrosis in rats was observed inthe first stage. The effect of artesunate on the expression ofTGFβ1/smad3, Hsp47and α-SMA was observed in the second stage.In the first step,126rats were randomly divided into four groups.There were each24rats in control group and artesunate group, while theother two groups each consisted of39rats.①Control group: Rats were intratracheally administered with0.3ml saline and were consecutivelyintraperitoneal injected with1ml saline water per day.②Artesunategroup: Rats were intratracheally administered with0.3ml saline and wereconsecutively intraperitoneal injected with10mg/100g artesunate per day.③Model group: Rats were intratracheally administered with5mg/kgbleomycin and were consecutively intraperitoneal injected with1mlsaline water per day.④Intervention group: Rats were intratracheallyadministered with5mg/kg bleomycin and were consecutivelyintraperitoneal injected with10mg/100g artesunate per day. Six Animalsin each group were sacrificed on day7,14,21and28. Lung coefficientand death rate were calculated. The sections of the lungs were used forHE stain and Masson stain. Ashcroft score of each group were graded.Hydroxyproline content was also detected.In the following step, the rats sacrificed on day28were used in thisperiod according to the results in the first stage. The lung sections in allgroups were used for immuohistochemistry of Hsp47and α-SMA.Hsp47mRNA and Collagen Ⅰ mRNA were analyzed by RT-PCR. Theprotein of TGFβ1, smad3, Hsp47and α-SMA were also analyzed byWestern blot.Result:1(1) No rats died in control group and artesunate group. Deathrates in intervention group were lower than those in model group indifferent time.(2)Lung coefficient in model group was significant higherthan control group in all different times (P＜0.05). Lung coefficient inintervention group was not significantly different from that in modelgroup except on day28.(3)Both HE and Mason stain showed thatcompared to control group,lungs in model group were abnormal instructure and accumulated with massive collagen. Compared to model group, damage in intervention group was lighter on day21and day28.(4)Compared to control group, Ashcroft score in model group was higher inall periods(P<0.05). Compared to model group, fibrosis score inintervention group was significantly lower on day21and day28(P<0.05).(5) Compared to control group, hydroxyproline content inmodel group was higher on day21and day28(P<0.05). Compared tomodel group, hydroxyproline in intervention group was significantlylower than model group on day28(P<0.05).2(1)The expressions of Hsp47and α-SMA in immunochemistry werethe highest in model group, less in intervention group and the least incontrol group and artesunate group.(2)The expression of bothHsp47mRNA and Collagen ⅠmRNA in model group was the higher thancontrol group. All P values were smaller than0.05. The expression ofHsp47mRNA and Collagen ⅠmRNA in intervention group was lowerthan those in model group. All P values were smaller than0.05.(3)Western blot showed that the protein expression of TGFβ1, smad3,Hsp47and α-SMA in model group was obviously higher than those incontrol group. All P values were smaller than0.05. The protein levels inintervention group were all less than those in model group. All P valueswere smaller than0.05.Conclusions:1Artesunate lowers alveolitis in pulmonary fibrosis.2Artesunate attenuates collagen deposition in pulmonary fibrosis.2Artesunate plays a role in anti-fibrosis probably through downregulationof Hsp47and α-SMA.3Artesunate might decrease the expression ofHsp47and α-SMA by TGFβ1/smad3signal way inhibition.